Asian Journal of Transfusion Science
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Year : 2016  |  Volume : 10  |  Issue : 3  |  Page : 8-33
40 th ISBTI Annual Conference, TRANSCON 2015, New Delhi

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Date of Web Publication19-May-2016

How to cite this article:
. 40 th ISBTI Annual Conference, TRANSCON 2015, New Delhi. Asian J Transfus Sci 2016;10, Suppl S1:8-33

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. 40 th ISBTI Annual Conference, TRANSCON 2015, New Delhi. Asian J Transfus Sci [serial online] 2016 [cited 2022 May 23];10, Suppl S1:8-33. Available from:


Frequency and pattern of transfusion in obstetrics patients

Vibha Singh, Anil Gupta, Neetu Singh, Deepti Agrawal, AK Verma, Ajay Sagar

Department of Transfusion Medicine, ESI Hospital, New Delhi, India

Background: Patients of Obstetrics and Gynaecology department require frequent blood transfusion due to anemia of pregnancy itself or associated ailment.

Aim: This study was done to evaluate blood transfusion pattern, frequencies and nature of demand amongst patients admitted in Obstetrics ward.

Methods: All patients admitted during first quarter of 2015 in obstetrics wards of ESI hospital, Delhi, were evaluated for their transfusion related demands and various diseases and conditions influencing the demand; including age, severity of anemia, number of request order received, no of units transfused, Cross-match Transfusion ratio͹ T ratio) and co morbidity if any were noted.

Results: A total number of 850(n) transfusion request were received from Obstetrics department, out of which 63% were for multi-gravida (535), 34.8% were primi gravida (n = 296) and 2.2% were grand multi gravida (n = 19). It was found that 52% patients with hemoglobin between 7.1 to 10.9 g/dl needed less blood transfusion with a C T ratio of 8.8:1. However patient with severe anemia (Hb 7 or less) constituted 14% of total Obstetrics population; and required greater number of transfusion with a higher C T ratio (1.2:1).This group of obstetrics Patients reflects higher vulnerability and poor hemodynamic status during pregnancy. Anemia is one cause which precipitates pre-term labour. Nearly 50% of pregnant women with pre-term labour received blood transfusion.

Pregnancy Induced Hypertension (PIH) was an important condition and was associated with increased blood transfusion demand in obstetrics patients. About 44% patients (11/25) in this group received blood transfusion due to severe anemia.

IUGR and IUD were other important conditions which necessitate blood transfusion. IUGR and IUD were precipitated by sever anemia leading to septicemia or impending labour. In few of cases at our hospital eg. placental abnormality, ectopic pregnancy, incomplete abortion and heart disease were also responsible for increase requirement of blood transfusion.

Conclusion: Although almost each patient admitted in Obstetrics department was asked for blood transfusion due to fear of impending bleeding during labour but only 26.7% received blood transfusion. Transfusion was higher in patient with Hb less than 7.0 g% and in patients with associated maternal and fetal complications. The CT ratio is very high in patient with mild to moderate anemia which can be reduced to the universally acceptable level by way of senstization of obstetrician through frequent CMEs, seminars and involving them in our national conference. It is to emphasized that patients with severe anemia and associated maternal/fetal ailment need to be supervised closely for better and effective blood transfusions.

Safety and efficacy of small volume plasmapheresis in children: Experience in a tertiaty neurosciences centre

Sundar Periyavan, Sukanya Baruah, Sangeetha K Sheshagiri, N Shivanna, J Vivekanand, P Shnathi

Background: Therapeutic Plasma Exchange is the standard treatment in various immune mediated neurological disorders. The procedure is done by automated or manual methods. We have done manual method in children with less body weight, removing small volume of plasma (small volume plasmapheresis).

Aim: The aim of the study was to find the safety and efficacy of small volume plasma exchange.

Materials and Methods: The study was performed over a period of one year from 1 May 2014 to 30 April 2015, at Transfusion Medicine Centre, National Institute of Mental Health and Neurosciences (NIMHANS), Bengaluru, South India. The study included 30 children in the age group of 3 to 17 years. There were 24 cases (80%) of Guillain-Barre Syndrome and 6 (20%) cases of autoimmune encephalitis. Of these 15 were males and 15 females. The body weight of all the patients was below 35 kgs, the minimum being 10 Kgs and maximum 35 Kgs.

Whole blood was collected into a double bag, centrifuged at 5000 g for 10 minutes at four degree centigrade. All the plasma was expressed under laminar flow bench into a satellite bag, the tubing was sealed and the satellite bag was separated. The plasma was discarded as per standard protocol. Requisite amount of normal saline was added to the packed cells with sterile infusion set in ultraviolet hood and sent to the ward for infusion to the patient. One unit FFP was used as replacement for protein loss, on alternate days. A total of 247 procedures were performed. The maximum was 20 exchanges in two patients (0.6%) of autoimmune encephalitis and the minimum was three exchanges in three (0.9%) patients of GBS. The rest of the patients underwent five to ten procedures.

Results: No procedure related complications occurred in any of the patients. Six (18.1%) patients had allergic reactions to FFP infusion in the form of rash and itching. The reactions were mild to moderate in nature and subsided with antihistaminic agents. All the children with GBS and autoimmune encephalitis showed signs of arrest of the progression of the disease and later showed clinical improvement during the stay at hospital. Long term follow up could not be done.

Conclusion: From our experience TPE can be considered to be a safe and effective procedure in children with neurological disorders with benefits outweighing risks. This procedure is can be done hospitals attached blood banks having blood component facility but no apheresis machine, this will avoid huge expenditure involved in automated plasmapheresis or IVIg.

Prevalence of HIV, HCV, HBV infection in blood donors detected by nucleic acid testing: An Indian

Tulika Chandra, Devisha Agarwal

Department of Transfusion Medicine and Blood Bank, King Georg's Medical University, Lucknow, Uttar Pradesh, India

Background: Nucleic acid amplification testing (NAT) and is based upon the technique of direct amplification and detection of viral nucleic acids rather than antibody production by the immune system of the infected person. NAT is thus able to detect viruses during the 'window period' or the time between donor exposure to the virus and the appearance of antibodies. This allows for earlier detection of infection and further decreases the possibility of transmission via transfusion.

Aim: To compare the rate of detection between routine ELISA tests and NAT testing.

Methods: The samples of the donors collected in blood bank after careful history and examination were tested by ELISA. The ELISA negative samples were tested for NAT and the data was analysed.

Results: Out of the 35,722 samples, 700 (1.95%) were reactive by ELISA. Discriminant assays found 40 (0.11%) to be reactive for HIV, 60 (0.16%) to be reactive for HCV and 600 (1.67%) for HBV. Apart from these reactive samples NAT was carried out on the non-reactive samples. Amongst them a total of 158 (0.44%) were reactive for NAT. On discriminatory assays 2 (0.005%) samples were reactive for HIV-1 and 2, 46 (0.12%) samples were reactive for HCV and 108 (0.30%) for HBV.

Conclusion: NAT could detect HIV, HBV and HCV cases in blood donor samples that were undetected by serological tests. Third generation ELISA is the mandatory test in India. Our study confirms the utility of NAT in Indian donors.

Study on such a huge population sample further questions the safety of blood supply by regular screening method and stresses on the introduction of NAT as a preferred test in most of the blood banks.

Key words: ELISA, HBV, HCV, HIV, NAT

Blood usage in standalone blood bank: A retrospective study

Sheenam Sood, Mahadeo Mane

Department of Immunohaematology and Blood Transfusion, Dr. D. Y. Patil Hospital and Research Centre, Kolhapur, Maharashtra, India

Background: Blood transfusion constitutes an important part of various treatment protocols. Indications for blood use must be clear in the mind of ordering clinicians, to avoid its misuse and also to avoid unnecessary exposure of the patient to donor blood antigens, adverse reactions and transfusion transmissible diseases.

Aim: To analyse the requisition of blood and blood components in a standalone blood bank.

Methods: A retrospective study of blood transfusion requisitions in a standalone blood bank at Kolhapur, Maharashtra, India, for a period of 1year, from January 2014 to December 2014, was carried out. The analysis of blood and its component requisitions in all patients from different hospitals was reviewed regarding diagnosis, indication for transfusion, number of units requested and the speciality prescribing it.

Results: Total number of blood component requisitions were 13,485. Out of which 196 requests were for whole blood, 7457 requests were for packed red cells, 4230 were for FFP and 1602 for platelet concentrate. Packed red blood cells were the most utilized product. Supply of blood was maximum to the medicine wards. The most common indication for whole blood, packed red cells and fresh frozen plasma requests was anaemia. Whereas the most common indication for platelet concentrate requests was thrombocytopenia.

Conclusion: Clinical audits help to reduce inappropriate use of blood and blood components and also avoids blood transfusion reaction. Periodic review of blood component usage is important to assess the blood utilization pattern in any geographical area.

Hepatitis E associated autoimmune hemolytic anemia

Sheenam Sood, Mahadeo Mane

Department of Immunohaematology and Blood Transfusion, Dr. D. Y. Patil Hospital and Research Centre, Kolhapur, Maharashtra, India

Autoimmune hemolytic anemia is a rare disease characterized by hemolysis which leading to various life-threatening conditions that requires blood transfusion. In this disease autoantibodies are directed against red blood cells which causes their premature destruction by macrophages from the reticuloendothelial system. Warm antibody type autoimmune hemolytic anemia is the most common autoimmune hemolytic anemia (AIHA). Secondary AIHA due to hepatitis viruses like HAV, HBV, HCV are documented, however AIHA due to Hepatitis E is infrequent. Here we report a case of Secondary AIHA associated with HEV. In our case Coombs test (DAT) and antibody against HEV were positive.

Methods: A retrospective follow up of a Patient.

Conclusion: Reported case is of secondary AIHA associated with Hepatitis E.

Autologous blood transfusion as a life saving measure for a trauma patient with fracture femur and drug induced hemolytic anemia: A case report

Sumit Vishwakarma, Sumit Vishwakarma, Vedanand Arya, Arulselvi Subramanian, Vivek Trikha, Rajesh Thukral, Poonam Kosic, Kabita Chatterjee

Departments of Laboratory Medicine and Blood Bank and Orthopedics, Jai Prakash Narayan Apex Trauma Centre, AIIMS, New Delhi, India

Background: The direct antiglobulin test is used to determine whether red blood cells (RBCs) have been coated in vivo with immunoglobulin, complement, or both. The indirect antiglobulin test is used to determine the presence of antibody in the serum or plasma. There are many causes of a positive direct antiglobulin test. Depending on the technique and the reagents used, a positive direct antiglobulin test has been reported in 1:1000 to 1:14,000 blood donors and 1%-15% of hospital patients. Most blood donors with positive direct antiglobulin test results appear healthy, and most show no overt signs of hemolytic anemia. Drugs may cause a positive direct antiglobulin test result and/or immune-mediated hemolysis with an incidence of approximately 1 in 1 million.

Aims: To highlight the importance of following strict transfusion protocols and management insight in an Apex trauma center setting in a coombs test positive patient.

Case Report: 36 year old male alleged h/o RTA with injury to left thigh on April 2015. H/o- ankylosing spondylitis since 20 years on medication (indomethacin and methotrexate). X-ray- comminuted intra-articular fracture of the distal femur. Demand for blood was initiated but cross match of blood was incompatible with multiple units and patient was direct and indirect coombs test positive. Later on two units of autologous blood units were harvested and transfused. Patient was operated for his fracture and the postoperative period was uneventful.

Discussion and Review of Literature: Penicillin and some other drugs (the antigen) may bind loosely to RBCs in vivo, thus becoming immunogenic and stimulating the production of antibodies. After antibody production is initiated, immune complexes form (consisting of antibody and drug) and these complexes in turn bind non-specifically to other RBCs and ultimately lead to the activation of complement system. This "immune complex" mechanism usually generates a severe intravascular reaction, with a positive DAT. It is true that hemolytic transfusion reactions are expected to occur when incompatibility is due to clinically important alloantibodies. Experience indicates that when incompatibility is only due to the presence of a RBC autoantibody, the survival of transfused RBCs is generally about as good as that of the patient's own RBCs, and transfusion can be expected to cause significant temporary benefit. In the absence of infection or tumor the use of prebanked and Intraoperative autologous transfusion is recommended in orthopedic procedures in which homologous transfusion may be needed.

Take Home Message: If patient has a broadly reactive auto/alloantibody, the transfusion service is likely to find that many units of RBCs are incompatible, adding an uncertainty to the risk-benefit ratio of transfusion. Pre-operative autologous blood donation (PABD) aims to provide a supply of safe blood for patients undergoing surgery who might need a blood transfusion before scheduled elective surgery.

Prevalence and trends of transfusion transmitted infections among blood donors of blood bank attached to Government Hospital of South Gujarat

Kruti Raja, Jitendra Patel, Amrish Pandya, Gopi Dobariya, Snehal Patel, Sangita Wadhwani

Department of IHBT, Government Medical College, New Civil Hospital, Surat, Gujarat, India

Background: Though the blood transfusion is life saving, it is never risk free. Blood transfusion is having potential risk for transfusion transmitted infections (TTIs). The risk of TTIs is estimated to be 1 in 6, 77,000 units for HIV, 1 in 1, 03,000 for hepatitis C virus and 1 in 63,000 for hepatitis B virus. Magnitude of TTIs varies from country to country.

Objective: The objective of present study is to assess the prevalence and trend of TTIs among blood donors in the blood bank attached to tertiary level government hospital.

Materials and Methods: A retrospective review of donors' record covering the period from January 2011 to May 2015 was done in the present study. All samples were screened by 3 rd generation ELISA methods for HIV 1 & 2, HBsAg and HCV. HIV 4 th generation ELISA test was also done during the year 2014-2015. Screening for Syphilis was done by rapid (RPR and TPHA)/ELISA method and Malaria screening was done by either peripheral smear or rapid tests.

Results: Out of 35106 donors, over all sero prevalence was 484 (1.38%) donors. Out of 484 sero reactive donors, 32 (0.09%) were reactive for HIV, 345 (0.98 %) for HBsAg, 32 (0.09 %) for HCV, 70 (0.20 %) for Syphilis and 05 (0.01 %) were reactive for Malaria.

Conclusion: With the implementation of strict donor criteria and use of sensitive screening test it may be possible to reduce the incidence of TTIs in Indian scenario. After the year of 2012, there was a decrease in sero prevalence of HIV among blood donors of the blood bank (0.2 % in the year 2012 vs. 0.04 % in the year 2014).

Significance of vitros antihbcore assay in detecting occult HBV infection in donor samples

Anju Verma, Sushila Yadav, Anju Verma

Rotary Blood Bank, New Delhi, India

Background: Hepatitis B core antibody (Anti HBc) is the first antibody to appear following acute Hepatitis B infection and persist in high levels following resolution of infection and in chronically infected patients. A small number of Hepatitis B virus (HBV) carriers appear to circulate Hepatitis B surface antigen (HBsAg) at undetectable levels, and anti-HBc may be the only serologic marker detectable in blood in these individuals. During the "window period," when antigenemia with HBsAg has resolved and anti HBsAg (anti-HBs) has not yet developed, HBc IgM antibody may be the only marker present. As the appearance of anti-HBsAg may be delayed after HBsAg clearance, anti-HBc is sometimes the only serological marker for HBV infection and potentially infectious blood. In certain instances, isolated anti HBc may be observed. Isolated anti-HBc is defined as the presence of anti-HBc in the absence of detectable HBsAg and hepatitis B surface antibody (anti-HBs). Many patients with occult HBV will have anti-HBc as the only serologic marker to suggest HBV infection.

Occult HBV infection is characterized by the presence of HBV DNA in serum and/or in the liver of patients negative for hepatitis B surface antigen (HBsAg) but may be positive for anti HBc antibody. Occult infection may impact in several different clinical contexts including the risk of HBV Transmission with transfusion or transplantation, and endogenous viral reactivation. Serological assay for the long-lasting antibody response to the highly immunogenic HBV core Antigen (anti-HBc) represents a qualified candidate as a surrogate for DNA amplification, and for increasing overall sensitivity when assessing the risk of occult hepatitis in peripheral blood. The risk of occult hepatitis associated with anti-HBc seropositivity has been demonstrated extensively.

Objective: To study the significance of anti HBc antibody screening in healthy donor samples to detect the Hepatitis B Occult infection and to evaluate the 'isolated HB core antibody alone' reactive samples which showed non-reactive for HBsAg and HBV DNA.

Protocol: Rotary Blood Bank, New Delhi issues approximately 35000 units of blood or blood components per year to the patients in need. In Rotary blood bank, highly sensitive Enhanced Chemiluminescence technology is being used for infectious disease screening viz., Anti HIV, Anti HCV, HBsAg and HBc antibody in all blood donor samples for which Vitros 3600 Immunodiagnostics system is used to enhance the safety of the blood for transfusion. Further, to detect any sero-negative 'window period' infected donor samples, all the samples were subjected to NAT testing using Chiron's multiplex Procleix Ultrio Assay.

In this study a total of 14220 healthy donor blood samples were screened for all infectious disease markers using VITROS 3600 Immunodiagnostics system followed by NAT testing. All the donor samples, which showed 'isolated anti HBc alone' reactive in VITROS anti HBc competitive immunoassay were subjected to Biorad Anti HBc ELISA assay which is based on indirect immunoassay, to rule out any false reactivity in VITROS anti HBc antibody assay. The donor samples which showed 'Non-reactive' in Biorad Anti HBc ELISA assay were excluded from the study. All the donor samples, with confirmed 'isolated anti HBc alone' reactive in both anti HBc assays were subjected to VITROS anti HBs assay to quantify the anti HBsAg antibody levels.

A total number of 25 randomly selected 'Isolated HBc antibody alone reactive' samples with NAT Negative, HBsAg Negative and Anti HBs negative (<10 mIU/mL) donor samples were subjected to high sensitive HBV DNA PCR (HBV Viral Load) Quantitative assay with Cobas Taqman kit. This helped to identify if there is any low level of HBV DNA present in these samples of 'isolated Anti HBc alone reactive' donors, which were missed by Chiron's multiplex Procleix Ultrio Assay.

Results: Out of 14220 Donors screened, 813 (5.7%) were found reactive for HBc antibody in VITROS HBc antibody assay. All the 813 HBc antibody reactive samples were re verified in Biorad Anti HBc assay. Out of 813 Anti HBc initial reactive Donors screened, 493 samples showed repeat reactive in Bio-Rad Anti HBc ELISA technique. All the 493 Donors were quantified for Anti HBs level using VITROS anti HBs assay and based on the antibody level, they were consolidated in 3 groups [Table 1].

Out of 85 samples, which were 'isolated HBc antibody alone' reactive with anti HBs (<10 mIU/mL) 25 samples were randomly selected and subjected to HBV DNA PCR (HBV Viral Load) Quantitative with Cobas Taqman kit. Out of 25 samples tested, 02 samples (8%) showed the presence of HBV DNA which were missed by the Chiron's multiplex Procleix Ultrio Assay. These samples with Occult HBV infection with very low level of HBV viral load showed the presence of 'isolated HBc antibody', which is the only serological marker to suggest HBV infection [Table 2].


This Study supports that Donors with occult HBV infection, who lacks detectable HBsAg but having exposure to HBV infection was indicated by a Positive Anti HBc antibody with or without detectable HBV DNA in circulation, are a potential source of HBV infection

The VITROS Anti-HBc Assay has the sensitivity of picking up the Occult HBV Donors which were missed by HBsAg assay and NAT test

Anti HB core antibody screening helps in detecting the Occult HBV infection and enhances the safety of blood for transfusion.

Frequency and distribution of abo and Rh blood groups among blood donors in tertiary care hospital of South Gujarat

Gopi Dobariya

Department of IHBT, Government Medical College, Surat, Gujarat, India

Background and Objectives: Since it was discovered by Karl Landsteiner, the ABO blood group system is the most important blood group system in Transfusion Medicine. The blood group systems are also very important in population genetic studies, researching population migration patterns as well as resolving certain medico-legal issues, particularly disputed parentage. This retrospective study was carried out with an objective to provide data regarding frequency and distribution of ABO and Rh blood groups among blood donors in South Gujarat.

Materials and Methods: Data of 32862 blood donors were retrospectively collected and analyzed regarding ABO and Rh blood groups from May-2011 to May-2015 and reported in simple numbers and percentage. Blood group of the blood donors was determined by forward & reverse methods with the help of commercially available standard monoclonal antisera by test tube & column agglutination techniques in required cases.

Results: The most common blood group among donors was B (34.37%) followed by O (32.28%), A (24.38%), while the least prevalent blood group was AB (8.96%). Rh positivity among donors was 95.01%. Rests were Rh Negative (4.99%).

Saline washed red cell concentrate: A preferable component for multi transfused patients

Tanvi G Patel, Rinku V Shukla, Narendra V Vasavada

Surat Raktadan Kendra and Research Centre, Surat, Gujarat, India

Background: Transfusion reactions like allergic reactions and febrile reactions may occur due to foreign plasma proteins and leukoaglutinins present in packed red cells. Saline washed red blood cells (RBC) are transfused in multi transfused patients of thalassmia, sickle cell anemia, having recurrent febrile reaction, etc to minimize these reactions.

Aim: To estimate leukoreduction, red cell loss and plasma removal in saline washed RBC and assess its quality.

Methods: In this study, we included 170 saline wash RBC. Volume was measured on electronic weighing scale (LABTOP) before (pre) and after (post) saline wash. Two samples of 2 ml were taken from blood unit by stripping the tubing in laminar air flow. One sample was used for measurement of hematological parameters and the other tube was centrifuged for 3000 rpm for 5 minutes. Supernatant plasma was separated and used for protein estimation using spectrophotometer. The blood bag was washed twice with sterile normal saline (Baxter) according to standard operating procedure and the 1 ml supernatant was collected for post sample protein estimation. The post sample was collected from bag for hematological analysis .RBC loss, leukoreduction and amount of plasma removed were calculated.

Results: The volume of pre saline wash unit was 306.14 ΁ 31.19 ml and post was 291.26 ΁ 34.18 ml. The RBC count of pre sample was 2.32 ΁ 0.35 Χ 1012/unit and post was 2.02 ΁ 0.32 Χ 1012/unit. The RBC loss in saline wash RCC was 12.46 ΁ 6.34%. The WBC count of pre sample was 3.06 ΁ 1.27 Χ 109/unit and post was 0.24 ΁ 0.31 Χ 109/unit. Leukoreduction was 92.10 ΁ 7.33%. The total protein of pre sample was 6.60 ΁ 0.72 gm/dl and post was 0.42 ΁ 0.09gm/dl.

Conclusion: In saline wash procedure we get almost log 1 leukoreduction and acceptable RBC loss according to standards. Removal of 93.52% plasma can prevent transfusion reaction. Saline wash RCC is cost effective and preferable Leucoreduced product for multitransfused patient if used within 24 h.

Analysis of indications and pattern of usage of fresh frozen plasma

Amruta Khade, Mahadeo Mane

Department of Immunohaematology and Blood Transfusion, Dr. D. Y. Patil Hospital and Research Centre, Kolhapur, Maharashtra, India

Background: The appropriate use of blood and blood components means, the transfusion of safe blood components, only to treat a condition leading to significant morbidity or mortality, which cannot be prevented or managed effectively by other means. Fresh frozen plasma contains near normal levels of many plasma proteins, including procoagulant and inhibitory components of the coagulation cascades, acute phase proteins, immunoglobulins and albumin. This study was conducted to review the practice of Fresh Frozen Plasma usage for transfusion, based on the coagulation profile, requested by various departments in the D. Y. Patil Hospital, Kolhapur.

Aim: To evaluate indications and appropriateness of Fresh Frozen Plasma transfusion according to guidelines of College of American Pathologists.

Methods: Prospective study was done at Blood Bank of D. Y. Patil Hospital, Kolhapur, over a period of 5 months starting from 1st of January 2015 to 31 st May 2015. The following data was collected whenever a request for Fresh Frozen Plasma was received. 1) Clinical profile 2) diagnosis of the patient 3) indications. Guidelines of College of American Pathology were followed. ABO & Rh compatible Fresh Frozen Plasma were issued to the respective department. Post transfusion Prothrombin Time was done within one hour after completion of transfusion.

Results: Total 698 units of FFP were issued during study period for 155 transfusion episodes. Bleeding, Liver disease, DIC, Coagulopathy, Surgery were common indications, while FFP was also utilized in patients with Anemia, GB Syndrome and Post operative state. Analysis highlighted that only 453 units (65%) were used for appropriate indications. The effectiveness as judged by Prothrombin Time.

Conclusion: In long term practice, limiting the use of blood components only for appropriate indications will result in patients receiving optimal treatment with the lowest risk of side effects and transmission of infectious agents. This practice will prevent wastage of FFP avoid shortage in times of crisis and minimize the treatment cost.

Hepatitis B vaccination in blood bank workers

Amruta Khade, Mahadeo Mane

Department of Immunohaematology and Blood Transfusion, Dr. D. Y. Patil Hospital and Research Centre, Kolhapur, Maharashtra, India

Background: Blood Bank workers are not aware about the pathogenic effects of Hepatitis B virus which can be transmitted by handling blood and blood components. This Group of workers is a high risk group. They are unaware about morbidity and mortality caused by Hepatitis B virus. Knowledge about exposure to Hepatitis B virus is not known to most of the workers. This study was conducted to know the awareness about Hepatitis B vaccination.

Aim: To know the need, awareness and importance of Hepatitis B vaccination IN Blood Bank workers.

Materials and Methods: Data collected from 5 Blood Bank staff working in Blood Banks located in Kolhapur city, regarding Hepatitis B vaccination. Blood Bank staff can be categorized into 2 main categories.

Those who look after the administrative work of Blood Bank (e.g., PRO, Cashier, Clerk, Computer Operator)

Those who handle Blood, Blood components and Blood donors (e.g. Tecnicians, Reception Staff, BTO, Call Boys, Material Washing Staff, sweeper).

Workers in category 2 were interviewed and data prepared, for that questionnaire. Was prepared and submitted to respective workers. From the answers received data was prepared. Total 100 workers interviewed in category 2 were considered.

Observation: From the data received we categorized workers in 3 Types as 1) fully vaccinated. 2) Incompletely vaccinated. 3) Not vaccinated at all. Of which fully vaccinated were 14%.

Conclusion: Most of workers have not taken Hepatitis B vaccination. Proper counseling and creating awareness about Hepatitis B virus may motivate blood bank workers to undertake full vaccination schedule against Hepatitis B virus.

TTI reactivity rate in repeat and first time blood donors: A 5 year trend analysis

Sandeep Choudhari, Shashikant Patil, Nandkishor Tated, Atul Jain, Ratnakar Kasodkar

Background: Introduction of early detection tests such as NAT Testing facilities increased blood safety to a larger extent. However, there are still NAT yield or breakthrough infections of TTIs. Voluntary non-remunerated repeat blood donors are perceived to be safer than replacement and first time blood donors.

Aim: To analyse the trend of TTI Reactivity rate in Repeat & First time Blood Donors at our centre.

Materials and Methods: The screening results of 1,20,067 blood donations between January 2010 and December 2014 were analyzed for prevalence of hepatitis B virus (HBV), hepatitis C virus (HCV) & human immunodeficiency virus (HIV) infections. The trend of TTI positivity among repeat and first time donors was studied.

Results: Increase in our Repeat Donation rate has been 36.88% in these 5 years [53.33% (13363/25057) in 2010 to 73% (15273/20921) in 2014]. Overall TTI reactivity was 1.24% (1498/1,20,067) amongst which TTI reactivity in First time donors was 2.78% (1242/44621) and in repeat donors was 0.33% (256/75442). First time donor TTI reactivity rate was 9 times greater than Repeat donor TTI reactivity rate. Over the period of 5 years overall TTI reactivity of our donors decreased by 11.6% (1.37% in 2010 to 1.21% in 2014). Increasing trend of TTI reactivity was seen in repeat donors over the period of 5 years [0.1 % (16/13363) in 2010 to 0.48 % (74/15273) in 2014].

Conclusion: Repeat donation has increased over the period of 5 years. A decreasing trend of overall TTI reactivity was seen over the period. Sero-reactivity rate in repeat donors was lesser than that in first time donors. However repeat voluntary blood donors showed marginally increasing trend of TTI reactivity which is a matter of concern. Therefore it would be advisable to direct more focus & attention on implementation of proper donor counselling and a proper system to prevent the TTI reactive repeat donors from donating again and again.

Seroprevalence of anti-hepatitis C antibodies among healthy blood donors at a Tertiary Care Teaching Hospital in Punjab

Neha Syal, Neetu Kukar, RN Maharishi, Arunpreet Kaur, Harkiran Arora

Department of Transfusion Medicine, Guru Gobind Singh Medical College and Hospital, Faridkot

Background: Blood transfusion has an important role in the modern practice of medicine. However there must be a sound reason for transfusion in any patient because transfusion exposes the patient to several types of risks including the possibility of transmission of infectious agents like HIV, HBsAg, HCV, Malaria and Syphilis.

Aims: This study was undertaken to assess the seroprevalence of HCV in voluntary and replacement blood donors at a Tertiary Care Teaching Hospital in Punjab and to evaluate the trends over the years.

Methods: The present retrospective study was undertaken on healthy blood donors in the department of Immunohaematology and Blood Transfusion at Guru Gobind Singh Medical College and Hospital, Faridkot. The study was conducted to know the seroprevalence of HCV in healthy blood donors over a period of three years from Jan 2012 to Dec 2014. Blood samples collected in sterile plain vials were centrifuged to separate serum. The sera were then tested for HCV antibodies using 3 rd generation ELISA kits.

Results: A total of 31147 donors were screened for HCV seropositivity in three years period. Seropositivity was 2.25% in 2012, 2.3% in 2013 and 2.6% in 2014. Trends over the period are almost the same and HCV seropositivity ranged from 2.25 to 2.6%. Seropositivity in replacement donors was 3.29% in 2012, 2.9% in 2013 and 4.08% in 2014 which is very high as compared to that of voluntary donors having seropositivity of 1.85%, 2.06% and 2.23% respectively which is highly significant (p < 0.001).

Summary: Indian studies indicate the seroprevalence of HCV between 0.4% to 1.09% among blood donors. Garg et al. reported HCV prevalence of 0.28% in blood donors of Western India, Thakral et al. (2005) reported a rate of 0.44% at PGI, Chandigarh and a rate of 0.54% was reported by A. Kurl et al. (2003) at Ludhiana. The high seropositivity of HCV to the tune of more than 2% among healthy blood donors in Punjab calls for an aggressive community education programme regarding the risk factors of HCV and screening of family members of all seropositive cases. Also this highlights the importance of concentrating on some alternative mode of transmission of this silent killer as the patients of Hepatitis C presenting to the clinicians are just tip of the iceberg. Higher seropositivity in replacement donors indicate the importance of promoting voluntary blood donors as they are the cornerstone of safe and adequate supply of blood.

Acute transfusion reactions: Blood safety monitored by hemovigilance in a tertiary care centre

Sangeetha Gayam, K Hitesh Kumar, S Venkat, Sikinder Hayath

Department of Transfusion Medicine, KIMS, Nalgonda, Telangana, India

Background: Scope of different hemovigilance systems varies due to differences in spectrum of reporting all adverse events as only serious adverse reactions. [1]

ISBT has laid down criteria for severity and imputability of transfusion reactions. [2]

Aim: To evaluate transfusion reactions in the recepients of blood admitted to KIMS Hospital, Narketpally, Nalgonda district, Telangana state.

Methods: Transfusion reaction traceability document is adopted to assess the transfusion reactions in the year 2014. Imputability level criteria issued by IPC (Indian Pharmacopoeia Commission) are followed.

Results: 7 cases are documented with transfusion reaction. The duration of commencement of reaction varied from less than 10 minutes to 3 hours 30 minutes. The commonest reaction observed is burning sensationat venepuncture site i,e in 3 cases. Chills, fever, pain, sweating are seen one in each of the remaining 4 cases. The commonest imputability level noticed is "definite" type. All transfusion reactions are documented in blood bags issued which are 20 to 33 days old.

Summary: 7 cases of transfusion reactions are documented at KIMS, Narketpally in the year 2014, which are due to blood issued of 20 to 33 days old. The commonest transfusion reaction noticed is burning sensation at venepuncture site (4 cases). The "definitive" imputability level is noticed in 4 cases but "possible" imputability level is noticed in 1 case and "probable" imputability level in 2 cases.

Hence other causes may be the contributory causes in 3 cases.

Near expiry date issue blood need not be taken as the only criteria for transfusion reaction, as Direct Coomb's Test is negative in all the seven cases in the post transfusion blood samples of the recepients.


1. de Vries RR, Faber JC, Strengers PF; Board of the International Haemovigilance Network. Haemovigilance: an effective tool for improving transfusion practice. Vox Sang 2011;100:60-7.

2. Sainsby D, Faber JC, Jorgensen J. Overview of hemovigilance. In: Simon TL, Solheim BG, Straus RG, Snyder EL, Stowell CP, Petrides M, editors. Rossi's Principles of Transfusion Medicine. 4 th ed. West Sussex, UK: Blackwell Publishing; 2009. p. 694.

Rantes: A chemokine in different types of stored platelet concentrates

Rinku Vishal Shukla, Tanvi G Patel 1 , Snehalata C Gupte 1

Surat Raktadan Kendra and Research Centre, 1 Surat Raktadan Kendra and Research Centre (NABH Accredited Regional Blood Transfusion Centre), Surat, Gujarat, India

Background: Transfusion of platelets may be responsible in many non hemolytic transfusion reactions. They contain several mediators that belong to a family of pro inflammatory cytokine-chemokine that are stored in the organelles. Regulated upon activation, normal T cells expressed and secreted (RANTES) is released and accumulated in stored platelet concentrate and is mainly involved in allergic reactions.

Aims: To measure the level of RANTES in different types of platelet concentrate (PC) at different time interval of storage.

Methods: Fifteen PCs were prepared by platelet rich plasma (PRP) and buffy coat (BC) method each. Forty-two SDP were prepared using three cell separators Cobe Spectra, Trima and Amicus. Filtered PCs were prepared using 10 lab side filters and 6 bed side filters. The supernatants of PRP and BC PCs were collected aseptically after one hour, 18 hrs, 65 hrs and 112 hrs of preparation. Platelet count was done using hematology analyzer. SDP samples were taken on 0 day, 3 rd day and 5 th day. In filtered PC pre and post filtration samples were taken for measuring platelet count. All platelet concentrates were stored for five days at 22ΊC in platelet agitator. The supernatant from all samples was frozen in aliquots at -56oC for measurement of RANTES concentration using ELISA.

Results: The mean platelet count was 6.05 ΁ 1.94 x 1010 in PRPPC, 6.54 ΁ 2.18 x 1010 in BC-PC and 3.84 ΁ 0.77 x 1011 in SDP. In pre lab side filtered PC it was 4.8 ΁ 2.43 and 3.22 ΁ 1.80 in post sample. In bedside filtered PC pre sample had 7.5 ΁ 2.25 and post sample had 5.60 ΁ 2.48 platelet x 1010/unit. RANTES level at one hour was 1210 ΁ 560pg/ml in PRPPC and 1384 ΁ 463 in BC PC. At 112 h it was 1617 ΁ 451 and 1949 ΁ 134pg/ml respectively. In SDP 0 day level was 1850 ΁ 278 pg/ml and was >2000 on 5 th day. In pre-storage lab side filtered PC RANTES was 1035 ΁ 496pg/ml and in post storage sample 310 ΁ 508pg/ml. With bed side filters pre sample showed 1243 ΁ 832 and in post sample 556 ΁ 748pg/ml.

Conclusion: Platelet count is obviously maximum in SDP but more in BC-PC compared to PRP and filtered PC. The concentration of RANTES increased continuously from one hour upto 5 days of storage in all PCs. After 65 hrs BC PC showed higher levels of RANTES compared to PRP-PC. The concentration was least in pre-storage filtered PC. Comparison of different PCs shows that filtered PRP- PCs appear to be the best in terms of low RANTES to prevent allergic reactions.

Should NAT be mandatory?

Shraddha Deogaonkar, MH Chauhan

Dattaji Bhale Blood Bank, Aurangabad, Maharashtra, India

Background: Transfusion safety begins with healthy donors. Incidence of Transfusion Transmitted Infections (TTIs) is 2.4% in India. Early detection of TTIs prevents its secondary transmission and also improves donor health. This can be achieved with the help of Nucleic acid amplification testing technique (NAT), which detects the infectious agent in the window period.

Aim: To study the utility of NAT in preventing TTIs.

Materials and Methods: All ELISA negative units were tested for HBV, HCV, HIV 1 and 2 using NAT. NAT reactive donors were notified telephonically by a counsellor and their follow up serological retesting was done at 1, 2 and 6 months from the point of NAT reactivity.

Results: We evaluated 34,116 donors over a period of 29 months and found 51 ELISA negative donors reactive by NAT. Out of these 51 donors, 50 were reactive for HBV and 1 was reactive for HCV. We approached 51 reactive donors by phone calls, out of which 30 (58.82%) responded. We sampled them for ELISA and found 9 (30%) donors turning serologically reactive over a period of 2-6 months. We could not get the follow up samples of the rest of them (21). Finally 9 out of 34,116 donors (0.026%) could be detected reactive only by the use of NAT. Single blood unit is converted to 3 components and hence we could save 27 people from contracting TTI.

Conclusion: Our study emphasizes the importance of screening every unit of blood by NAT in addition to the routine serological testing.

Therapeutic plasma exchange: Review of the results

Rashmi Sood, Sushma Rani, Vijay Kumar, Deepak Kumar, Tarun Kumar

Department of Transfusion Medicine, Immunohematology and Blood Bank, Saket City Hospital, New Delhi, India

Background: Therapeutic plasma-exchange (TPE) is used as primary as well as adjunctive therapy in treatment of several category of diseases. [1],[2] Technological innovations and the adoption of evidence-based indications in the field of medicine have led to the establishment of guidelines on the clinical indications for the use of therapeutic apheresis. [3]

Aim: Review of the outcome of the consecutive TPE procedures performed on various categories of patients.

Methods: Retrospective analysis and evaluation of the TPE procedures performed at our centre was done for a time duration of July 2013-2015 June.

Results: A total of 71 TPE procedures were performed in patients (2 males and 8females, with mean age of 57.2 ΁ 5 years). Three of the 10 patients had Guillain-Barrι syndrome (GBS), 1 patient had anti glomerular basement membrane CKD disease (Good pasteurs Syndrome), 2 patients had Wegeners Granulomatosis (Systemic Vasculitis cANCA positive), 2 had Myasthenia Gravis, 1 Chronic Inflammatory Demyelinating Polyneuropathy (CIDP) and 1 patient was given TPE treatment for SLE. Out of the total, 9 patients belonged to indication category 1 and one patient belonged to indication category 111.

Fresh frozen plasma (FFP) was used as replacement fluid. Complete response (CR) was achieved on 8 patients (80%) in of the cases. Complications were detected in none of the TPE procedures. Adverse events (AEs), were seen in 5.6% of all procedures. None of the patients died from any complication or adverse event. AE occurred as three as Grade-I (mild -no treatment required and procedure continued) and one patient as Grade-III (severe- treatment required, procedure had to be finished). The most common AE were nausea/vomiting, hypotension, pruritus and abdominal pain.

Conclusions: TPE is effectively and safely carried out for various diseases.


1. MedlinePlus. "Goodpasture Syndrome". U.S. National Library of Medicine. Available form: [Last retrieved on 2013 Apr 07].

2. Yazdi MF, Baghianimoghadam M, Nazmiyeh H, Ahmadabadi AD, Adabi MA. Response to plasmapheresis in myasthenia gravis patients: 22 cases report. Rom J Intern Med 2012;50:245-7.

3. McLeod BC. Therapeutic apheresis: history, clinical application, and lingering uncertainties. Transfusion 2010;50:1413-26.

An audit of red blood cells order and utilization in a tertiary care hospital

K Vijaykumar, Anila Mathan

Department of Transfusion Medicine, Apollo Speciality Hospital, Chennai, Tamil Nadu, India

Background: In a recently started multispecialty hospital the Blood Bank set up an audit to evaluate the efficiency of utilization of packed red cells and to assess the transfusion practice and appropriateness of requests for transfusion.

Aim: To calculate probability of transfusion (%T), Transfusion index (TI) and the crossmatch to transfusion ratio (C/T ratio) for a one year period to evaluate changes that may be needed in blood transfusion ordering practice.

Methods: Data collection included number of patient requests, number of units of packed red blood cells cross-matched and the number of units transfused. Data was retrospectively collected from May to Oct 2014 and prospectively thereafter to April 2015.

Results: Blood was requested for a total of 2824 patients for whom a total of 5719 units were cross matched and 2696 units were issued. The probability of transfusion in the Surgical units (Neurosurgery, Cardiothoracic Surgery, Gastrointestinal Surgery, Orthopaedics, Obstetrics) ranged from 28% in Obstetrical patients to 91% in Cardiothoracic Patients. The number of requests from Medical Specialties (General Medicine, Nephrology, Emergency and ICU) was smaller and transfusion probability was much higher, 79-90%. The transfusion index and the C/T ratio paralleled the probability for transfusion. 47% of the 5719 cross matched units of red cells were utilized with an overall transfusion index of 0.95 and a C/T ratio of 2.12.

Conclusion: It is clear from the high probability of transfusion and high transfusion index that even though this tertiary care hospital and its blood bank are newly started, the critical checks and balances are already in place to ensure that wastage of blood due to over ordering is minimal. We plan to use the audit of blood utilisation to develop strategies for developing Maximum Surgical Blood Order Schedules (MSBOS) procedure wise and to see whether the Group and Save (G&S) can be further used to further bring down the C/T ratio. Continuing monitoring and discussion with Clinicians and the Transfusion Committee will contribute to make the system more efficient.

Determination of select biochemical and hematological reference intervals in Indian voluntary blood donors

Preeti Chavan, Vivek Bhat 1 , Shashank Ojha 2 , Minal Poojary 2 , Sumathi 2 , Mrinal Ganage 2 , Manik Tiwari, Swati Waykar, Umakant Gavhane, Babu Pillai

Departments of 1 Microbiology and 2 Transfusion Medicine, Composite Laboratory, Advanced Centre for Treatment Research and Education in Cancer, Tata Memorial Centre, Navi Mumbai, Maharashtra, India

Background: Major clinical decisions depend on biochemical test reports based on reliable reference intervals. Reference values are used to describe the dispersion of variables in healthy individuals. They are usually reported as population-based reference intervals (RIs) comprising 95% of the healthy population. They may vary significantly in different populations and cultures depending on the dietary preferences, race, geography as well as socio-economic status. Most of the reference intervals provided by various manufacturers for their reagents and methods are based on European or American population (Caucasian) studies and recommend laboratories to establish their own reference intervals. However, very few Indian studies are available on this subject.

Aim: Aim of this study is to establish 95% reference interval for select biochemical parameters in Indian voluntary blood donor population and to correlate the above with respect to physiognomic parameters like gender.

Methods: Blood samples from 350 voluntary blood donors (286 males and 64 Females) were analyzed for Complete Blood Count (CBC) and Serum B12, Serum Folate and Serum Ferritin values on Advia 2120i fully automated 5-part differential hematology analyzer and Architect plus i 1000 SR fully automated immunoassay analyzer. The biological reference interval (BRI) was calculated between the 0.025 and 0.975 percentile and represents the central 95% confidence limit.

Results: Reference ranges obtained for White Blood Cells (M: 4.5-11.0 & F: 5.6-11.3 X 109/L), Red Blood Cells (M: 4.25-6.6 & F: 3.8-5.3 X 1012/L), Hemoglobin (M: 13-18.4 & F: 11.9-15.2 g/dL), Platelet Count (M: 137-379 & F: 185-439 X 109/L), MCV (M:77.7-101.6 & F: 78.2-98.3 fL), Serum Vitamin B12 (M: 100-500 & F: 113-481 pg/mL), Serum Folate (M: 2.8-18.8 & F: 3.3-16.8 pg/mL), Serum Ferritin (M:11.2-202.8 & F: 11.1-84.4 ng/mL).

Conclusion: Compared to the existing reference interval platelet count in Male donors was found to be lower. Similarly limit of MCV in both male and female donors was lower than existing intervals. Serum levels of Vitamin B12 and Ferritin showed much lower ranges in Indian voluntary blood donors than the current ranges used. This difference may be attributed to genetic differences as well as dietary preferences in our population. However, further studies with larger sample size need to be conducted to substantiate the above findings.

Transfusion related adverse reactions reported in tertiary care centre: A step towards hemovigilance

Rajni Bassi, Kanchan Bhardwaj, Shikha Aggarwal, Kusum K Thakur

Department of Transfusion Medicine, Government Medical Collge, Patiala, Punjab, India

Introduction: Transfusion of blood and blood products is a double edged sword, so should be used judiciously. Though blood transfusion can be life-saving, it can also lead to certain adverse reactions which can be relatively mild to severe life threatening. Knowledge about various types of blood transfusion reactions will help not only in their early identification and management, but also in taking adequate measures to prevent the same.

Aim: To determine the frequency and types of adverse transfusion reactions in recipients of blood and blood components.

Materials and Methods: Prospective study from January 2014 till April 2015 was done. Transfusion reactions reported to the Department of Transfusion Medicine, Government Medical College, Patiala, were recorded and analyzed on the basis of their clinical features and lab tests.

Results: During the study period 25099 units of blood and blood components were transfused and 100 transfusion reactions were reported. The overall incidence of ATRs in our study was 0.4%. The most frequent were febrile non- hemolytic transfusion reactions (FNHTR) (73%) followed by allergic reactions (24%), bacterial infections (1%), hypotension due to ACE inhibitors (1%) and hemolytic (1%). Incidence of transfusion reactions due to platelet concentrates transfusion was 0.5%, packed cells whole blood transfusion 0.43% followed by FFP 0.04%.

Conclusions: The majority of the types of reactions observed were FNHTRs followed by allergic reactions. Adequate skilled and dedicated manpower, reporting of all adverse events, fully functioning hospital transfusion committee with continuous medical education to medical and paramedical staff will definitely help in strengthening hemovigilance system and reducing the incidence of adverse TRs to minimum.

A case of suspected low avidity B antibody

J Sanal, N Sharma, P Jain, R Thakkar, P Desai, SB Rajadhyaksha

Department of Transfusion Medicine, Tata Memorial Centre,Navi Mumbai, India

Introduction: Weak antigen expression and decreased antibody synthesis is a known phenomenon in oncology patients with hematolymphoid malignancy and solid tumours. A case report from a tertiary care oncology centre is being presented here.

Case Details: 54 year old female with a diagnosis of breast cancer came to our center in April 2015 for further management. Her blood sample was received for blood grouping.

Initial grouping was performed using automated column agglutination technique (CAT) system & showed cell group as 'O Rh positive' and serum group as 'B' group. Patient's antibody screen using commercial screening cells was negative. Direct Antiglobulin Test and auto control was negative. A fresh sample was requested and on repeat grouping with automated CAT system and conventional tube technique, the discrepancy persisted.

An adsorption - elution study using patient's RBCs and anti B serum were carried out, which was negative for the presence of 'B' antigen.

Detailed clinical history was elicited and it was found that patient did not have any past history of transfusion, though the blood group was known to her as 'O positive'.

Co morbidities: Hypothyroidism diagnosed 6 months back and anemia 1 month prior to admission.

Medication History: Thyronorm 75 ΅g for 6 months, Anastrazole 1 mg/day for 1 month, oral iron and Vit B12 (I.V) for 1 month.

Further serological workup was done to confirm the blood group:

Altering cell to serum ratio (1:5) using 'pooled in-house B cells' with extended incubation at room temperature again showed negative result ('B blood group' on reverse grouping)

Patient's serum was incubated with individual B cells for 2 hours at room temperature and 40C along with auto-control. No agglutination was observed at room temperature but at 40C, agglutination ranging from weak to 3+ was seen. Auto control was negative

Salivary testing for 'secretor status' showed presence of 'H antigen' and absence of 'A' & 'B' antigen.

Considering all the lab results together this patient was reported as 'O Rh positive'.

Discussion: ABO blood group discrepancies can be multifactorial and are categorized as below:

Weak or missing antibody

Weak or missing antigen

Extra red cell reactivity

Extra serum reactivity.

As per one Indian study on 1,04,010 donors, blood group discrepancies was found in 51 cases (0.04%), of which 30 (58.8%) cases were low avidity anti-B antibodies. In this present case also the weakening of anti-B activity can be due to low avidity antibody due to underlying disease.

Conclusion: Oncology patients are a special subset of patients who may present with discrepancy in cell and serum grouping hence a complete immuno-hematological work up is warranted. Prompt identification of the discrepancy is pivotal to clinicians to meet the transfusion demands.

Ethical dilemma in disposal of unused cryopreserved hematopoetic stem cells in a bone marrow transplantation centre

Minal Poojary, S Devadas 1 , S Parab, S Ojha, S Kannan 2 , S Rajadhyaksha, N Khattry 1

Department of Transfusion Medicine, Tata Memorial Centre, 1 Department of Haemato-Oncology, Bone Marrow Transplant Unit, Tata Memorial Centre, 2 Department of Biostatistics, Advanced Centre for Treatment Research and Education in Cancer, Tata Memorial Centre, Navi Mumbai, Maharashtra, India

Background: Hematopoietic Stem cell units (HSCU) are cryopreserved prior to both Autologous Hematopoietic Stem Cell Transplant (HSCT) and as Donor Lymphocyte Infusion (DLI) in Allogeneic HSCT for Donor. Many times HSCU's remain either unused or partially used imposing spatial, economic and logistic issues. Further, leading to increased ethical burden on HSCU depositories. Due to lack of adequate data regarding duration of useful storage and undefined shelf life of HSC there is lack of general consensus for disposal of HSCU's.

Aim: To debate the duration of usefulness of storage of Hematopoietic Stem Cells (HSC) and define consensus for their appropriate disposal.

Methods: We conducted a retrospective analysis of unused Hematopoietic Stem Cell Units (HSCU's) in 435 patients planned for either Autologous (n = 209) or Allogeneic Stem Cell Transplantation (n = 196) at our Bone Marrow Transplantation Centre between November 2007 to January 2015.

Results: Our cohort consist total of 2457 HSCUs, out of these 1728 HSCU's are stored for Autologous HSCT and 729 HSCU's stored for Donor Lymphocyte Infusions (DLI) after Allogeneic HSCT. For Autologous HSCT median CD34 cell dose infused was 5.28 x 106 cells/kg and for patients requiring DLI median CD3 cell dose infused was 2.05 x 107/kg. In 12.5% of patients planned for Autologous HSCT never underwent reinfusion and 80% of patients who underwent Allogeneic HSCT never received DLI. 41% of HSCU for Autologous HSCT remained with us with second Autologous HSCT being done in only 4 patients. 91% of HSCU stored for DLI remained unused, with only minimal usage after one year from Allogeneic HSCT. All the HSCU's are still stored with us.

Conclusion: Duration of storage of unused HSCU needs to be defined for proper disposal of the same. Till then storage of HSCU will remain a major ethical and logistic challenge.

Analysis of rejection criteria of platelet pheresis donors

Jignesh J Desai, Abhay Jhaveri, Rinku Shukla

Surat Raktadan Kendra and Research Centre (NABH Accredited Regional Blood Transfusion Centre), Surat, Gujarat, India

Background: Blood bank has a policy to collect blood from voluntary, non-remunerated, low risk, safe and healthy donors. Donors are selected according to the donor selection criteria mandatory as per the Drug and Cosmetics Act, 1945 amended time and again, published by the Ministry of Health in the Gazette of India.

Aim: To find out the reasons for deferring the donors who were enrolled for platelet pheresis during last five years at our blood centre and analyze them in detail.

Methods: All the details of donors who come to our blood centre are entered in our blood bank management software, whether they are accepted as donors or deferred. We found out the donors who were enrolled for platelet pheresis procedure during last five years (January 2010 to December 2014) from the software, transferred the data to Microsoft Excel Worksheet and analyzed the data.

Results: Total 2514 donors were screened for platelet pheresis procedure. Out of them, 1976 donors (78.60%) were selected and 538 donors (21.40%) were deferred due to various reasons. Out of these 243 donors (45.15%) were rejected as their veins were not proper for pheresis procedure. 79 donors (14.68%) were rejected as their platelet count was <200000/cmm (though the most guidelines allows us to accept the donor if platelet count is ≥150000/cmm, for donor safety we have kept the limit to 200000/cmm). 66 donors (12.27%) had hypertension, 32 donors (5.95%) had hemoglobin <12.5 g/dL. We try to analyze the cause of anemia in such donors and provide oral iron and folic acid tablets to iron deficient donors. By this, we bring them back in donor pool. 27 donors (5.02%) had taken either antibiotic or aspirin or other anti-platelet agent within last 72 hours. 19 donors (3.53%) were first or second degree blood relative. For last two years, we have the facility of irradiator. So in case of platelet refractoriness, in HLA matched blood relative donors, we accept first degree relatives and irradiate the product prior to issue.

Conclusion: Our data suggest that the most common reason for deferral for platelet pheresis is improper vein, followed by low platelet count. Blood banks with irradiation facility can accept the first or second degree blood relatives and increase donor pool.

Anti-G, a masquerading antibody: Case report

S Gupta, A Maheshwari, P Rajani, M Bajpai

Department of Transfusion Medicine, Institute of Liver and Biliary Sciences, New Delhi, India

Introduction: G antigen is present on most red cells expressing Rh D or C antigens. It isalmost always absent from cells that are D and C negative. Rh D negative patients can produce anti-G antibody following transfusion of Rh D negative C positive red cells. Anti-G is a clinicallysignificant antibody which may cause hemolytic disease of fetus and newborn and hemolytic transfusion reactions, but the occurrence is rare and outcome seldom severe. It may bemistaken for anti-D+C. It is important that D negative pregnant woman with anti-G or anti-C+G, receive anti-D immunoglobulin to prevent them making anti-D.

Case Report: A 47 year old female presented with HCV related Chronic liver disease. She required packed RBC transfusion for correction of anemia. She had two living children and a history of one abortion. She had received multiple transfusions in the past. Her blood group was O Rh D negative. Her husb d's blood group was B Rh D pos. Her antibody screen was positive. On testing the sample against antibody identification panel, anti D+C was suspected. As anti -G mimics anti -D+C, differential adsorption elution method was performed to identify anti-C, anti-D and anti-G. The pati t's ser was first adsorbed with D+C- cells to adsorb anti- D and G (if present) on the cells, and leave out anti-C in the adsorbed serum. The adsorbed serum gave a 3+ reaction with D-C+, D+C+ cells and no reaction against D+C-, D-C- cells, confirming the presence of anti-C. Elution was done on these D+C- cells using the acid elution technique. Then this eluate which contains no anti-C was then adsorbed with D-C+ cells, to isolate anti-G if present. These adsorbed D-C+ cells were eluted in the same manner and the eluate gave a 4+ reaction against D+C-, D-C+, D+C+ reagent cells and no reaction against D-C cells, confirming the presence of anti-G. The adsorbed eluate gave no reaction against D+C-, DC+, D+C+, D-C- reagent cells, ruling out anti-D. Thus sequential adsorption and elution with D+C and D-C+ cells confirmed that patient had anti C and anti-G antibody, and ruled out anti-D.

Conclusion: Double elution method is useful in differentiating anti-C, anti-D and anti-G. The procedure is quite time consuming one, requiring a panel of select cells and undivided attention of the performer, but then it is important to differentiate anti-G because otherwise D negative pregnant women with anti-G or anti-C+G may be deprived of anti-D immunoglobulin prophylaxis, if antibody specificity is not identified.

Anti-M: Report of two cardiac cases with review of literature

Sadhana Mangwana, K Pushpa, S Sharma, D Arya

Department of Blood Transfusion Services, Sri Balaji Action Medical Institute, New Delhi, India

Anti-M is a naturally occurring antibody reacting optimally at 4-250C. Anti-M consists of Immunoglobulin M (IgM) antibody but considerable number may have IgG component. Anti-M antibody is more common in children than in adults. Most Anti-M are not active at 37΀C and generally be ignored in transfusion practice. However, this antibody, if present in an individual, can lead to a problem in immunohematology laboratory.

We report 2 cases of clinically significant anti-M antibodies that came to hospital with history of chest pain and shortness of breath and diagnosed as coronary artery disease. One case presented as cross match incompatibility and other showed discrepancy between forward and reverse ABO grouping thus creating diagnostic difficulty for blood bank staff. We reviewed the literature to find out the significance of cross-match incompatibility and blood group discrepancy.

Seroprevalance and possible correlation of transfusion transmitted diseases with Rh type and abo blood group system

Deepti Agarwal, AK Gupta, Neetu Singh, Vibha Singh, Ajay Sagar, AK Verma

Department of Transfusion Medicine, Employees State Insurance Postgraduate Institute of Medical Sciences and Research, New Delhi, India

Background: Screening of blood is mandatory for transfusion transmitted diseases and is routinely done in the blood banks. Since, literature has now depicted that some blood groups are more prone for seropositivity of one or other infections; we tried to analyze the donor populations at our institute to have group wise distribution and infectious markers for any association if any.

Aims: Our aim was to find prevalence and distribution of various blood groups in our donors and to identify seropositivity of various infectious markers amongst them and to find any association.

Materials and Methods: This retrospective study was conducted at the Department of Transfusion medicine, E.S.I Hospital, Basaidarapur, New Delhi for a period of one year from January 2014 to December 2014.

All voluntary and replacement donors reporting to the blood bank were incorporated and analyzed. Blood group of all donors was recorded and analyzed accordingly. Screening for HIV-1 & 2, HBsAg, and HCV were done by using 4 th .

Generation appropriate Enzyme-linked immunosorbent assay (ELISA) technique using micro-ELISA kit. The venereal disease research laboratory (VDRL) test was used for estimation of syphilis infection. The rapid malaria diagnostic kit was used for determining the malarial infection.

The data entry was carried out using Microsoft office excel worksheet and was analyzed by percentage and comparison.

Results: A total of 7324 (n) donations were done during the calendar year of 2014. The blood group prevalent amongst the donor population from most common to less common was B>O>A>AB (35.07>26.34>22.41>10.28% respectively).

Seroprevalence of various infectious markers was found to be HIV (0.2457 %), HCV (0.477%), HBsAg (1.2834 %) and syphilis (0.0136 %). HIV seropositivity was found to be maximum in A>O>AB>B (0.36% >0.30% >0.26% >0.15%). HBsAg seropositivity was maximum seen in Blood group A (positive) followed by AB (Positive), O (Negative) and A (Negative) group in order of decreasing frequency. HCV showed maximum seropositivity in negative blood groups (0.46% in positive vs. 1.10 % negative groups). In our study, only 1 case of syphilis was detected and none of our donors were found reactive for malaria.

Conclusion: Is the most prevalent blood group least predisposed to TTI? Our study results point in this direction! Though our data size for donors was quite large, the number of seropositive cases were limited likely due to our rigorous screening protocols and procedures. Hence, multicentre studies should be encouraged in this regard to know the exact association for infectious markers in relation to various blood groups; as it happened with Gastric carcinoma and gastritis for blood group A and O respectively.

Successful management of refractory dialysis independent Wegener's granulomatosis with combination of therapeutic plasma exchange and rituximab

Sheetal Malhotra, Hari Krishan Dhawan 1 , Ratti Ram Sharma 1 , Neelam Marwaha 1 , Aman Sharma 2

Department of Transfusion Medicine, Maharishi Markandeshwar University, Mullana, Ambala, Haryana, Departments of 1 Transfusion Medicine and 2 Internal Medicine, PGIMER, Chandigarh, India

Background: Wegeners Granulomatosis (WG) is an autoimmune, anti-neutrophil cytoplasmic antibody (ANCA) mediated necrotizing vasculitis with a mortality rate of 90% if left untreated. Treatment relies on a combination of immunosuppressive drugs and tapering regimen of glucocorticoids. Therapeutic plasma exchange (TPE) has been effectively used in the management of systemic vasculitides and autoimmune disorders based on its principle of removal of pathogenic autoantibodies and immune complexes. Few case reports and randomized controlled trials have demonstrated the use of TPE as a second line treatment for WG. As per ASFA guidelines, TPE is category I indication for ANCA associated WG in dialysis dependent cases and category III indication for dialysis independent cases.

Aim: We report the successful use of TPE in combination with rituximab in achieving remission in a patient with WG (dialysis independent) not responding to conventional therapy.

Report of a Case (Methods/Results): A 58-year old male, known case of ANCA positive WG for the past two years presented with disease exacerbation for the last three days. For the treatment of WG, he had received 18 pulses of high dose cyclophosphamide at every 4 weeks interval. During this time, he experienced three relapses in disease activity after initial improvement in clinical symptoms. Patient was dialysis independent. In addition, he was a diagnosed case of pulmonary and abdominal tuberculosis, and was on anti-tubercular therapy for two months. Investigations revealed anaemia (6.4g/dl), leukocytosis (10600/ul), azotemia (serum urea- 84.4 mg% which shot upto 218.1mg%, serum creatinine - 7.5mg%) and mild tranaminitis (SGOT-36U/L, SGPT-52.3 U/L). For managing the current disease exacerbation, therapeutic plasma exchanges were planned along with intravenous rituximab and oral corticosteroids (prednisolone 80 mg/day). A total of twelve TPE procedures were performed at one or two day's interval over a month. Due to low hemoglobin, TPE kit was primed with a packed red blood cell (PRBC) unit for the initial five procedures. One plasma volume exchange was done in all the procedures with replacement fluids as 5% human serum albumin and normal saline. Blood group specific fresh frozen plasma was also used in three of the procedures. The patient had no complications during the procedures except in one in which he had mild shivering at the end of the procedure. It subsided on giving intravenous chlorpheniramine and hydrocortisone. With treatment, clinical, haematological and biochemical parameters improved substantially. Fever, conjuctival bleed and epistaxis subsided and remission was achieved.

Conclusions: TPE along with rituximab and steroids combination therapy is a useful therapeutic option for patients in whom the disease is refractory to cyclophosphamide. However, TPE combined with immunosuppressive drugs should be used under close monitoring with proper dose schedule adjustments alternating with TPE procedures.

An audit of cryoprecipitate usage in a tertiary care hospital

C Jayasree, B Abhishekh, RG Kulkarni

Department of Transfusion Medicine, JIPMER, Puducherry, India

Background: Cryoprecipitate is a rich and concentrated source of Factor VIII, VWF, Fibrinogen, Factor XIII and Fibronectin. It contains other clotting factors as well, but at significantly low levels (20-30%) than that present in fresh frozen plasma. The recommended indications of usage of cryoprecipitate are very limited and mainly includes Haemophilia a, Von Willebrand's disease, Afibrinogenemia/Dysfibrinogenemia, FACTOR XIII deficiency etc. It is also being widely used for other indications such as for controlling bleeding following surgery or trauma with no demonstrated specific derangement of VWF, Fibrinogen, Factor VIII or Factor XIII. Even in circumstances under which it is used there are varied irregularities in dosage.

Aim: To audit the usage of cryoprecipitate in our tertiary care teaching hospital over a period of six months retrospectively so as to formulate guidelines for its usage in the hospital.

Methodology: A retrospective collection of all requests made for cryoprecipitate from December 2014 to May 2015 was done by noting the clinical diagnosis, the indication for transfusion and the number of units issued. Using the Hospital Information System pre and post transfusion investigations including Prothrombin time/INR, APTT, platelet count, fibrinogen level etc. of each patient were evaluated.

Results: 377 units of Cryo were issued of which 201 (53.32%) were for bleeding following trauma/ surgery/delivery/abortion; 93 (24.67%) were for DIC, 48 (12.73%) for Haemophilia A; 12 (3.18%) for Haemophilia B;9 (2.39%)for VWD;6 (1.59%) for TTP;6 (1.59%)for Factor XIII deficiency;1 (0.27%) for Afibrinogenemia and 1 (0.27%)for organic aciduria with deep vein thrombosis.

Conclusion: The misuse of cryoprecipitate has been well documented in literature across the globe. Auditing the usage at the hospital level helps in identifying instances wherein the use of the component is not optimised and hence intervening purposefully. This study throws light on the dearth of commendable guidelines and consensus among treating physicians across specialities about cryoprecipitate usage and that it's the need of the hour for continued education.

Modes of leucoreduced red cell preparation at RBTC-GTB hospital

Surender Kumar, Anju Gupta, Divya Bansal, Bharat Singh

Department of Blood Bank, UCMS and GTB Hospital, New Delhi, India

Background: Blood bank is an integral part of any modern health care institution. In the current era of evidence based medicine, judicious use of safe blood or its component in appropriate clinical setting is essential. Leucoreduction is a process by which leucocytes are removed from donated blood. Leucoreduced products can be obtained by centrifugation and filtration, buffycoat removal, freezing and deglycerolization, washing of RBCs with saline, etc.

Aims and Objective: To study the effectiveness of various modes of leucoreduction.

Materials and Methods: The blood collected at our centre from January 2014- December 2014 was subjected to leucoreduction by various methods like buffycoat removal method for top and bottom (T&B) bags without filters and centrifugation and filteration method for T&B bags with integral filters by using refrigerated centrifuge machine and optipress system. Post processing leucocyte count was done by Coulter hematology analyzers.

Results: Total blood collected during the period was 32401, out of which 12908 (39.84%) were processed for component preparation. 9722 bags (75%) were subjected to leuco-reduction by buffy coat removal method and the remaining 3186 bags (25%) were top and bottom (T&B) bags with integral filters, leuco-reduced by centrifugation and filtration method. The reduction in leucocytes count achieved was approximately 50% by buffy coat removal method, whereas with centrifugation and filteration with bags having integral filters was 100% (i.e. leukocytes count <5X10^6/unit).

Conclusion: Use of filtration method for bags with integral filters is an optimal method of leuco-reduction. However, use of integral filters is costly, but must be used at least in patients who require multiple blood transfusions.

Prevalence and changing trends of transfusion transmitted infections amongst blood donors in East Delhi

Divya Bansal, Akansha Rawat, Preeti Diwaker, Priyanka Gogoi, Bharat Singh

Department of Transfusion Medicine, UCMS and GTB Hospital, New Delhi, India

Background: Transfusion transmitted infections (TTIs) are major problem associated with blood transfusion. Most common TTIs are HIV, Hepatitis B, Hepatitis C, Syphilis, and Malaria. Effective donor screening protocol and accurate estimates of risk of TTIs are essential for monitoring the safety of blood supply.

Aims and Objective:

To compare the prevalence of TTIs in voluntary and replacement donors

To study the changing trends of various TTIs amongst blood donors.

Materials and Methods: The study was an audit of the records of all donations done in GTB Hospital, RBTC (East Delhi) from January 2008 to December 2013. Completely filled donor forms along with TTIs records were analyzed for each case.

Results: Out of total 1,88,171 donations, 1,38,229 (73.45%) were voluntary and 49,942 (26.55%) were replacement donations. The overall prevalence of HIV, HBV, HCV, Syphilis and Malaria were 0.34%, 1.62%, 0.71%, 1.76%, 0.05%, respectively. On studying the trends, there was decrease in number of seropositive cases for HIV and syphilis in both voluntary donors (VD) and replacement donors (RD). Seropositivity for Hepatitis B has considerably decreased in VD, but there is minimal change among RD. The seropositivity for Hepatitis C has increased in RD and reduced among VD.

Conclusion: TTIs are more frequent in RD than VD. The increase in public awareness regarding voluntary blood donation, meticulous donor screening and counseling can help in reducing the prevalence of TTI.

Phenotypic profile of ABO, Rhesus and Kell blood group systems in 14261 healthy blood donors at regional blood transfusion center: Guru Teg Bahadur Hospital, North India

Ruchi Rathore, Vandana Puri, Bharat Singh

Introduction: ABO and Rhesus (Rh) blood group systems are the 2 major blood group (BG) systems in red cell serology. But unlike the ABO system the Rh blood group system is more complex consisting of 49 antigens expressed on chromosome 1. Despite proper blood grouping and cross matching the recipients infrequently get alloimmunized resulting in antibody production against the Rh or other minor blood group antigen. Since complete phenotyping is not possible due to financial constraints in a developing country like ours the Rh phenotyping plays a crucial role in preventing alloimmunization of recipients.

Aim: We conducted this study to observe the frequency of Rh, ABO and Kell blood group antigens in healthy blood donors and thereby generate data for making blood donor panels, as an effort to further reduce the chances of alloimmunization in our population.

Materials and Methods: ABO, Rh and Kell phenotyping is performed using Gel card method (diamed) in our blood bank, regional blood transfusion centre, GTB Hospital, New Delhi. The BG antigen determination using this method has been performed in 14261 healthy continuous voluntary and replacement donations since January 2012 to December 2014 so far. For the purpose of this study the BG antigen data on ABO, Rh (D) and Kell for the last 3 years was collected from the records, analysed using SPSS version 22 software and the results were compiled accordingly.

Results: Among the ABO BG system B+ was the most common phenotype and AB- was the least common ABO phenotype. Of the total 14261 donors, 14067 (98.6%) Were Rh D+ donors and 194 (1.4%) were RhD- donors. Of all the Rh antigens 'e' was the most common antigen accounting upto 96.6% followed by C (90.3%), and E (19.8%) was the least common antigen. Only 1.9% of all the donors were kell antigen positive.

Phenotype frequency of Rh and Kell blood group system among voluntary blood donors in rotary blood bank, New Delhi, India

Asha Kaul Bazaz, Kinjal Upadhyay, Anju Gupta

Background: Transfusion medicine has been constantly evolving through the years with improved technologies that enhance the capability of giving Safe blood to patients. Although blood transfusions can Save lives, but they are not without risk. The most common and most serious is the hemolytic transfusion reaction caused by antibody incompatibility. Knowledge about the frequency of red cell antigen phenotypes is very important to minimize risks of alloimmunization in multi transfused patients. After ABO & Rh blood groups the K antigen is very immunogenic in stimulating antibody production.

Aim: In this study we determine the phenotype frequency of Rh and Kell blood group system among Voluntary blood donors. It also allows us to improve transfusion services providing safe blood to needy patients.

Methods: We analyzed the results of 12413 Voluntary blood donors for Rh phenotype and K antigen tests over a period of 6 months from November 2014 to May 2015. Red cell antigen typing for Rh and Kell was performed on a fully automated immunoheamatology analyzer NEO GALILEO: Immucor Inc. using IgM monoclonal antisera by Micro plate Hemagglutination method and few samples for Rh typing were performed by (CAT) Column agglutination technology on Autovue a fully automated system Ortho Clinical Diagnostics, Johnson and Johnson.

Results: Out of 12413 blood donors 94.1% were found RhD positive while 5.87% were found to be Rh negative. The frequency of C antigen was 83.6%, c antigen was found to be positive in 56.6%, E antigen was 21.6%, e was 95.4%.In the KELL blood group system 2% were found as K antigen positive while as 98% were k antigen positive.

Conclusion: The phenotype frequency of Rh and Kell system in our donors helps us in giving antigen negative blood to the patients where clinically significant antibodies are identified in patient's serum.

Microbial contamination of blood and blood products in a tertiary care oncology set up

Vivek Bhat, Shashank Ojha Preeti Chavan, Rohini Kelkar, Sanjay Biswas, Naina Baraskar

Advanced Centre for Training, Research and Education in Cancer, Tata Memorial Centre, Navi Mumbai, Maharashtra, India

Introduction: Microbial contamination of blood and blood products is a major risk factor associated with morbidity and mortality in transfused recipients. Microbiological safety monitoring is an important component of ensuring the safety and quality of blood products.

Aims: This study aims to determine the spectrum of microorganisms recovered from blood and blood products over a 3 year study period in our tertiary care oncology centre.

Methods: The following donor blood products were collected and sent to the microbiology laboratory for microbial cultures. Peripheral blood stem cells (PBSC) meant to be transfused into hematopoietic stem cell transplant (HSCT) recipients was collected in the COBE Spectra or Fresenius apheresis systems. Segments from the PBSC products (pre -cryopreservation), after adding DMSO (post-cryopreservation) and before infusion (post -thaw) were cultured. In addition cultures from other blood products like random donor platelets (RDP), single donor platelets (SDP), fresh frozen plasma (FFP), packed red blood cells (PRBC), whole blood, granulocyte concentrates and bone marrow harvests were also included in the study. All samples were cultured in the microbiology laboratory as per standard microbiological procedures and susceptibility testing performed if clinically indicated.

Results: A total of 1652 samples of various blood products were cultured, of which 46(2.8%) were culture positive. The number of culture positives vis-a vis the number of products cultured were as follows: PBSC (pre & post cryo and post thaw) 36/608; SDP (4/309); FFP (1/135); RDP (0/170); PRBC (3/290); whole blood (1/131); Granulocyte concentrates(2/6) and Bone marrow harvests (1/3). The organisms that were more frequently isolated included Klebsiella pneumonaie (12), Coagulase negative Staphylococcus (CoNS-9),  Escherichia More Details coli (6), Proteus spp. (4) &  Pseudomonas aeruginosa Scientific Name Search ). Klebsiella pneumoniae Scientific Name Search  was the commonest organism isolated from PBSCs.

Conclusion: The overall incidence of microbial contamination was found to be 2.8% with Klebsiella pneumoniae and Coagulase negative Staphylococci being the most common.

Evaluation of adverse reactions in blood and apheresis donors

V Patil, S Ojha, K Chawan, A Tirlotkar, M Poojary, S Vimal, S Rajadhyakshya 1

Department of Transfusion Medicine, Advanced Centre for Training, Research and Education in Cancer, Tata Memorial Centre, 1 Department of Transfusion Medicine, Tata Memorial Centre, Navi Mumbai, Maharashtra, India

Background: The increasing demand for blood and platelets constantly challenges blood centers to maintain a safe blood and apheresis donations, but occasionally adverse reactions of variable severity may occur during or after the collection. Adverse reactions have negative impact on donor recruitment and retention. .To assess the rate of adverse reactions in our centre, we analyzed all adverse events among blood and apheresis platelet donors.

Aim: To evaluate frequency and type of adverse reactions in blood and platelet donors.

Methods: Retrospective analysis of adverse reactions was done in blood and apheresis platelet donations during the period of January 2013 to December 2014. These included 4019 blood and 1520 apheresis platelet donors respectively, who were separately analyzed and subsequently compared for common adverse reactions like hematoma, vasovagal reaction (VVR), citrate reaction etc.

Results: Total of 112 (2.78%) adverse reactions were observed in blood donors whereas 102 (6.7%) in apheresis platelet donors. VVR with mild intensity were most commonly observed in blood donors (2.53%) specially in first time donors (1.26%) followed by hematoma (0.25%) whereas hematoma (5.53%) were observed more common in apheresis platelet donors followed by VVR (0.72%) and citrate reactions (0.46%). In apheresis platelet donors only 1 male (0.06%) had grade II VVR. The incidence of VVR and haematoma compared with male versus female in both type of donors was not statistically significant (P > 0.05) but VVR in blood donors compared to apheresis platelet donors was found to be statistically significant. (P value = 0.0001).

Conclusions: Obtaining such data on incidence of adverse events enables to improve safety and comfort of the donor. Also it helps in minimizing negative impact on donor recruitment and retention thereby improving blood donor return rate.

Automated anti-A/anti-B titration in ABO incompatible renal transplant patients: A new breakthrough in Indian blood banking

Sangeeta Pathak, S Singh, T Chakroborty, S Kaushik, R Dubey, V Gangwar 1 , A Gupta 1

Department of Transfusion Medicine, Max Healthcare Institute, 1 Department of Application, Immucor India Private Limited, New Delhi, India

Background: Antibody titer is one of the few tests in blood banking that has proved to be difficult due to the variability of manual dilutions and the subjectivity determining the end-point titer. The AABB Technical Manual states that titration is a semi-quantitative method and is quite technique dependent, because many variables can affect testing results; the procedure is relatively imprecise. The use of automated methods offers the prospect of standardization and reproducibility of results.

Aim: To establish the routine Anti-A & Anti-B titration using a fully automated system and to minimize the risk of variability due to manual dilutions and subjective end-point reading with documentation.

Materials and Methods: In 6 months total 644 numbers of blood samples from renal transplant patients at a tertiary care hospital in Delhi, were tested as, 298 (46%) for anti-A & 346 (54%) for anti-B, specifically IgG antibodies. Testing was carried out by two different methods, 1) Fully automated immunohematology analyzer Galileo-NEO (Immucor-Norcross, GA) and, 2) Semi-automated Bio-Rad ID Gel Card. Samples were collected in EDTA and Plain vials. Results were generated via Solid Phase Red Cell Adherence Technology in Galileo-NEO and by AHG method in Gel card. The instrument fully automate the assay process, including sample preparation, serial dilution, and interpretation, while in Gel Card, manual method of serial dilution & end point reading method was adopted.

Results: In this study of two different automated & semi-automated methods; 483 (75%) results were founded as same titer report. 15% of titer reading were recorded 1 fold dilution increased & 7% were recorded 1 fold dilution decreased in Gel card rather than in Galileo-NEO. 2% of samples also founded as 2 fold dilution increased while 1% samples founded 2 fold dilution decreased in Gel. These total 161 (25%) samples which gave variable reaction were repeated in both method and found 78 (12%) samples giving same result in both with reproducibility in automation while 83 (13%) giving 1 fold dilutions increased.

Discussion: Antibody titrations are important in antenatal evaluations, transfusing ABO-incompatible plasma products, and performing ABO-incompatible organ transplants. As per AABB, The titer is determined from the highest dilution of serum that gives a reaction of 1+, macroscopic agglutination. Variations in technique can cause duplicate tests to give variable results. Serum containing antibody at a true titer of 32 may show variability on replicate tests. [1] It is often seen in study that 1 fold increase & decreased is subject to making serial dilution & reading of reaction individually. The accepted titration methodology is the tube method, Titration using gel column technology may result in titers several dilutions higher than the tube method. Steiner et al. reported antibody titers and scores in gel to be consistently higher than titers and scores in tubes. [2] When a large number of ABO incompatible solid organ transplant programs in the country are seeking Transfusion Medicine for a true titer value, it is better to introduce an automated method of titration in spite of manual method to escape such type of variability in titer assay.


1. Roback JD, Grossman BJ, Harris T, Hilllyer CD. AABB Technical Manual. 17 th ed. Bethesda MD: AABB Press; 2011.

2. Steiner EA, Judd WJ, Combs M, Watkins K, Weissman P, Mann N, et al. Prenatal antibody titers by the gel test. Scientific Abstract, Ortho Clinical Diagnostics, 2002 AABB Annual Meeting Presentation; 2002.

Knowledge, attitudes and beliefs towards voluntary blood donation amongst undergraduate medical students at a tertiary healthcare institution in Uttarakhand, India

Doda Ananya, Negi Gita, Harsh Meena

Department of Pathology, Himalayan Institute of Medical Sciences, Swami Rama Himalayan University, Dehradun, Uttarakhand, India

Background: Blood donors form the backbone of blood transfusion services. Statistics show that there are 234 million major operations in India, 63 million trauma-induced surgeries, 31 million cancer-related procedures and 10 million pregnancy related complications which require blood transfusions. Apart from these there are also disorders like sickle cell anemia, thalassemia and haemophilia that require repeated blood transfusions. Blood donation is the only way of acquiring blood to meet the requirements. Globally, 80 million units of blood are donated each year. India, despite being a country with a population of 1.25 billion, faces a blood shortage of 3 million units every year.

Aims: The objective of this study was to determine the knowledge and attitude of medical students of a tertiary healthcare institution in Uttarakhand, India towards voluntary blood donation.

Methods: This descriptive cross-sectional study was conducted amongst MBBS students at Himalayan Institute of Medical Sciences, Dehradun, Uttarakhand (India). We assessed awareness, knowledge, attitude, perceptions and factors affecting voluntary blood donation in the students by a validated and pre-tested questionnaire. The consented information was analyzed using Microsoft Excel.

Results: More than half of the respondents had never donated blood. Of those that had donated, most were voluntary donors. Approximately two-thirds of total respondents had good knowledge of blood donation. Most students showed a positive attitude towards blood donation. Among those that had donated, males were more than females. Many donated for moral satisfaction in the regularly held blood donation camps at our institution. The reasons why many did not donate were lack of awareness, lack of opportunity due to tight lecture schedule, lack of knowledge and inadequate information about blood collection services along with fear of weakness and fear related to venepuncture. Some students agreed that IEC material and gift items such as T-shirts, stationery, mugs and wristbands would motivate them to donate blood.

Conclusion: The role of youth in voluntary blood donation is crucial to meet the demand of safe blood. It was possible to understand the various factors that influence blood donation which is the basis for donor mobilization and retention strategies. A positive attitude and practice can be improved by inculcating knowledge about voluntary donation amongst medical students so as to recruit regular donors. Owing to ease of access to hospital services, the students organizations in the university should include a blood donation drive in their activities. The university authorities along with the Department of Pathology & Transfusion Medicine should collaborate in promoting voluntary blood donation among the students more actively.

Intra operative Rh D switching in a patient requiring massive transfusion: Case report

K Sujitha, B Abhishekh, RG Kulkarni

Department of Transfusion Medicine, Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India

Introduction: Massive transfusion in acute bleeding in trauma patients and intra operative conditions has been widely discussed. The ratio of components to be used, trigger for initiating a massive transfusion protocol, the clinical and laboratory parameters that monitor the outcomes of the transfusion are still widely debated.

Rh incompatible transfusions in Rh negative patients are carried out in various emergency/elective situations mainly due to comparative shortage of the respective Rh negative Packed Red cell (PRBC) components and has been documented as few sporadic case reports.

Case Report: This is a case of massive transfusion which necessitated a switch over to Rh positive cells in an Rh negative patient who showed no detectable levels of anti D activity as on post op day 15.

The patient was a 65 year old male who presented to our hospital and diagnosed of left sided renal cell carcinoma with IVC infiltration and para aortic lymph node involvement with no co morbidities, posted for left nephrectomy, IVC thrombectomy and para aortic lymphadenectomy with an unremarkable pre-operative work up. His pre transfusion testing showed an unambiguous O group and a negative antibody screen.

We had cross matched and reserved 10 units of O negative PRBCs with a backup of additional 5 units reserved for this patient. The operation started with a pre-operative request for 10 units of PRBCs.

Intra operatively he developed a massive bleeding for which a massive transfusion protocol was activated and patient received 12 bags of O negative PRBCs, 24 bags of FFPs and 12 bags of Random Donor Platelets. The bleeding continued and as a result of the shortage of O negative blood units the patient had to be switched over to 12 O positive PRBCs. The post op period was unremarkable except for an additional 2 O positive PRBC transfusions. His coagulation parameters were only slightly deranged which got corrected without additional therapeutic intervention. Serial Coomb's tests were negative as on post-operative day 15.

Conclusion: Rh D switching in D negative patients who are not sensitized previously can be done cautiously with no serious adverse immunohematological events in desperate cases as in the case presented and would be a fruitful exercise as a life saving measure in such situations.

Blood donor deferral pattern in a tertiary care hospital of North India

Hitish Narang, Hardeep Kaur

SPS Hospitals, Ludhiana, Punjab, India

Background: Safe and adequate supply of blood and blood components is essential for the patients without compromising the donor safety.

Aims: To determine the reasons for donor deferral in a tertiary care hospital of North India.

Materials and Methods: The present study was conducted at SPS Apollo Hospitals, Ludhiana from April 2010 to March 2015. The records of deferred donors were analyzed retrospectively.

Results: There were 22148 blood donors during April 2010 to March 2015. Total 932 (4.2%) donors were deferred for various reasons as per NABH Guidelines. A large proportion (91.7%) of the deferred donors was replacement donors. Only 0.6% of the deferred donors were permanently deferred because of past history of cardiac ischemic disease or myocardial infarction. Anemia was the one major cause of temporary deferral, constituting 43.8% of all deferrals. Other medical causes constituted 38.5%, surgical causes 3.3% and others (like Vein not prominent for donation, previous blood donation <3 months ago etc) 14.4%. The medical causes included skin allergies, fever, infections, hypertension at the time of blood donation, alcohol/drug abuse, tattoos, H/o abortion/delivery/breastfeeding etc) and surgical causes included history of minor/major surgery.

Conclusion: The blood wastage rate can be reduced by properly screening the blood donors and deferring them after counseling for future blood donations. This small step can be beneficial for blood donors as well as blood centers, which can reduce the wastage of blood and its components and also reduce the adverse transfusion reaction rate amongst the patients.

Auto anti-C antibodies in patient of primary sclerosing cholangitis with aiha-A rare presentation

A Maheshwari, S Gupta, P Rajani, M Bajpai

Introduction: Pre-transfusion testing is a vital link to enhance patient's safety, which may be affected by autoantibodies. They may be detected during pre transfusion testing in blood bank. Primary sclerosing cholangitis is rarely associated with AIHA and the presence of specific antibodies has not been reported previously. We present a unique case report of Primary sclerosing cholangitis associated with AIHA implicating auto anti C antibodies.

Case Report: A 17 years old girl was admitted to our hospital with Primary sclerosing cholangitis along with AIHA. She had no history of blood transfusion, pregnancy or transplantation. The patient's blood grouping was O Rh D positive. Direct antiglobulin test was positive. Auto control was positive in AHG phase, confirming it as a case of warm reactive AIHA. Further testing was done to identify the autoantibody.

On three cell antibody screening and identification panel possibility of Anti-Rh-C antibody was suspected. Patient's Rh phenotype was 'C' positive (CDE/ce //++/+). For confirmation 3 samples of 'C' positive red cells and 3 samples of 'C' negative red cells were tested against the patient's serum. The 'C' negative cells were compatible while 'C' positive cell were incompatible with the patient's serum, confirming the antibody specificity as anti Rh-C. For further confirmation we performed serial alloadsorption with 'C' positive red cell. Adsorbed patient's serum showed no agglutination in the cell panels. Furthermore before alloadsorption C+ allogeneic red cells were DAT negative (control) and after allo-adsorption they became DAT positive thus confirming adsorption of anti-C antibodies. It confirmed the presence of auto anti 'C' antibody of warm type reacting at AHG phase only. As patient's haemoglobin was continuously falling and patient required transfusion of red cells, one 'C' antigen negative and crossmatch compatible unit was transfused with no untoward reactions. Meanwhile patient was given treatment with immunosuppressants. Patient's haemoglobin level and general condition showed improvement.

Conclusions: This unique case report shows Primary sclerosing cholangitis associated with AIHA due to auto anti 'C' antibody. Usually warm AIHA presents with pan reactive pattern and it is difficult to find compatible blood. In this rare case where we could find specific antibody, efforts should always be made in all cases of AIHA to identify the specificity of autoantibody whenever possible.

Study of North Indian blood donors: Attitude, obstacles and motivation towards donation

Tenzin Saldon, Garima Khanna, Balraj Shawami, Manoj Kumar, Shalini Gupta, Venencia Albert, Arulselvi Subramanian

Department of Blood Bank, Jai Prakash Narayan Apex Trauma Centre, All India Institute of Medical sciences, New Delhi, India

Introduction: Continuous efforts are required to motivate people to donate blood, to urge donors to keep on donating blood on a regular basis andto inspire non-donors to start donating blood for sustaining self-sufficiency and matching the ever increasing clinical demand for various blood components. Therefore this study was done to determine the knowledge, attitude, reasons for not donating blood, misconceptions and motivations towards blood donation among people of New Delhi.

Aim: The aim of this study is to access blood donation among people by analysing their perception and knowledge towards the cause through a questionnaire-based survey. This study also evaluates the reasons of donor deferral in a hospital-based blood bank in India. This can help blood donation centres to attract donors by developing appropriate policies and plans in future.

Materials and Methods: The study was carried out in New Delhi, India. This prospective study was done on group of 135 people during one month period. Blood Donors who visited our blood bank with age >18 years, eligible for blood donation were included in our study. Data collection was done with the help of forms given them. The form was divided into three parts and had fourteen questions regarding obstacles, misconceptions and motivations concerning blood donation. 27 females and 108 males were assessed. The respondents were briefed on the aims of the study and ways to fill the questionnaire. They were also ensured about confidentiality in this entire process. Statistics was then applied to the data and presented in forms of bar graphs for easy interpretation.

Results: The mean of age of donor population was 29.3 yrs (males: 28.9 females: 30.5) Males had higher blood donation knowledge level as compared to females in the study. 53% of males were motivated to give blood during an emergency situation of someone close while 36.5% females were motivated by story of someone being saved from blood transfusion. 28.5% males were influenced by friends. 34% girls were afraid of needles and 30%males are afraid of unknown. The main obstacle was Poor information. A message which makes one think had the highest average for a strong campaign (Females: 4.3 males: 3.8). For promoting blood donation (Average 3.9) males found television and radio programmes most effective while (Average 4.2) females found personally talking with people who have already donated blood effective. Incentives like getting free blood tests and educational credits were found to lure people. Having Proper information, being aware about the technicalities of the process and convenient place/time of donation encouraged people towards giving blood.

Conclusion: It is necessary to follow strict donor selection criteria to make blood donation safe and win the trust of future donors. The entire blood bank staff should share the responsibility of winning the confidence of donors and making blood donation a safe and pleasurable experience. The inputs from people can be useful in implementing relevant donor recruitment and to introduce strategies for maintaining an adequate and safe blood supply.

An audit of single donor platelet apheresis: A tertiary care hospital experience

Sudhir Vujhini

Nizam's Institute of Medical Sciences, Hyderabad, Telangana, India

Background: Platelet transfusions are indicated in patients who are bleeding or at increased risk of spontaneous bleeding. Platelets are currently obtained either by fractionation of whole blood or platelet apheresis. SDP has numerous advantages over RDP decreased risk of TTI's, decreased risk of bacterial contamination, decreased risk of alloimmunisation due to reduced donor exposure, and higher platelet count yield.

Aim: Audit of single donor plateletpheresis (SDP) was done to assess the utilisation of SDP, demographic profile of the donors, deferral cause of the donors, discarding details of the SDP units.

Materials and Methods: This study of single donor plateletpheresis audit was carried out retrospectively from January 2014 to May 2015 (1 year 5 months) and the data was collected from the records in the Department of Transfusion Medicine, Nizam's Institute of Medical Sciences, Hyderabad, Telangana State, India.

Patient's demographics and ABO and Rh (D) group specific or non-group specific transfusions were also assessed.

Results: Total of 124 SDP procedures (out of 165 donors) were done. All the donors were males and replacement donors. SDP accounts about 0.51% of total blood units collected and 0.88% of total platelets prepared. Most common age group of the donors was 26-30 years (46.67), followed by 18-25 years (29.03). Most common platelet count range of the donors was 2.6-3.0 lakhs/cu mm (36.29%). 41/165 (24.84%) donors were deferred. The most common cause for deferral was low platelet count. Group-specific utilization was 92.72%. Utilization of SDP units was more frequent by medical oncology department (80.64%). SDP discard rate due to expiry was 1.61%. Most common adverse effect noted was peri-oral paresthesia 56.66%.

Conclusion: Plateletpheresis is a safe procedure when proper selection criteria is employed and can be tolerated by majority of the donors as no major adverse reactions were noted. Donors who were deferred for temporary causes can be followed and repeated tests done to bring them back into the donor pool. Discarding of the SDP units due to expiry can be avoided when the requirement of SDP is correctly assessed by the clinician and a discussion with blood bank medical officer.

Trends in seroprevalance of mandatory viral markers among blood donors at tertiary care hospital

Kusum K Thakur, Kanchan Bhardwaj, Rajni Bassi, Kanika Taneja

Department of Transfusion Medicine, Government Medical College, Patiala, Punjab, India

Background: Safe blood is need of hour to prevent spread of infectious diseases like HIV, HBV & HCV. Majority of problem are due to prevalence of asymptomatic carriers in the society as well as window period of infection. Blood donors are healthiest population and their screening for HIV, HBV & HCV will reflect the true prevalence of these infections in a population.

Aims and Objectives: To know sero-prevalence of mandatory viral markers among blood donors and to compare the results in voluntary and replacement donors.

Materials and Methods: All blood units received (88502 units) during January 2010 to December 2014, were tested for HIV, HBsAg and HCV by ELISA method approved by NACO. Retrospective statistical analysis of data done and trends and co-infection were noted and compared with other similar studies. Data collection and analysis was achieved by compiling the data on Microsoft excel 2007 computing program. Simple Statistical application was used to calculate the percentage prevalence.

Observations: Among 88502 blood donors, 76910 (87%) were Voluntary and 11592 (13%) were replacement donors. Overall seroprevelance was 1.5% and mean prevalence rate of HIV, HBV & HCV were 0.07 %, 0.69% & 0.76 %. Mean positivity in voluntary and replacement donors in HIV, HBV, HCV were compared and it was more in replacement donors and was statistically significant with Chi-squire = 19.9; p value = 0.01. Co-infection was seen in replacement donors only.

Conclusion: Seroprevalence of HIV, HBV and HCV were less in voluntary donors as compared to replacement donors. Co-infection was also observed in three donors and all were replacement donors. The present study concludes that motivating voluntary blood donors by conducting voluntary blood donation camp is the most effective way of ensuring adequate supplies of safe blood on a continuing basis.

Improvement of response rate of donors for postdonation counseling

Kusum K Thakur, Shikha Aggarwal, C Sonima, Jaspreet Kaur, Kanchan Bhardwaj

Department of Transfusion Medicine, Government Medical College, Patiala, Punjab, India

Introduction: Counseling is an essential part of quality system in blood donor management. Prevention of Transfusion Transmissible Infections (TTIs) depends upon proper pre-donation counseling, selection of donors and proper screening tests done. If found positive for any TTIs, bags are discarded and donors are called back to department for post test counseling. Proper follow up of donors, with extended services of BTS in form of referral to ICTC/special clinics, increases the blood safety as there will be self deferral by them in future.

Aims and Objectives: To know improvement in response rate of donors for post donation counseling.

Materials and Methods: A retrospective analysis of records of donors for one year from September 2013 to August 2014 was done. Donors who had given consent and who were positive after screening tests, were called for further follow up.

Observations: Total number of donors during one year 2013-2014, were 17964. Total positive for TTIs after screening tests were 299 (1.6%) as compared to 237 (1.4%) in 2011-2012 but only 134 (49%) reported for post donation counseling as compared to 82 (35%) in 2011-2012 and were sent to special clinics for further management with advise that if confirmed positive should self deferral themselves for future blood donations.

Conclusion: Response rate of donors increased from 35% in year 2011-12 to 49% in 2013-14 which shows that counseling methods have improved in our center with more faith of donors in our counseling services.

Figure 1: Trends in response rate of donors reactive for viral markers

Age and gender distribution of voluntary blood donors

S Usha, D Deepa, S Shoganraj

Tamil Nadu Dr. MGR Medical University, Chennai, Tamil Nadu, India

Background: According to 2012 WHO report, India is facing a blood shortage of 3 million units. This problem could be addressed if 2 % more Indians donated blood. Demographic information of blood donors is important for formulating and monitoring recruitment strategies. As per 2008 WHO statistics, most of the voluntary unpaid donors in India, belonged to 18-24 yrs age group (53%). Only 6% of blood donors were women. We analyzed the age and gender distribution of voluntary blood donors at our centre.

Materials and Methods: Data was collected retrospectively over a 10-month period (Nov '13 to August '14) from Donor Register and the results were analyzed in terms of age and gender distribution.

Results: Among 1773 voluntary blood donors, 967 (54.54%) donors belonged to 18-24 yrs age group (highest); 738 (41.62%) in the 25-44 yrs age group and 68 (3.83%) in 45-65 yrs age group (lowest) 1709 (97.10) % were male donors and only 64 (3.60 %) were female donors.

Discussion: This study data was similar when compared to 2008 National statistics 53% (18- 24 years), 29% (25-44 years) and 19% (45-60 years) respectively, but the donation by older people is less. Similarly, blood donation by females is lower (3% compared to 6%).

Conclusion: In our study, voluntary blood donation was highest among younger people. If these donors are continuously motivated, the percentage of voluntary blood donation can be increased among older people also. Percentage of female donors is very low, if in-depth analysis for reasons behind such under performance are found out, there is a definite possibility for increase in the percentage of voluntary blood donation among female donors.

Seven year study of seroprevalence and trend of transfusion transmitted infections among blood donors in a superspeciality hospital of Delhi

Reeta Rai, Manoj Rawat, Vipin Kumar, Manjeet Pathak

Introduction/Background: Blood is a precious and scarce life saving resource but transfusion can cause life threatening infection if not tested properly. Quality and safety of blood always remains a concern for healthcare professionals.

Aim: To assess the serological prevalence of HIV, HBV, HCV, Syphilis and Malaria and also compare the yearly trend of seropositivity among blood donors in a Superspeciality Hospital of Delhi.

Materials and Methods: The study was conducted to determine seroprevalence of transfusion transmitted infection among 37,406 healthy blood donors for a period of 7 years (Jan 2008-Dec 2014). The blood donotion record of last 7 years was reviewed. Donors were selected by strictly following donor selection criteria. It included both voluntary and replacement donors. Screening for viral markers was done using Enhanced Chemiluminescence Immuno assay (Vitros Eci). Syphilis was screened by RPR method, while malarial parasite screened using malaria antigen test.

Results: A total 37,406 donors were screened during this period. There were 32,226 (86.15%) replacement and 5180 (13.85%) voluntary Donors. Female donation was only 1441 (3.85%) that included both voluntary and replacement. Male donors both voluntary and replacement were 35965 (96.15%). Overall seropositivity rate of HBsAg, HIV, HCV Syphilis and malaria 0.92%, 0.28%, 0.81%, 0.15% and 0.008% respectively. HBV seropositivity ranged from 1.03% in 2008 to 0.88% in 2014 showing decreasing trend. HIV seropositivity also showed decreasing trend from 0.42% in 2008 to 0.28% in 2014. HCV seropositivity varied from 0.84% in 2008 to 0.99%. Syphilis seropositivity also showed decreasing trend 0.34% in 2008 to 0.11 in 2014. Prevalence rate of transfusion infections among Replacement donor is high (2.3%) as compared to Voluntary Donors (1.3%).

Conclusion: Our study shows overall seroprevalence of HBV and HIV is low and showing decreasing trend, while in HCV seroprevalence is slightly high as compared to other studies which may be due to better diagnostic kit with higher sensitivity. Seroprevalence of syphilis is low although with some fluctuation. Low seropositivity in our study could be due to strict donor selection criteria and predonation counselling. Seropositivity rate among voluntary donor is low as compared to Replacement donors. In view of this, efforts should be done for promoting voluntary donation and strict donor selection criteria must be followed to make the blood more safe.

Implementation of multi-dye nucleic acid testing for blood screening in Delhi, India

Sangeeta Pathak, Surjit Singh, Tamojit Chakraborty, Ruchi Dubey

Max Super Specialty Hospital, New Delhi, India

Background: Nucleic Acid Testing (NAT) was first introduced at Max Super Specialty blood bank as part of the blood screening system in July 2010. Upon initial NAT implementation, blood donations were screened in pools of six using the Roche cobasTaqScreen MPXv1.0 test (MPX v1 test) which requires further discrimination of a reactive donation, in order to identify the reactive viral target. In September 2014, NAT was upgraded to the newly launched Roche cobasTaqScreen MPX v2.0 test (MPX v2 test) in India, which employs real-time detection and identification of 3 viruses in a single test by using multi-dye PCR technology. This technology enables the direct detection of HIV, HBV, HCV and the internal control in separate channels thus removing the need for any additional viral discriminatory testing.

Aim: Evaluation of the MPX v2 test performance for routine blood screening.

Methods: A total of 90,396 seronegative donations were screened by NAT in minipools of six from July 2010 till Feb 2015 for HIV-1 & -2 RNA, HCV RNA and HBV DNA. Among the donations tested, a total of 77,614 donations were tested on the MPX v1 test from July 2010 till August 2014 and thereafter, 12,782 donations were tested on the MPX v2 test from September 2014 till February 2015.

Results: During the initial 4 years and 2 months of NAT implementation using the MPX v1 test, there were a total of 17 HBV NAT yield donations detected (NAT reactive but seronegative cases) and 3 HCV NAT yield and 2 HIV-1 NAT yield cases detected. This would generate a NAT yield rate of 1:4566 for HBV, 1:25871 for HCV and 1:38,807 for HIV-1. With the advent of MPX v2 test with increased HBV analytical sensitivity, in the last 7 months, we have detected 3 HBV NAT yield cases with a rate HBV yield at 1:4,261. In an earlier study, data presented for the first 18 months of NAT, the HBV NAT yield reported was 1:3355, the HCV yield was 1:20,131 and the HIV-1 yield was 1:10,065. We now observe that these rates based on cumulative data on the MPX v1 test seem to have decreased. Viral load testing performed on the HBV NAT cases showed that the 13 of the 17 cases had very low level titres of <20 IU/ml, three with viral titres of 27-33 IU/ml and only one with a viral load of 4541 IU/ml.

Conclusion: The implementation of NAT at our blood bank in addition to screening with serology continues to circumvent the risk of infected blood units being transfused. We will continue to monitor the rate of the HBV NAT yield cases to understand the effect of the increased sensitivity with the MPX v2 test, and the corresponding slight increase of HBV NAT yield rate. In general, the use of the MPX v2 test with real-time viral discrimination allows for faster identification of the reactive donation and prompt notification of the donors for counselling.

Haemovigiltm: A novel system to prevent bedside phlebotomy and transfusion errors due to patient mis-identification

Sangeeta Pathak

Max Super Specialty Hospital, New Delhi, India

Background: Over the last 2-3 decades, sensitive diagnostic tests has resulted in a drastic reduction in transfusion-transmitted infections (TTI). In fact, Wrong Blood in Tube (WBIT) and incorrect patient identification results in much higher morbidity and mortality compared to TTI1. [2] The first years of Serious Hazards of Transfusion (SHOT) also confirmed that patients were more at risk of a wrong transfusion than of any other transfusion reaction. [3]

Aims: This institute carried out the first such study in the world using HaemovigilTM transfusion safety system. HaemovigilTM is designed to prevent errors in the entire transfusion process from patient's blood sample collection, allotting and reserving units of blood for the patient in the blood bank to a final bedside confirmation that the patient is the same, from whom the blood sample was drawn, before being able to start any transfusion. The data collected during the study period of 4 months was compared to the data collected over the last 3 years to see if the intervention could have prevented any errors. The aims of the study were to identify and prevent.

WBIT errors where in the sample of a wrong patient is sent to the blood bank for crossmatch

Issue of incorrect reserved unit where in the blood bank issues a component unit corssmatched and reserved for some another recipient

Incorrect or no final identification of patient where in the final bedside pretransfusion identity confirmation of patients does not take place

First hand feedback from nursing staff about the ease of use of HaemovigilTM.

Retrospective data of the last 3 years (2011-2013) was analyzed to assess the number of errors of sampling (improperly labeled samples, samples without patient information and WBIT). Over a period of 4 months (September 2014 to December 2014), we implemented HaemovigilTM in the transfusion process. HaemovigilTM comprises of:

Haemovigil Transfusion Wristbands - Having a unique 6 digit number and 4 peel off labels with encrypted codes

Haemovigil Software: Which decrypts the code and geneartes the same 6 digit number as present on the wrsitband from which the label was peeled off

Haemovigil Digital Transporter - A thermal insulated box with a digital elctronic lock in which the unit of blood is transported and the box can be opened by entering the unique 6 digit number on the Haemovigil Transfusion Wristbands.

Results: Analysis of retrospective data of the 3 years was as follows

This error rate (0.28%) was in line with most of the published data. [4] We did not have data for incorrect identification of patient prior to transfusion or transfusion of blood to a wrong patient. However, there were at least 2 incidences in the period of 3 years where the patient was moved to a different ward/bed and the nursing staff was unaware that this (near miss event).

In this study, 800 patients (in few wards) were given the Haemovigil Transfusion Wristbands at the time of admission. The number of units transfused (irrespective of component) to these 800 patients is given in the table below.

Analysis of data duringthe study was as follows

1 unit of blood was not released by the Haemovigil software as the name of the patient was different during unit release request than from the requisition form sent earlier. The Haemovigil software works on the basis that the patient name and number on the wristband should match before reserved units are released.

1 incidence of the Haemovigil box not being opened as the patient's wristband was taken off during transfer from one ward to another. Since Haemovigil can only be opened by entering the unique number on the patient's wristband, any change in patient bed/ward can be detected by the system.

1 incident where an untrained staff wasn't able to open the box. Haemovigil ensured that an untrained staff wasn't able to start the transfusion process. In the absence of this system, untrained staff would still have gone ahead with the transfusion process.

This pilot study was carried out to study the viability and ease of use the Haemovigil apart from preventing errors. During the study period, there wasn't any case of sample error. The implementation of Haemovigil made it easier for the phlebotomist to label blood sample at the patient bedside during collection of blood. Being in line with their workflow and they found the system easy to implement. There were 3 errors in the study period that could be termed as near miss events that were detected and prevented the Haemovigil. Training of staff has its pitfalls as attrition and human errors cannot be prevented. On the contrary, implementation of the Haemovigil system ensures that all protocols are followed. Haemovigil is cost effective technology driven intervention to remove human errors during specimen collection, blood unit allocation in the blood bank and final patient identification prior to starting transfusion. It is an affordable and easily implementable system that ensures adherence to a specimen-labeling policy and a final check of the patient identity. Any variation in the protocol ensures that the unit of blood cannot be taken out from the box and this will go a long in avoiding transfusion errors.

Further Reading

1. Varey A, Tinegate H, Robertson J, Watson D, Iqbal A. Factors predisposing to wrong blood in tube incidents: a year's experience in the North East of England. Transfus Med 2013;23:321-5.

2. Dzik WH, Murphy MF, Andreu G, Heddle N, Hogman C, Kekomaki R, et al. An international study of the performance of sample collection from patients. Vox Sang 2003;85:40-7.

3. Bolton-Maggs PH, Cohen H. Serious hazards of transfusion (SHOT) haemovigilance and progress is improving transfusion safety. Br J Haematol 2013;163:303-14.

4. Grimm E, Friedberg RC, Wilkinson DS, AuBuchon JP, Souers RJ, Lehman CM. Blood bank safety practices: mislabeled samples and wrong blood in tube - a Q-probes analysis of 122 clinical laboratories. Arch Pathol Lab Med 2010;134:1108-15.

Reducing blood transfusion requirement in bilateral total knee replacements

Sadhana Mangwana, Ashok Goel, Ritu Gupta

Blood Transfusion Services, Sri Balaji Action Medical Institute,

New Delhi, India

Introduction: Total Knee Replacement is a commonly performed procedure. In the Asian setup, patients usually come late and there may be requirement for both knees to be replaced at the same sitting. This may increase the requirement for blood transfusion. Our aim is to reduce blood transfusion rate in a bilateral total knee replacement.

Methods: We present a prospective series of 75 consecutive bilateral total knee replacements from April 2013 to March 2015. There were 18 males and 57 females. The mean age was 64 (range 46-78). Exclusion criteria includes pre-operative haemoglobin less than 11 gm% and patients deemed unsuitable for bilateral total knee replacement at the same sitting. Our criteria for transfusion is post-operative haemoglobin less than 8.0 gm% or clinically symptomatic. A standard protocol is used. If the patient is on Aspirin and/or Clopidogeral the drugs are stopped five days prior to surgery. An epidural anaethesia is given to all patients. The posterior capsule is infiltrated with a mixture of 60ml of 0.2% Rupivacaine and 2 ml of 1:100,000 Adrenaline. The tourniquet is released after the cement has set, the wound is thoroughly washed with normal saline and all bleeding points diathermized. No drains are used and a compression bandage is applied. The compression is left in place for 48 hours. Low molecular weight Heparin is given in the post-operative period and this is switched to Aspirin on discharge. The post-operative haemoglobin is measured and documented on the first post-operative day.

Results: In this study the pre-operative haemoglobin was 11 to 17 gm% (mean Hb: 12.5 gm%). The post-operative haemoglobin was 7.5 to 14.1 gm% (mean Hb: 9.9 gm%). Our blood transfusion rate in the 75 consecutive bilateral total knee replacement was 6.67%. Out of the 5 patients requiring blood transfusion, 4 patients were on pre-operative Clopidogrel and in one patient no relationship to any cause could be ascertained.

Discussion and Conclusion: The blood transfusion requirement, even in a bilateral total knee replacement, may be reduced considerably by simple measures. These include: Epidural anaesthesia, posterior capsule infiltration, no drains, thorough wash and haemostasis before closure and post-operative compression bandage for 48 hours. There may be a need for blood transfusion in patients who have been on pre-operative Clopidogrel. These measures not only reduce the need for handling blood products but also prove cost-effective by saving man hours and money in component preparation, cross matching and reserving blood for the patients. It prevents blood transfusion reactions; immediate and delayed (Transmission of transfusion transmitted infections), and associated problems. It also conserves the blood for other emergencies.

Effect of new algorithm in nucleic acid testing (enact study)

Aseem Kumar Tiwari, Ravi C Dara, Dinesh Arora, Rawat Ganesh, Raina Vimarsh

Department of Transfusion Medicine, Medanta, Gurgaon, India

Introduction: In India, many blood centers using Nucleic acid testing (NAT) have published their experience of NAT yield based on the "Algorithm A" where, if the individual donor sample was reactive on a multiplex NAT (ID-NAT) after negative serological screening test, the donation was subsequently tested with discriminatory NAT. We introduced a new algorithm based on replicate testing and compared the results with the previous algorithm.

Materials and Methods: Results of NAT using the algorithm A from January 2011 until June 2012 and the algorithm B from July 2012 until August 2014 were analysed in large tertiary care hospital of India. Percentage of concordant positives (serology and NAT reactive), concordant negatives (serology and NAT non-reactive), seroyield (serology reactive and NAT non-reactive) and discriminated NAT yield (serology non-reactive and NAT reactive) were compared between the two algorithms. Sample to cut offs (S/CO) of the donations analyzed as repeatable reactive (RR) and its subsets were compared.

Results: 88583 blood donor samples tested during the study period (31844 with algorithm A and 56739 with algorithm B). Among 1037 reactive donations, p value of serology and NAT concordant (252 with algorithm A and 446 with algorithm B) and discordant donor samples (42 seroyield with algorithm A and 68 with algorithm B and 147 NAT initial reactive with algorithm A and 82 with algorithm B) were highly significant (p = 0.0001). NAT false positivity came down from 95.2% in algorithm A to 73.1% in algorithm B (p = 0.0001). Discriminated yield (DY) rate went up from 4.7%in algorithm A to 21.9% in algorithm B (p = 0.001). Significant (p < 0.001) difference in S/CO between "Non Repeatable Reactive (NRR) and DY" and "DY and Non Discriminated Yield (NDY)" was seen.

Conclusion: Study recommends replicate testing strategy in ID- NAT testing that will increase the NAT yield rate and will eliminate the unnecessary discriminatory tests.

Plasma and its derivatives: Their effect on conventional coagulation screening tests

Manish Raturi, Shamee Shastry, Mohandoss Murugesan, B Poornima Baliga, P Kalyana Chakravarthy

Department of Immunohematology and Blood Transfusion, Kasturba Medical College, Manipal University, Manipal, Karnataka, India

Background: Conventional coagulation screening parameters are most often used to predict bleeding in various clinical situations. The present study aims to observe the effect of plasma and its derivatives on the conventional laboratory coagulation test results and their hemostatic benefit in the patients.

Materials and Methods: We have done prospective analysis of all the requests for plasma in a tertiary care referral center. Patients' PT, INR, aPTT, INR, platelet count and fibrinogen levels within 12 hours preceding the requests and 24 hours post transfusion were noted to assess the level of correction. The magnitude of improvement in INR was determined using the formula by Holland and Brooks. Both the changes in laboratory parameters as well as the clinical cessation of bleeding was considered to evaluate the therapeutic efficacy.

Results: Among 2082 episodes, 4991 units of plasma were utilized at an average of 5 units per patient and in 74 episodes of cryoprecipitate transfusions, 419 units were utilized at an average of 8 units per patient transfused. The mean change in INR following plasma transfusion is 8.9% of pre-transfusion INR and thus considered as significant change. Clinical improvement following FFP transfusion was greater in the group with higher INR. The mean increment of fibrinogen was 79.8 mg/dL. A weak negative correlation was observed between the mean pre-transfusion fibrinogen level and the mean dose of cryoprecipitate transfused.

Conclusion: Transfusion of plasma is not effective in patients with mildly deranged coagulation laboratory values and decision to do so shall be reconsidered by the clinicians.

Immunohematological problems in liver transplant patients: Their types and resolution

Aseem K Tiwari, Ravi C Dara, Dinesh Arora, Subhasis Mitra, Vimarsh Raina

Department of Transfusion Medicine, Medanta, Gurgaon, India

Background: Transplantation is now a common procedure for replacing the dysfunctional liver. Improvements in surgical techniques, anesthetic management, organ conservation, better knowledge of hemostatic agents have contributed to reduction of transfusion of various blood components. However, great variability in transfusion rates between centers have been published, ranging between 0 and more than 100 RBC units transfused during the liver transplants procedure. Red cell allo-immunization adds to this as the presence of red cell alloantibodies in patients awaiting a liver transplant may cause delay or unavailability of compatible red blood cell (RBC) units creating pressure over caregivers. In this situation compatible blood units can be provided by well-equipped immunohematology laboratory with expertise in resolving serological problems. In this case series, we present various types of Immunohematological problems and our approach in resolving these complicated cases in liver transplant setting.

Case 1 was a 49 year old male diagnosed with End Stage Liver Disease (ESLD) scheduled for liver transplant. Blood Transfusion Services (BTS) received request for blood grouping and antibody screening and providing compatible red cell units. On antibody screening and identification initially two allo-antibodies (anti-c and anti-E) were identified but cross-match was found incompatible with corresponding antigen negative (c−, E−) blood units suggesting presence of third antibody. This antibody (anti Fyb) resolved by using select cells. Patient underwent liver transplant with eight compatible units and continues to be alright with normal liver function tests and normal hemoglobin (10.2 gm %).

Case 2 was again a patient with ESLD with diabetes posted for liver transplant with multiple irregular allo-antibodies with the history of severe hemolytic transfusion reactions. On antibody identification Anti C, Jka and S were identified with the titer of 64, 128 and 16 respectively. Providing compatible units for liver transplant would have meant screening 917 units, number higher than the inventory; thus pre-operative plasma exchange was done to reduce titer of alloantibodies and antigen negative fully compatible units along with some partially matched units were used in the surgery for transplant successfully.

Case 3 was also a patient with ESLD for liver transplant in distant city with multiple antibodies (Anti C, Anti e and Anti K). To find out these three antigen negative blood unit would have meant screening 1432 units. Therefore search was done in rare donor registry and inventories both, for corresponding antigen negative donors. Two compatible units were provided to this patient one from donor registry and one from RBC inventory.

Conclusion: Immunohematological problems can complicate liver transplant procedures. However, there are several approaches to over these complications and allowing successful patient and transplant outcomes.

A simple PCR assay for identification of RHD variants

Swati Kulkarni, G Vidya, K Ghosh

National Institute of Immunohaematology, Mumbai, Maharashtra, India

Background: The Rh blood group discrepancies may arise when an individual is a variant of D antigen. These variants are identified as RhD negative or positive depending on the reagents in use and techniques used in different laboratories. In apparently D negative individuals, D variants may remain undetected due to the limitations of serology. These variants are of clinical importance and should be identified as they may produce anti-D when transfused with normal D positive red cells. Molecular RHD typing has been shown in literature to overcome the limitations of serologic methods.

Aims and Objectives: To standardize simple PCR based technique for identification of RHD variants.

Materials and Methods: A total of 700 RhD negative samples were taken for the study. A PCR based on the allele specific amplification of the the 3' noncoding region of exon 10 of the RHD gene and exon 7 specific internal control was standardised. Samples showing presence of exon 10 of RHD gene were tested with ALBAClone partial D typing kit and the most probable Rh phenotype determined using anti-C, c, D, E, and e antisera.

Results: RHD specific PCR amplification was observed in 28 samples (4%). All these samples showed presence of "C" antigen and were found to be D variants by partial D typing kit.

Conclusion: A simple RHD PCR specific for exon 10 of the RHD gene was standardized that could be used as a screening method for identification of D variants.

Database of extensively phenotyped regular blood donors for transfusion support of thalassemic patients

Swati Kulkarni, Bhavika Choudhary, K Ghosh

National Institute of Immunohaematology, Mumbai, Maharashtra, India

Thalassemic patients requiring chronic transfusion support are at risk of alloimmunisation because of disparity between donor and recipient antigen profile. The presence of RBC alloantibodies creates the potential for serologic incompatibility, makes the selection of appropriate units for future transfusion more difficult and presents risk of hemolytic transfusion reaction. Matching for the critical antigens of Rh, Kell, Kidd and Duffy blood group systems has been shown in literature to dramatically minimize alloimmunisation.

Aim: To create a database of regular blood donors (phenotyped for common antigens of Rh, Kell, Kidd and Duffy blood group systems) for providing extended antigen matched RBCs for transfusion support of thalassemic patients.

Materials and Methods: Blood samples from seventy multitransfused beta-thalassemia patients were tested for antigens or genes of Rh, Kell, Kidd and Duffy blood group system by hemagglutination and PCR technique. Two hundred and forty five 'O' group regular blood donors were serologically phenotyped for the above antigens.

Results: Out of seventy beta-thalassemic patients tested, the genotyping was concordant with the serological red cell phenotype in only fourteen (20%) cases for five antithetical antigen pairs belonging to four blood group systems. Based on partial matching of donors and patients for Rh antigens (C, c, E, e) we identified 97 R1R1 donors for 37 thalassemic patients, 66 R1r donors for 15 patients, 25 donors R1R2 for 11 patients, five R0r donors for four patients and 35 rr donors for three patients. A total of 153 regular donors matching perfectly for D, C, c, E, e, Fya, Fyb, K, k, Jka, Jkb antigens were identified for 70 thalassemic patients.

Conclusion: Blood group genotyping enabled determination of blood group when serology failed due to presence of transfused red cells. Database of regular blood donors phenotyped for Rh, Duffy, Kell, Kidd blood group system antigens was created which will help in providing extended antigen matched RBCs for thalassemic patients.

Noninvasive fetal RHD genotyping using cell-free fetal DNA from maternal plasma

Disha Parchure, Swati Kulkarni, K Ghosh

National Institute of Immunohaematology, Mumbai, Maharashtra, India

Background: Prenatal fetal RHD typing traditionally was carried out using invasive techniques like Chorionic villus sampling and amniocentesis. These invasive methods carry a risk of transplacental haemorrhage and pregnancy loss. Currently, determination of fetal RHD status from maternal plasma using cell-free fetal DNA has become an exciting tool for the management of RhD negative pregnant women.

Objective: To standardize real time PCR for noninvasive fetal RHD typing using maternal plasma.

Materials and Methods: Noninvasive fetal RHD genotyping was performed on blood samples of twenty five nonimmunised and five alloimmunised RhD negative women (second trimester). Taqman probes were used for real time PCR to detect exon 4, 5 and 10 of the RHD gene. SRY gene and CCR5 gene probes were used as controls. The PCR results were compared with the RhD status of babies after delivery.

Observations: Real time PCR method for noninvasive fetal RHD genotyping was standardized. There is complete concordance between results obtained by noninvasive fetal RHD typing and serological typing on cord blood sample.

Conclusion: Noninvasive testing for RHD gene by Real time PCR provides a safer alternative for the management of RhD negative pregnant women who are isoimmunised and will prove cost-effective with a substantial reduction of costs of overall immunoprophylaxis when the fetus is RhD negative.

Key words: Cell-free fetal DNA, noninvasive fetal RHD genotyping

Comparative study of two cell separators used for peripheral blood stem cell collection

Vikas Hegde, Rasika D Setia, H Shavangi, Meenu Kapoor, Shalu Bajaj, Bir Singh

Background: Amicus and Cobe spectra are the two versatile machines used commonly in transfusion medicine setup. Various studies across the globe have compared them with respect to the plateletpheresis. We have compared these machines with respect to various aspects of peripheral blood stem cell collection (PBSC).

Aims: To analyze these machines based on the donor related parameters such as donor comfort, donor reactions, time taken, volume of the blood processed during the procedure, product related parameters like volume of the product, dose of CD34+ cells, contamination by RBCs, platelets and Collection efficiency (CE%).

Materials and Methods: We retrospectively analyzed the data of successful autologous and alleogenic PBSC procedures for 5 consecutive years. The machines were run on default parameters. The study population was divided into 2 groups- one for each machine. Data of both the groups were retrieved with respect the aims mentioned above. Unpaired sample T-Test applied to compare the parameters of each machine.

Results: Collection efficiency of both the machines was comparable (amicus = 64.4% v/s cobe = 55.8%, p = 0.422). Amicus took more time to process the product (near significant p = 0.06). Volume of the product was significantly higher on Cobe for the same number of CD34+ cells (317 ml v/s 216 ml p = 0.00). Cobe collected significantly high number of platelets/΅l than Amicus in the product (1677 v/s 603 p = 0.00).There was no significant difference in the procedure details such as volume of blood processed, no. of donor reactions (citrate toxicity) and product parameters such as Hb%, TLC, MNC and total CD34+ count.

Conclusions: Though the machines were comparable in many aspects, Cobe may be disadvantageous in the sense that product volume being more, more DMSO used for cryopreservation and more platelet extracted may compromise a patient autologous donor.

Pure red cell aplasia following abo mismatched allogenic stem cell transplant

Vikas Hegde, Shiv Kumar Soni, Rasika D Setia, Meenu Kapoor, Shabih Nayyer

Background: Pure red cell aplasia (PRCA) is a potential adverse consequence of major ABO mismatched hematopoietic stem cell transplantation (HSCT), that results from recipient plasma cells continuing to produce antibody against donor ABO antigen.

Aims: The aim of the study was to know the incidence of PRCA, initial ABO titre in PRCA cases and salvage options.

Materials and Methods: We conducted the study retrospectively from the data of last 5 years. The data from 37 major ABO incompatible transplants were collected. PRCA was defined as profound peripheral blood reticulocytopenia (<10X10 9 /L) for more than 60 days with absence of erythroid precursor on bone marrow biopsy and normal myeloid, lymphoid and megakaryocytic population. ABO titration was done in all in all major ABO incompatible cases to know the initial titre and followed up until it came down to 1:32 before transplant.

Results: Five out of 35 (7%) patients who received major or bidirectional ABO mismatch incompatible HPSC transplant developed post transplant PRCA. Three patients with PRCA had blood group O and had received a major ABO incompatible transplant from blood group B (n = 2) and A (n = 1) while two cases of PRCA was observed in bidirectional ABO blood group incompatible transplant. The median pre-transplant anti donor isoagglutinin titers in patients with PRCA was 1:128. The CD 34+ cell dose was a median 4.5 x 106/Kg/L). 4 patients were successfully treated with corticosteroids. One case required additional rituximab for the recovery.

Conclusions: Our study revealed that the incidence of PRCA was 7%which is lesser than the usually reported number of 15-20%. Initial ABO titre in PRCA cases was 1:128. This helps the clinician to be cautious in choosing patients with initial titres above 128. PRCA can be successfully treated with corticosteroids and rituximab.

Comparing two hemoglobinometers used for blood donor hemoglobin estimation

Rasika D Setia, Vikas Hegde, Shalu Bajaj, KA Amit

Background: Screening of blood donors for hemoglobin (Hb%) estimation is an important aspect of donor selection. Of the various methods of hemoglobin estimation copper sulphate method is widely practiced throughout the country. However this method is being replaced by the use of hemoglobinometers (HBM) because they are more accurate, easier to handle, determines both upper and lower limits.

Aims: The aim of the study was to compare the two HBMs HEMOCUETM 201 and COMPOLAB TS with respect to accuracy, sensitivity, specificity as compared with a standard machine and the Turnaround time (TAT).

Materials and Methods: We conducted the study prospectively. The data from 100 donors were collected. Donors with HB% lower than 12.5 g% and greater than 17 g% were allowed to participate although their blood was not taken as donation. The statistical analysis was done using unpaired t-test was used to compare Hemocue, autoanalyser and Compolab readings.

Results: Of the 100 donors 10 donors had the HB% out of range (<12.5 and >17 g%). 2% had the HB % above the upper limit. The range which the instruments reported the HB% was between 7.1 to 17.5 g%. The unpaired t test showed that mean differences were significant (P = 0.000) when Compolab was compared with autoanalyser as well as the Hemocue. However there was no significant difference between Hemocue and autoanalyser (P = 0.346). As compared with the standard instrument the hemocue was more sensitive (100% v/s 90%) whereas the both had equal specificity (96%) for out of range values. Compolab had a mean TAT 2.58 sec and hemocue of 58 sec.

Conclusions: Statistical analysis revealed the superiority of Hemocue in being more accurate and sensitive to out of range values machine as compared with the Compolab indicating that one would go for the Hemocue in order to screen the blood donors. Accurate readings are essential to rule out the donors with out of the range HB% and also a more sensitive screening test is preferable to screen the donors.

Managing transfusion support in a thalassemia patient with warm antibodies

Heena Jobanputra, Jyoti Bhatt, Chandni Karia, Falguni Jani, Sanjiv Nandani

Rajkot Voluntary Blood Bank and Research Centre, Rajkot, Gujarat, India

Background: Allo-immunization is a common effect of multiple blood transfusions. Patients who are transfused with multiple units of blood, like thalassemia patients have more chances to develop allo-immunization. In this type of patients, phenotype matched blood is required.

Aim: To identify antibodies and provide safer blood in multi transfused patient to save the patient lives in critical condition.

Materials and Methods: Patient's sample was analyzed by routine blood bank workflow. Antibody screening test was positive by Solid Phase Red Cell Adherence (SPRCA) method. Antibody screening study of this sample was done by both the Conventional Test Tube (CTT) method and Column Agglutination Technique (CAT). Auto Control and Direct Anti Globulin Test (DAT) by both the methods was also done. We carried out Antigen phenotyping (advanced RCS laboratory) while antibody identification was done at reference laboratory.

Results: Patient's Antigen phenotyping result was suggestive of "cc ee"(Rh Antigen) and K-negative (Kell Antigen)". DAT and auto control were positive. The presence of any underlying allo-antibody could not be determined after multiple allo-adsorption due to high titers of masking auto antibodies.

Conclusion: Before detection of warm auto-antibodies, this thalassemia patient was transfused every 06-08 days. After Partial Phenotype Matched (PPM) and AHG compatible blood transfusion, the transfusion interval was increased up to 22 days. This was achieved with blood bank efforts as well as help of dedicated phenotype matched blood donors. We can avoid transfusion reaction if we perform phenotype matching of blood donors for patients with unidentified Antibodies.

Voluntary blood donors versus microbes: The real fact

Nishith Vachhani, Chandni Karia, Ekta Pankhaniya, Sanjiv Nandani

Rajkot Voluntary Blood Bank and Research Centre, Rajkot, Gujarat, India

Background: It is quite obvious that Blood can save lives. However, blood transfusion may lead to many complications in recipients. It is mandatory to screen all donated blood units for five transfusion transmitted diseases - HIV, hepatitis B and C, malaria and syphilis. The prevalence of these TTIs varies by nationality and geography. This group of microbes remains key factor in field of blood donation.

Aims: This study was conducted to study the prevalence of these infectious diseases among voluntary blood donors at the stand alone Rajkot Voluntary Blood Bank and Research Centre, Rajkot in Gujarat.

Methods: All blood donors were volunteers and unpaid. Donors were screened thoroughly based on the history, physical and hematological examinations before donating blood. Serological markers of HBV, HCV and HIV-1, 2 were tested by enzyme-linked immunosorbent assay (ELISA); HIV testing was done by using fourth generation ELISA kits from 2009. Syphilis was tested by rapid flocculation method. Malaria was tested using rapid immunochromatographic assay.

Results: Total 140955 voluntary blood donors (130932 male and 10023 female) were screened over a period of 07 years from January, 2008 to December, 2014. During the study period, prevalence of HIV, HBV, HCV, Malaria and syphilis were 0.15%, 0.67%, 0.11%, 0.03%, 0.10%. The prevalence of infections was 1.08% in male and 0.73% in female donors. HBV was the most common culprit in case of voluntary blood donors (0.67%). HIV, HBV and HCV prevalence rates were tends to increase age wise up to 40 years. Maximum infections were found between 31-40 years of age group (35.33%). Most of double sero reactivity is found in out-door blood donation camp (0.76%) in comparison to in-house blood collection (0.30%). Total 13 cases of mix infections were found including HIV-HBV (04 cases), HIV-syphilis (05 cases) and HBV-syphilis (04 cases).

Conclusions: This prevalence of TTI and their trends over the years were studied. A moderately fluctuating trend was observes in our study for all infections with a slight reduction in HIV and malaria infection in the recent years. Due to the initiation of government and awareness of common people, HIV and malaria is found to be least transfusion transmitted disease. Sensitive and assured donor screening technique and a stringent quality assurance system are essential to maintain blood safety along with mass awareness program of voluntary blood donor society regarding TTI is today's demand.

Seroprevalence of transfusion transmitted infections among blood donors at a tertiary care hospital blood bank in North India

Dev Raj Arya, NL Mahawar, Rashi Pachaury, Arun Bharti, DP Soni, Lokesh Sharma, Sangeeta Kumari, Pankaj Kumar Das

Department of Immunohaematology and Transfusion Medicine, S.P. Medical College and A.G. of Hospitals, Bikaner, Rajasthan, India

Background: Blood transfusion service is a sensitive issue as it is covered by 'Drug and Cosmetics Act' and has legal implications. Strict criteria are followed while selecting a donor so that proper blood free of all pathogens is available for recipient.

Objectives: To study seroprevalence of transfusion transmitted major infections (HIV, HBV, HCV, Syphilis, Malaria) among blood donors at a tertiary care government hospital blood bank in North India and to compare positivity of markers of these infections in voluntary and replacement donors.

Methodology: Present study was conducted over a period of five years (July 2010 to June 2015). Total number of donors screened during this period was 130920. Samples were tested for HIV, HBV, HCV, Syphilis, Malaria and retested if found positive by third generation ELISA tests (HIV, HBsAg, HCV). Comparative analysis was done using Chi-square for linear trend. Comparison between prevalence rates among voluntary and replacement donors was done using Chi-square tests using Excel Microsoft 2007.

Results: Of total 130920 donors, 114214 (87%) were voluntary and 16706 (13%) were replacement donors. Male donors predominated, 128781 (98.37%) male and female 2139 (1.63%). Average seropositivity of HIV, HBsAg, anti-HCV, Syphilis and Malaria was 0.10%, 1.60%, 0.18%, 0.89% and 0.04% among all donors. Significant difference (p < 0.001) in the seropositivity of HIV, HBsAg, HCV, Syphilis was seen between voluntary and replacement donors. Seroprevalence of HIV, HBsAg, HCV, Syphilis and Malaria were 0.08%, 1.56%, 0.13%, 0.71%, 0.03%, and 0.25%, 1.90% 0.51%, 2.12%, 0.04% in VBD and RBD respectively.

Conclusion: Comparing a retrospective data over a long period showed decreasing trend in seroprevalence of HIV, HBsAg, anti-HCV, Syphilis and Malaria. Results of the study reflect prevalence of these infections in the healthy population and warrant measures that should be taken to detect these infections and prevent transmission.

A case report on red cell exchange in an allo-immunized sickle cell and thalassemia double heterozygous patient posted for total hip arthroplasty and shoulder hemiarthroplasty

Ruhi Mehra, Seema Gupta, Ujwala Maheshwari

MGM Medical College and Hospital, Navi Mumbai, Maharashtra, India

Introduction: Sickle cell disease (SCD) is an inherited disorder caused by an abnormal hemoglobin (HbS). Sickle-cell β thalassemia is double heterozygous state, it is divided into two types - Sickle-cell βo thalassemia (βsβo) and sickle-cell β+ thalassemia (βsβ+).

Majority of patients with SCD receive transfusions at some point in their life to reduce complications of the disease. Patients may become allo-immunized due to repeated transfusions. Red Cell Exchange (RCE) transfusion reduces the concentration of sickle cells without increasing the hematocrit or whole blood viscosity, it can be performed manually or by erythrocytapheresis.

Here a case of sickle-cell β+ thalassemia (βsβ+) patient, alloimmunized with anti-c, posted for hip arthroplasty and shoulder hemiarthroplasty who underwent RCE transfusion is reported.

Case Report: A 36 year old male, know case of sickle-cell β+ thalassemia (βsβ+) disease presented with AVN of head of humerus. The patient had a history of AVN of femoral head 2 years back for which he was operated unsuccessfully elsewhere, hence, he was posted now for both shoulder hemiarthroplasty and total hip arthroplasty. He had received transfusions in the past whenever required. HPLC done during the pre-operative workup of the patient revealed HbS 70.2% with Hb of 11.6g/dl. RCE transfusion was required to be done before surgery. The pre-transfusion workup of the patient revealed his blood group to be AB Rh D Positive. 3 cell and11 cell antibody screening and identification panels (Diacell, Biorad, 1785, Cressier s/Morat, Switzerland) were positive for the presence of anti-c antibody reacting at 37oC. DAT and autocontrol were negative. The antigen typing of the patient revealed him to be negative for c antigen. Out of 40 donors units cross-matched with the patient sample only 5 were found to be compatible. The patient was given manual red cell exchange in which 3 units of whole blood were removed and 500 ml saline & 3 units of compatible, limited phenotypically-matched, sickle-negative, leuko-depleted PRBC's, less than 7 days old were transfused to the patient over 2 days. The HbS levels decreased post RCE and pt was operated successfully on the 3 rd day. 1 blood unit was transfused post-operatively, the pt had mild febrile reactions during transfusions which were managed successfully. The patient was stable post operatively.

Discussion: Vaso-occlusive crises occur in patients of SCD that leads to AVN of ends of long bones. In allo-immunized patients, finding compatible units can sometimes be difficult. RCE transfusion is an effective but perhaps underutilized therapy for both acute and chronic complications of SCD. It possibly reduces iron accumulation since a volume of packed cells is removed equal to the HbA containing cells that are infused. The suggested goal of the exchange is HbS less than 30% and Hb close to but not greater than 10-12g/dl.

Comparative evaluation of gel column agglutination technology systems and commercial reagent red cell panel manufactured in India for antibody screening

C Dhinesh Kumar, Dheeraj Joshi, K Muthamil Selvi

Background: The screening cells presently available and used for screening alloantibodies, are of Caucasian origin. Availability, cost and Ab indegenious to Indian groups can be missed with the Caucasian cells. Screening cells from local ethnic groups would be advantageous and increases the probability of detecting antibodies against antigens in local population.

Aims/Objective: This study was to evaluate and compare the performance of CAT Systems and their antibody screening cells from:

Indian population (Tulip)

Non-Indian origin cells.

Materials and Methods: In this comparative evaluation, CAT Systems and respective antibody screening cell panels from two different population were included. Matrix Gel System and Matrix ERYGEN-AS from India- Tulip Diagnostics and CAT System and cell panels manufactured from Non-Indian origin were used. Total 306 patient's samples from multiple transfused patients, Anemic, different pre-operative profiles and antenatal cases etc from a Tertiary care hospital were tested for antibody screening. In positive samples antibodies were further identified for specificity.

Observation: Out of 306 samples tested 3 patients (0.98%) were reported positive in both the systems i.e Matrix Gel System of Indian origin and by another CAT System of Non-Indian origin. The cost per test was comparatively cheeper in Matrix- Gel System.

Conclusion: The study concludes that both CAT Systems are equally Sensitive and Specific. Matrix Gel System and Matrix ERYGEN-AS being manufactured in India by using cells from Indian population will give equal efficacy in detecting antibodies against antigens in local population and will be cost-effective.

Hematologic and hemostatic disorders in critically ill patients: Prevalence, diagnosis and blood transfusion management at a tertiary care hospital in North India

Anupam Verma, Akshay Batra, Arvind Kumar Baronia 1 , Priti Elhence, Afzal Azim 1

Departments of Transfusion Medicine and 1 Critical Care Medicine, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow, Uttar Pradesh, India

Introduction: Severe sepsis is one of the most common disease processes that are encountered in the ICU.

A systematic evaluation of clinical and laboratory findings is necessary to ascertain the cause and prevalence of hematologic and hemostatic derangement and to determine the correct therapy. However, there is paucity of such data from our region.

Objectives: The aims of the study were: 1) to study the prevalence of various hematologic and hemostatic abnormalities in critically ill patients with severe sepsis and septic shock; 2) to identify alteration in hemostasis in critically ill patients with the help of global coagulation and viscoelastic tests and 3) to study blood transfusion management in these patients.

Materials and Methods: This prospective study was conducted, at a tertiary care hospital for two years which included critically ill adult patients who developed severe sepsis or septic shock during their ICU course. Admission diagnosis, demographic profile, co-morbidities, ICU illness severity scores like APACHE-II, SOFA were noted at admission. Laboratory data collection was done at admission and on daily basis till the end of ICU course including hematological and coagulation parameters for all patients whereas thromboelastography (TEG) was done in selected patients.

Results: Out of 394 admissions in CCM-ICU during our study period, 176 patients met inclusion criteria. Of them, 100 patients were followed; of which 82% received transfusion therapy. Overall 83% of patients were admitted with diseases related to hepatobiliary/pancreatic and respiratory systems. The mean admitting Hb was 9.1 g/dL, with 89% of patients having an admitting Hb <12 g/dL and 68% <10 g/dL. The prevalence of anemia on admission was more common than other hematological abnormalities. Patients who had higher admitting APACHE II and SOFA scores developed septic shock. Mean nadir hemoglobin (Hb) and platelet count were higher in patients with severe sepsis; whereas mean zenith TLC and INR values were higher in patients with septic shock. 89% of patients had admitting Hb less than 12g/dL. 70% had coagulopathy defined by INR more than 1.5. An average of 10 units of red cells, 20 units of plasma, and 5 units of platelets were transfused per patient during ICU stay. An average of 5 transfusion episodes (2 units of red cells, 4 units of plasma and 1 unit of platelets per episode) were given per patient. Four fifth of patients transfused prior to ICU admission had subsequent transfusions during the ICU stay. Transfusion played an important determinant role in patients' characteristics (hematologic and hemostatic) among two subgroups (severe sepsis and septic shock) during their ICU stay. Transfused patients were older, had higher weight, admitting SOFA and APACHE II scores, lower admitting Hb; and longer hospital and ICU length of stay (LOS). Patients with septic shock had higher transfusion rates. Majority of plasma transfusions were done to correct bleeding with/without coagulopathy. Red cell, platelet and plasma transfusions were more in patients who had pretransfusion Hb less than 7.0g/dL, pretransfusion platelet count between 11-50x103/΅L, and pretransfusion INR between 1.51-2.00. Red cell transfusions were more among patients' admitted due to surgical causes. Maximum platelet transfusions were done for therapeutic indication. Plasma transfusions were indicated when there were both bleeding and coagulopathy; more so in patients with septic shock. Forty one patients were categorized on the basis of clot index (CI) into 'hypocoagulable', 'normocoagulable', and 'hypercoagulable'; findings of TEG (R time, K time, angle, maximal amplitude, clot index and clot strength) were significantly different among the three categories. Majority of patients were in hypocoagulable state during their ICU stay as depicted by global coagulation tests and TEG. Platelet count correlated with findings of TEG.

Discussion: The present study validates the common occurrence of anemia in critically ill patients and also reports that lower mean Hb levels were associated with higher APACHE II and SOFA scores, longer lengths of stay, and higher mortality rates. Nadir platelet count <150 Χ 103/΅L during ICU course was found in 80% of the present cohort. This is twice than what reported by Arnold et al. and Vanderschueren et al. Coagulopathy, defined as INR >1.5, was identified in 46.0% of patients on admission (mean (SD); 2.20 (.98)). Again, Chakraverty et al. found similar prevalence of coagulopathy in their study (66.0%), A low hematocrit (<25%) was the major identifiable indication in 72% of transfusion events, far greater than 19% of transfusion events reported by Corwin et al. The indications for transfusion in the present study were acute bleeding (65.4%; 246/376) and diminished physiological reserve (32.5%; 122/376). The rate of inappropriate transfusion was 2.1% (8/376). In the present study, transfused patients had higher ICU mortality rates (94.7% vs. 5.3%) and a longer duration of hospital stay (25.3 (20.4) vs 17.1 (13.7) days; p > 0.050), higher APACHE II score, and SOFA score at admission). Also, in the present study, 82% of transfusion episodes were for 2 PRBC units. Most platelet transfusions in our study were given at values 21-50 x 103/΅L. This was different from what Rao et al. reported (50-100 x 103/΅L).

Among critically ill patients in the present study, most platelet transfusions were administered to treat, rather than to prevent, bleeding, with a wide range of transfusion trigger of 7-116 Χ 103/΅L. This was quite opposite to the findings of Arnold et al.,where most platelet transfusions were administered to prevent, rather than to treat, bleeding. This was obvious since 70% of our patients had bleeding (with coagulopathy) more than once during their ICU stay. Nearly one-third of transfusion episodes failed to mount a platelet count increase after a single transfusion in our study; less than that reported by Arnold et al. In the present study, 26% of all FFP transfusions were given in the absence of documented bleeding. This was quite less as reported (43%) in the study by Stanworth et al. Both in the present study and in the study by Holli Halset et al., significant associations were noted between the MA and platelet level on TEG.

Conclusion: The present study shows that the anemia, thrombocytopenia and coagulopathy are common hematologic and hemostatic abnormalities in critically ill patients. The prevalence of anemia on admission was more common than other hematological abnormalities. The patients with septic shock had higher incidence of hematologic and hemostatic abnormalities than the patients with severe sepsis. Majority of patients were in hypocoagulable state during their ICU stay as depicted by global coagulation tests and point-of-care test, TEG.

Further Reading

1. Levy MM, Fink MP, Marshall JC, Abraham E, Angus D, Cook D, et al. 2001 SCCM/ESICM/ACCP/ATS/SIS International Sepsis Definitions Conference. Crit Care Med 2003;31:1250-6.

2. Arnold DM, Crowther MA, Cook RJ, Sigouin C, Heddle NM, Molnar L, et al. Utilization of platelet transfusions in the intensive care unit: indications, transfusion triggers, and platelet count responses. Transfusion 2006;46:1286-91.

3. Vanderschueren S, De Weerdt A, Malbrain M, Vankersschaever D, Frans E, Wilmer A, et al. Thrombocytopenia and prognosis in intensive care. Crit Care Med 2000;28:1871-6.

4. Chakraverty R, Davidson S, Peggs K, Stross P, Garrard C, Littlewood TJ. The incidence and cause of coagulopathies in an intensive care population. Br J Haematol 1996;93:460-3.

5. Corwin HL, Parsonnet KC, Gettinger A. RBC transfusion in the ICU. Is there a reason? Chest 1995;108:767-71.

6. French CJ, Bellomo R, Finfer SR, Lipman J, Chapman M, Boyce NW. Appropriateness of red blood cell transfusion in Australasian intensive care practice. Med J Aust 2002;177:548-51.

7. Rao MP, Boralessa H, Morgan C, Soni N, Goldhill DR, Brett SJ, et al. Blood component use in critically ill patients. Anaesthesia 2002;57:530-4.

8. Stanworth SJ, Grant-Casey J, Lowe D, Laffan M, New H, Murphy MF, et al. The use of fresh-frozen plasma in England: high levels of inappropriate use in adults and children. Transfusion 2011;51:62-70.

9. Holli Halset J, Hanssen SW, Espinosa A, Klepstad P. Tromboelastography: variability and relation to conventional coagulation test in non-bleeding intensive care unit patients. BMC Anesthesiol 2015;15:28.

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2006 - Asian Journal of Transfusion Science | Published by Wolters Kluwer - Medknow
Online since 10th November, 2006