Blood Donation Screening of Transfusion-Transmissible Viral Infection Using Two Different Nucleic Acid Testing (NAT) Platforms: A Single Tertiary Care Oncology Centre Experience.

Nucleic acid testing (NAT) is used to screen transfusiontransmittable infections (TTIs) in donated blood samples and provide an additional layer of blood safety. In this study, we describe our experience in screening viral TTIs using two formats of NAT: cobas® MPX2 polymerase chain reaction- based minipool NAT (PCR MP-NAT) and Procleix Utrio Plus transcription-mediated amplificationbased individual donor-NAT (TMA ID-NAT). Data routinely collected as a part of blood bank operations were retrospectively analysed over a period of 70 months for TTIs. Blood samples were initially screened for HIV, HBV, HCV, syphillis by chemiluminescence and malaria by Rapid card test. In addition to serological testing, all samples were further screened by TMA-based ID-NAT (ProcleixUltrio Plus Assay) during Jan 2015-Dec 2016, and by PCR-based MP-NAT (Cobas® TaqScreen MPX2) during Jan 2017-Oct 2020. RESULTS: A total of 48,151 donations were processed over 70 months, of which 16,212 donations were screened by ProcleixUtrio Plus TMA ID-NAT and 31,939 donations by cobas® MPX2 PCR MP-NAT. Replacement donors and male donors outnumbered voluntary donors and female donors respectively. The overall NAT yield rate of MP-NAT was 1:2281 compared to 1:3242 with ID-NAT, during the respective time period. ID-NAT detected 5 HBV infections missed by serology, whereas MP-NAT detected 13 HBV infections and 1 HCV infection missed by serology. The proportion of donations that were both seroreactive and NAT reactive was higher with MP-NAT (59.8%) compared to ID-NAT (34.6%). Cobas® MPX2MP-NAT had higher overall NAT yield rate compared to ProcleixUtrio Plus ID-NAT and confirmed a higher proportion of seroreactive donations. Due to the ease of operation, simple algorithm, cobas® MPX2 PCR based MP-NAT can be an effective solution for blood screening in India.

Determination of the Rh/Kell phenotypes in donor as well as patients might be significant to provide phenotype-matched blood to cancer patients: A retrospective analysis from a tertiary care oncology center in North India.

BACKGROUND: Multiple reports are available from different parts of the globe indicating the incidences of alloimmunization and blood transfusion-related reactions, which emphasizes the need for phenotyping and providing antigen-matched safe blood. AIMS AND OBJECTIVES: This study aims to determine the frequency of Rh and Kell antigens and phenotype for both donors and patients to propose the importance of providing Rh Kell phenotype cross-matched packed red blood cell (RBC) units to minimize the alloimmunization and transfusion reactions. MATERIALS AND METHODS: Ten thousand blood donors and four thousand patients were investigated between October 2017 and July 2019. Each donor unit was tested for blood grouping, antibody screening, and Rh Kell antigen Phenotyping, and the blood unit was issued after the patient's blood grouping, antibody screening by 3 cell panels, and Rh Kell antigen phenotyping followed by cross-matching with an Rh Kell-matched phenotype RBC unit. RESULTS: Nine thousand four hundred and fifty-two donors were D positive (94.5%) while 548 tested D negative (5.5%). Overall Rh and K antigens frequencies in donors were: "e" (98%) >"D" (94.5%) >"C" (86.6%) > "c" (57.5%) >"E" (18.8%) >K (0.98%). Among patients, 3762 tested D positive (94.05%), and 238 tested D negative (5.95%). Overall Rh and K antigens frequencies in patients were: "e" (98.5%) >"D" (94.05%) >"C" (90.2%) >"c" (51%) >"E" (18.2%) >K (1.8%). CONCLUSION: Our study has given us more clarity on the prevalence of major Rh and K antigens in our donor as well as patient populations, highlighting the similarities as well as differences. This variance holds a great significance, since such donor units when transfused into patients may lead to alloimmunization and adverse transfusion reactions. Hence, the determination of Rh and Kell phenotypes and providing phenotype-matched blood will help prevent such events.

Benefits of Pre-harvest Peripheral Blood CD34 Counts Guided Single Dose Therapy with PLERIXAFOR in Autologous Hematopoietic Stem Cell Transplantation: A Retrospective Study at a Tertiary Care Institute in India.

Peripheral blood is a convenient source of stem cells for hematopoietic stem cell transplantation. However, in autologous transplants, the harvest failure rates are high because of inadequate mobilization using G-CSF alone. Plerixafor is a potent mobilizer when used with G-CSF. However, its routine use is limited by high cost. This is a retrospective study done at a tertiary care oncology centre in India. All the harvest records were analyzed between Jan 2015 and Nov 2017. May 2016 onwards pre-harvest peripheral blood CD34 count was done in all cases of autologous transplants on day 4 of G-CSF therapy and they were given a single dose of Plerixafor if counts were < 20 cell per cumm. The results were compared amongst various groups. A total of 321 cases were analyzed. 172/321 were allogenic transplant cases of which 5% (n = 7) failed to achieve a target live stem cell dose of > 2 million per kg of the recipient. The overall failure rate in autologous group (n = 149) was 27% (n = 41) (p ≤ 0.001 auto vs. allo). The failure rate was higher (36%, n = 28/77) when no intervention with Plerixafor was done. The overall failure rate in the group treated with pre-harvest 34 count based single dose therapy of Plerixafor was 18% (n = 13/72, p = 0.01). However, within this intervention group, the patients who had pre-harvest peripheral blood CD34 above the desired cutoff had a higher failure rate of 21% (p = 0.13). Pre-harvest CD34 count based intervention with Plerixafor help optimizing the cost.

Robotic hysterectomy in Trendelenburg position in a severely anaemic JKa alloimmunised patient with impending high-output cardiac failure: An anaesthetic challenge.

Kidd blood group alloimmunisation, though extremely rare, may produce considerable morbidity, and even mortality. Severe anaemia and impending high-output cardiac failure requiring blood transfusion should be weighed against the risk of severe transfusion reactions even with fully cross-matched blood. Kidd antibodies are a common cause of delayed haemolytic transfusion reaction (DHTR) since they have a tendency remain undetectable in plasma. A low -grade DHTR (second hit) was grossly amplified by a second DHTR (third hit) superimposed on it in our patient leading to severe haemolysis with serum bilirubin reaching 68 mg%. Indirect antiglobulin test (indirect Coombs reaction) should ideally be performed in all patients (scheduled for major surgery requiring blood transfusion) who have experienced a previous pregnancy or blood transfusion. Non-invasive continuous haemoglobin monitoring and non-invasive cardiac output monitoring can prove invaluable tools in management.

Comparison of antibody titers using conventional tube technique versus column agglutination technique in ABO blood group incompatible renal transplant.

INTRODUCTION: Measurement of alloantibody titer to a red cell antigen (ABO titers) is an integral part of management of ABO incompatible kidney transplants (ABOiKT). MATERIAL AND METHODS: There are different methods of titer estimation. Alloantibody detection by tube titration and Gel agglutination columns are accepted methodologies. It is essential to find the difference in titers between the two methods so as to set the 'cut-off' titer accordingly, depending upon the method used. RESULTS: We did a prospective observational study to compare and correlate the ABO titers using these two different techniques - conventional tube technique (CTT) and the newer column agglutination technique (CAT). A total of 67 samples were processed in parallel for anti-A/B antibodies by both tube dilution and column agglutination methods. The mean titer by conventional tube method was 38.5 + 96.6 and by the column agglutination test was 96.4 + 225. The samples correlated well with Spearman rho correlation coefficient of 0.94 (P = 0.01). CONCLUSION: The column agglutination method for anti A/B titer estimation in an ABO incompatible kidney transplant is more sensitive, with the column agglutination results being approximately two and half fold higher (one more dilution) than that of tube method.

The effect of repeated freezing and thawing on levels of vitamin K-dependent coagulation factors and fibrinogen in fresh frozen plasma.

BACKGROUND: Fresh frozen plasma (FFP) is considered adequate for transfusion immediately after thawing or for up to 24 hours if kept at 1-6°C, and is currently used very often to replace deficient clotting factors. If factor levels in refrozen FFP are within normal limits, then this component can possibly be transfused, thus avoiding wastage of FFP. AIM: To study the fate of vitamin K-dependent coagulation factors (F II, F VII, F IX, F X) and fibrinogen activity levels in repeatedly (twice) frozen and thawed FFP. MATERIALS AND METHODS: Two hundred FFP units comprising 50 units of each major blood group (A, B, AB, and O) were thawed at 37°C and 10-20 mL of FFP transferred to transfer bags with the help of a sterile connecting device (SCD). The FFP samples were taken into tubes (first sampling), and then the transfer bags were kept for 24 hours at 4°C. After 24 hours, repeat samples were taken in tubes from the transfer bag (second sampling), and then the bags were re-stored at < -18°C. One week later, the above procedure was repeated. Activity of coagulation factors and fibrinogen levels were measured by the automated coagulation analyzer. RESULTS: The levels of F II, F VII, F IX, F X, and fibrinogen of all the 200 FFP units, at all four time points, were above the lower normal value, but well within the normal range. CONCLUSION: The levels of F II, F VII, F IX, F X, and fibrinogen remain stable and adequate for transfusion in twice-thawed-and-refrozen FFP. This component can be safely used for transfusion as a source of vitamin K-dependent clotting factors and fibrinogen.

Adverse events associated with apheresis procedures: Incidence and relative frequency.

INTRODUCTION: Apheresis procedures [Plateletpheresis, Plasmapheresis/ Therapeutic Plasma Exchange (TPE), & Peripheral Blood Stem Cell Collection (PBSC)] are usually well tolerated. Occasionally, Adverse Events (AEs) of variable severity may occur during or after the procedure. AEs that occur in Donors/Patients are divided into local reactions and systemic reactions. MATERIALS AND METHODS: A total of 3,367 apheresis procedures were performed, out of which 3,120 were plateletpheresis procedures, and out of which 1,401 were on Baxter CS 3000 & 1,719 were on Haemonetics MCS+ cell separators. Rest of 247 TPE & PBSC procedures were done on Haemonetics MCS+ cell separators. RESULTS: 90 AEs were reported in relation to the 3,367 procedures. Out of 90 AEs, 85 AEs (94%) were associated with plateletpheresis (n = 3,120) and 05 AEs (06%) with TPE & PBSC (n = 247). The rate of vascular injury (VI), Citrate reaction (CR), and Presyncopal/Syncopal (PS/S) in plateletpheresis was 1.6% (52/3,120), 0.96% (30/3,120), and 0.096% (03/3,120), respectively. The rate of CR in TPE and PBSC was 1.23% (02/162) and 2.3% (02/85), respectively. The rate of PS/S in PBSC was 1.17% (01/85). AEs for Plateletpheresis, TPE & PBSC were 2.7% (85/3,120), 1.23% (02/162), and 3.5% (03/85), respectively. VI, CR, and PS/S were mostly of mild intensity. Both cell separators were equally safe, when AEs associated with plateletpheresis were compared with each other; 2.8% on CS 3000 & 2.6% on MCS+. CONCLUSION: Apheresis procedures performed on cell separators are safe, with a low incidence of significant AEs. No significant difference was noted in AEs among the two cell separators studied.

Effect of gamma irradiation with 30 gy on the coagulation system in leukoreduced fresh frozen plasma.

BACKGROUND: Fresh Frozen Plasma (FFP) is a blood component prepared from whole blood or from apheresis donation. Donor leukocytes including lymphocytes are present in FFP in significant numbers inspite of freezing, responsible for Transfusion Associated Graft versus Host Disease (TA-GvHD). STUDY DESIGN AND METHODS: 75 units of FFP prepared at our centre were analysed. After thawing of FFP a small aliquot was made under sterile conditions and another after irradiating the product. In both the parts, variables of haemostasis were measured in parallel, using automated coagulation analyser. Activated partial thromboplastin time (APTT), prothrombin time (PT), International Normalized Ratio (INR), Thrombin time (TT), coagulation factors FI, FII, FV, FVIII, FIX, FX, FXI, FXII, vWF Ag, inhibitors of coagulation (protein C & S) and d-dimer were measured. RESULTS: Gamma irradiation of FFP with 30 Gy resulted in weak activation of coagulation system which was evident in the form of shortening of PT, APTT and TT. The activity of coagulation factors FIX, FX, FXI, and FXII were significantly raised after irradiation. No reduction in the activity of inhibitors of coagulation (protein C & S) or increase in d-dimers was observed following irradiation of FFP. CONCLUSION: Gamma irradiation of FFP with 30 Gy resulted in a significant but very weak alteration of coagulation system in FFP.

Changing trends of transfusion transmitted viral infections among blood donors in the last decade-a 10-year study in a large tertiary care blood bank (2000-2009.

BACKGROUND: The main blood borne viruses' viz. hepatitis B virus (HBV), human immunodeficiency virus (HIV), and hepatitis C virus (HCV), are a major public health issue, and represent significant causes of morbidity and mortality associated with transfusion. This study analysed the trends of blood borne infections among blood donors in a large blood bank in the last 10 years. METHOD: Viral screening results of 80,500 voluntary and replacement donations from 2000 to 2009 were analysed. All donations were screened for HBV, HCV, and HIV. The seroprevalence rate of HBV, HCV, and HIV infections and 95% confidence interval were calculated. RESULTS: The seroprevalence rate of HBV, HIV, and HCV, decreased during the last 10-years study from 2000 to 2009. There is significant and impressive decrease in HBV seroprevalence rate from 2.39% in 2000 to 1.28% in 2009. The seroprevalence rate of HIV appeared to have decreased with a very significant value from 1.32% to 0.30% in 2009. Hepatitis C virus seroprevalence rate showed a slight decline in blood donations from 0.48% in 2001 to 0.22% in 2009. CONCLUSION: There is a general reduction in the seroprevalence rate of viral infections. This is probably because of discouragement of professional or paid donations; better awareness, better prophylactic measures, and availability of vaccines have played a major role.