Asian Journal of Transfusion Science
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   Table of Contents - Current issue
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July-December 2022
Volume 16 | Issue 2
Page Nos. 159-288

Online since Saturday, November 12, 2022

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REVIEW ARTICLE  

Application of flow cytometry in transfusion medicine: The Sanjay Gandhi Post Graduate Institute of Medical Sciences, India experience Highly accessed article p. 159
Rajendra Chaudhary, Sudipta Sekhar Das
DOI:10.4103/ajts.ajts_61_22  
The application of flow cytometry (FC) is diverse and this powerful tool in used in multiple disciplines such as molecular biology, immunology, cancer biology, virology, and infectious disease screening. FC analyzes a single cell or a particle very rapidly as they flow past single or multiple lasers while suspended in buffered solution. FC has a great impact in the field of transfusion medicine (TM) due to its ability to analyze individual cell population and cell epitopes by sensitive, reproducible, and objective methodologies. The main uses of FC in TM are detection of fetomaternal hemorrhage, diagnosis of paroxysmal nocturnal hemoglobinuria, quantification of D antigen, detection of platelet antibody, quality control of blood components, for example, residual leukocyte counts and evaluation of CD34-positive hematopoietic progenitor cells in stem cell grafts. In recent years, FC has been implemented as an alternative method for the detection and characterization of red cell autoantibodies in autoimmune hemolytic anemia. Many workers considered FC as a very good complement when aberrant expression of various erythrocyte antigens needs to be elucidated. It has been extensively used in the resolution of ABO discrepancies and chimerism study. FC has also been used successfully in various platelet immunological studies. In the recent past, FC has been used in several studies to assess the platelet storage lesions and elucidate granulocyte/monocyte integrity and immunology. FC analysis of CD34+ stem cells is now the method of choice to determine the dosage of the collected progenitor cells. The technique is vastly used to evaluate residual leukocytes in leukodepleted blood components. We conclude that flow cytometers are becoming smaller, cheaper, and more user-friendly and are available in many routine laboratories. FC represents a highly innovative technique for many common diagnostic and scientific fields in TM. Finally, it is the tool of choice to develop and optimize new cellular and immunotherapeutic trials.
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ORIGINAL ARTICLES Top

Distribution and frequency of principal Rh blood group antigens (D, C, c, E, and e) and their phenotypes in the blood donors attending blood bank in a tertiary care hospital in Barpeta district of Assam p. 167
Dipankar Baruah, Gitali Devi, Jabin Musfique, Abhijit Bharali, Umesh Chandra Dutta
DOI:10.4103/ajts.AJTS_64_20  
BACKGROUND: The ABO and Rhesus grouping system antigens have been found to have the highest immunogenicity and propensity to produce alloantibodies that cause most of the transfusion reactions. The Rhesus antigens that produce most of the immunogenic transfusion reactions are D, C, c, E, and e. Knowledge of the distribution of these Rh antigens in a population helps to render compatible blood in alloimmunized patients. AIM: The aim was to study the distribution and frequency of principal Rh blood group antigens (D, C, c, E, and e) and their phenotypes in the blood donors attending blood bank in a tertiary care hospital in Barpeta district of Assam. MATERIALS AND METHODS: The study was conducted in 315 voluntary blood donors in the blood bank of a tertiary care center. Rh-D typing was done by conventional tube method. Specific monoclonal antisera, i.e., anti-C, anti-c, anti-E, and anti-e, were used and tests were performed by conventional tube method for detection of the presence of rest of the major Rh antigens. RESULTS: The samples were analyzed for the five major Rhesus antigens. “D” antigen was found to be the most common antigen (99.05%), followed by e (97.14%), C (92.38%), c (51.43%), and E (20.95%). In order of descending frequency, the most common phenotypes were DCCee – 45.71%, DCcee – 30.48%, DCcEe – 11.43%, DccEe – 4.76%, DCcEE – 1.90%, DCCEe – 1.90%, Dccee – 1.90%, DCCEE – 0.95%, and dccee – 0.95%. CONCLUSION: D antigen is the most common antigen in our study population, whereas “e” antigen is the most common in most of the studies done from other parts of India. Data on frequencies of major Rh antigens in the local donor population will help in transfusing alloimmunized patients with corresponding antibody-negative blood ensuring blood safety.
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Factor assay in victims of snake bite: Experience from a tertiary care institute in South India p. 175
Aboobacker Mohamed Rafi, Susheela Jacob Innah
DOI:10.4103/ajts.AJTS_104_18  
INTRODUCTION: Snake bites tend to cause a high mortality in those who develop coagulopathy. However, there is very limited literature on clotting factor assays in these patients, especially in the presence of clinical bleeding. The aim was to assess the coagulation profile and individual coagulation factors in patients with hematotoxic snake bites. MATERIALS AND METHODS: This was a prospective observational study of clotting factor levels in victims of snake bites with hematotoxicity admitted to a single hospital in south India for 12 months. In 43 individuals who fulfilled the criteria, we measured platelet count, prothrombin time (PT), international normalized ratio, activated partial thromboplastin time (aPTT), fibrinogen levels, coagulation factors V, VII, VIII, IX, and X, and the qualitative factor XIII assay. RESULTS: Forty-three patients fulfilled the criteria and their samples were studied. There were 36 Russell's viper (Daboia russelli), 4 Hump-nosed pit viper (Hypnale hypnale), and 3 unknown snake bite victims samples, in which factor assays were done. All the Russell viper bite victims without a recordable clotting screen had deficiency of Factor V (0.5%–49.62%, Mean – 20.27%), Factor X (0.08%–92.3%, Mean – 70.73%), and qualitative factor XIII. Pit viper patients showed normal levels of Factor I, V, VII, VIII, IX, X, and XIII despite prolonged PT and aPTT. CONCLUSIONS: Early detection and treatment of envenomation remains the cornerstone of managing snake venom-induced consumptive coagulopathy. Anti-snake venom plays a major role in the reversal of coagulopathy. Blood and blood products would be useful when coagulopathy does not revert by ASV alone. Evidence-based transfusion can be implemented and cryoprecipitate may be used as many of the patients had factor XIII and fibrinogen deficiency as part of venom-induced coagulopathy. To improve patient management and thereby the outcome of patients CMEs and training programs for the treating physicians also has to be implemented so that guidelines are formulated and followed.
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Prozone phenomenon in pretransplant testing: An interesting conundrum involving solid-phase and cell-based assays p. 180
Parvind Singh, Aseem Kumar Tiwari, Vikash Chandra Mishra, Trupti Vimlakar Deshpande, Pranav Dorwal, Amit Kr Bhardwaj, Sneha Kumari, Vimarsh Raina
DOI:10.4103/ajts.ajts_145_20  
BACKGROUND: Human leukocyte antigen (HLA) is a major determinant in deciding upon solid organ histocompatibility. Donor-specific anti-HLA antibodies (Donor-specific anti-HLA antibodies (DSAs)) are always a contraindication for solid organ transplantation, and identification of DSA becomes very crucial before transplantation to provide long-term graft survival. For identification of DSA, usually, either cell-based or HLA bead-based assay is being used in laboratories. However, both cell-based and bead-based assays have certain limitations. One such common limitation is “prozone effect,” which can give false-negative results. Here, we would like to present a small pilot study to analyze the effect of the prozone phenomenon in the cell-based and HLA bead-based assays and its utility in histocompatibility testing. MATERIALS AND METHODS: In a series of four experiments, cell-based assay, flow cytometric cross-match (FCXM), and HLA bead-based flow cytometric panel reactive antibodies (PRAs) were performed. Single-antigen bead (SAB) testing was conducted as a first experiment on four known positives samples for anti-HLA antibody-antibodies. In the second experiment, these four samples were pooled together (called pooled sera in the text) and tested for FCXM and PRA. In the third experiment, known commercially available positive control sera were mixed with pooled positive sera (positive control sera + pooled sera) to prepare, what we have called “positive concoction” in the text. In the fourth experiment, the positive concoction was diluted serially (1:2, 1:4, 1:8, and 1:16) and FCXM and PRA were performed again to analyze and compare the prozone effect. RESULTS: Pooled sera did not have the expected median fluorescence intensity (MFI) values in FCXM assay, whereas the PRA was showing >90% positivity. In positive concoction, the MFI of FCXM assay was observed to be declining; however, PRA values remained almost constant. Dilutions of the pooled sera showed that MFI values of FCXM assays were increased suddenly after dilution. The highest MFI values were observed in 1:4 dilution of the sera, and then, it declined gradually, but the PRA values remained almost constant even after serial dilutions. CONCLUSION: In our experimental findings, it was clear that cell-based assay (FCXM) was more severely affected by the prozone, whereas solid-phase (flow PRA) assay remained resistant to prozone.
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Assessment of iron status in regular blood donors in a tertiary care hospital in Southern India p. 186
Joy Anju, Basavarajegowda Abhishekh, Basu Debdatta, Zachariah Bobby, Murali Sharan
DOI:10.4103/ajts.ajts_119_21  
BACKGROUND: Regular blood donation depletes iron stores. The assertion is that the vulnerable donor population requires a predictive standard operative procedure for early detection of iron store depletion, preventing them from developing iron-deficiency anemia. AIM: This study aims to study the potential effects of blood donation in the regular donor group using hematological and biochemical estimation of iron status parameters. STUDY SETTINGS AND DESIGN: This was a prospective cross-sectional study on regular blood donors, defined as those who have donated at least 3 times, the last donation being within the last 12 months and continues to donate at least once a year, at a tertiary care teaching hospital in Southern India. MATERIALS AND METHODS: The complete blood count (CBC) was performed on the Sysmex coulter, and the red cell indices were calculated. The ferritin and the soluble transferrin receptor (sTfR) assays were performed using Enzyme Immunoassays. STATISTICAL ANALYSIS USED: The comparison of CBC, serum ferritin, and sTfR assay with donation frequency and time since the last donation was carried out using an independent student's t-test for two groups. The statistical analysis was performed using SPSS for Windows version 20. RESULTS: A total of 323 regular blood donors (6 were females) were included in the study of which they were categorized into three, 211 donors with less than or equal to 10 donations, 84 those who had donated between 11 and 20 times and 28 who had donated more than 20 times. The red cell indices were reduced and different in the groups but not statistically significant except for mean corpuscular volume. About 15% of the study population had a transferrin level of <15 ng/ml. The Ferritin levels showed a statistically significant negative correlation with the number of donations, the correlation coefficient being –0.27. Logarithmic ratios of sTfR/ferritin also correlated with a coefficient of 0.156 with the number of donations and were statistically significant. CONCLUSION: Our study found that regular blood donors had low iron stores, as shown by ferritin levels and other iron indicators. Using the current guidelines (hemoglobin >12.5 g/dL) for donation, or the red cell indices alone do not reflect the donor's actual iron status.
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An insight to the internal quality control of blood components separated using the latest whole blood collection and processing systems: Experience from a tertiary care hospital blood transfusion service in Eastern India p. 194
Sudipta Sekhar Das, Rathindra Nath Biswas, Tirtha Pratim Sardar, Mahammad Safi
DOI:10.4103/ajts.ajts_52_21  
BACKGROUND: With blood component therapy becoming the standard of care in transfusion medicine globally, the quality control (QC) of these components has become a routine and mandatory program in all blood centers. Extensive utilization of blood components has been observed in our multidisciplinary tertiary care hospital. We use quadruple bag systems and automated component extraction facilities for collection and processing of whole blood (WB). In this study, we analyzed our data relating to QC of all blood components which we prepare and issue for transfusion. MATERIALS AND METHODS: The retrospective 5-year study comprised 47,430 WB collections which were separated into blood components using quadruple bags and automated component extraction machine. A total of 90 units of WB were processed into blood components for the machine calibration and validation. Routine use of the system was started once the calibration and validation results were acceptable. At least 1% of each component prepared was subjected to QC as per departmental standard operating procedures. Statistical analysis was done using the SPSS statistical package. RESULTS: The mean volume, hematocrit (Hct), platelet (PLT), and white blood cell (WBC) in 350 and 450 mL WB units were 394.63 mL, 39.43%, 0.93 × 1011, and 3.12 × 109 and 507.75 mL, 40.72%, 1.13 × 1011, and 3.45 × 109, respectively, with mean recovery of PLT and WBC in buffy coat being 95.54% and 68.63% and 97.87% and 74.51%, respectively. As high as 89.91% RBC recovery was noted in the packed red blood cell units which were subjected to QC. QC of random donor platelets was performed in 979 (2.36%) units with acceptable results. The mean fibrinogen and FVIII values were estimated to be 469.17 mg and 217.34 IU (1.07 IU/mL) and 600.21 mg and 273.39 IU (1.11 IU/mL) in fresh frozen plasma units prepared from 350 and 450 mL WB, respectively. A total of 578 (1.62%) units of cryoprecipitate were investigated for QC with favorable results. CONCLUSION: We conclude that QC data generated in this study will provide invaluable information about the performance of the latest blood collection systems. QC of all blood components under study complied with both national and international standards. We opine that all blood centers should establish a complete QC program and adhere to departmental protocols and manufacturer's instructions for its execution and effective outcome.
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Assessment of noncompliance in self-disclosure of deferrable risk behaviors among blood donors p. 201
Bala Vignesh Venkatachalam, Ravindra Prasad Thokala, Ashwin Anandan, Krishnamoorthy Radhakrishnan
DOI:10.4103/ajts.AJTS_119_20  
BACKGROUND: Blood transfusion services work to ensure universal accessibility of safe and effective blood products for transfusion to recipients. Failure of blood donors to disclose complete truthful information before blood donation is termed as noncompliance. Noncompliance in disclosing high-risk behaviors could compromise blood safety. This study aimed to assess the prevalence rate of noncompliance and assess the predictive factors and reasons for noncompliance. MATERIALS AND METHODS: Blood donors were asked to fill a postdonation anonymous questionnaire after obtaining consent and the responses were tabulated and analyzed. Prevalence of noncompliance for both high-risk and nonhigh-risk behaviors are evaluated. Variables associated with noncompliance are analyzed by univariate analysis and logistic regression. RESULTS: Total number of participants was 3001, 2850 participants gave valid responses and included in the study. There were 94 (3.30%) responses revealing noncompliance for nonhigh-risk behavior and 30 (1.05%) responses revealing noncompliance for high-risk behavior. The predictor variables for noncompliance in reporting high-risk behavior were education and adultery. The predictor variables for noncompliance in nonhigh-risk behavior reporting were presence of comorbidity and adultery. CONCLUSION: Noncompliance in disclosure of high-risk behavior compromises blood safety. Blood donors must be ensured sufficient privacy while filling predonation questionnaire and while eliciting history any deferrable behaviors during blood donor medical examination. Privacy and confidence of the donors must be ensured either to share any postdonation information directly or anonymously to facilitate confidential unit exclusion.
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A preliminary experience of plasma exchange in liver failure p. 209
Himanshu Dandu, Vivek Kumar, Amit Goel, Dheeraj Khetan, Tulika Chandra, Vipin Raj Bharti
DOI:10.4103/ajts.ajts_115_21  
INTRODUCTION: Plasma exchange (PLEX) is one of the experimental modalities of treatment for liver failure. We report our experience of PLEX in patients with acute-(ALF) or acute-on-chronic (ACLF) liver failure. METHODS: Hemodynamically stable adult patients with ALF or ACLF, encephalopathy, model for end-stage liver disease (MELD) score ≥ 15, and clinical worsening/no improvement after 72-h of inpatient care were included. PLEX cycles repeated every 48 h, each of 2.5–4.0 h duration with 1–1.5 times of estimated plasma volume, were given. PLEX cycle was repeated till either of the end-points were achieved (i) MELD < 20 for 48 h or reaches below the baseline, whichever is lower, (ii) completed three PLEX cycles, (iii) hemodynamic instability, (iv) or outcome achieved. Outcome of interest was categorized as favorable (discharged in stable condition) or unfavorable (death or discharge in moribund condition). Data are expressed as median (interquartile range). RESULTS: Sixteen patients (age 35 [27–48] years; male 8; ALF 5, ACLF 11; MELD 33 [27–37]; CLIF-SOFA 10 [8.5–12]) were included. Participants received 2 (1-3) cycles of PLEX during 13 (11–25) days of hospitalization. Overall, serum bilirubin, INR, creatinine, MELD, and CLIF-SOFA scores were significantly improved after PLEX. Five patients (5/16, 31%) had complete resolution of HE. Eight patients (50%) had a favorable outcome. Those with favorable outcome had significant improvement in serum bilirubin, INR, and CLIF-SOFA scores as compared to those with unfavorable outcome. CONCLUSION: PLEX may be effective in patients with ALF or ACLF. More data are needed to establish its role in the management of liver failure.
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Plateletpheresis donor deferral pattern: A retrospective 4-year data analysis at tertiary care center in India p. 214
Brijesh Kumar Yadav, Harsha Shrivastava, Rahul Katharia, Rajendra K Chaudhary
DOI:10.4103/ajts.ajts_96_22  
BACKGROUND: Increasing demand of single donor platelet requires blood banks to expand the donor pool. A reassessment of donor deferral criteria for plateletpheresis is required to ensure that this increasing demand is met without compromising on product quality and donor safety. AIMS: (1) To list the various causes of SDP donor deferral. (2) To discuss various approaches to minimize it. MATERIALS AND METHODS: Data of plateletpheresis donor deferral were collected from records retrospectively over a period of 4 years from January 2017 to December 2020. STATISTICAL ANALYSIS: All statistical tests were performed using IBM SPSS software for Windows version 20. Categorical variables were presented as proportions, while continuous variables were presented as mean with standard deviation, mean calculated P < 0.05 was considered statistically significant. RESULTS: Out of the 7478 donors screened for plateletpheresis procedure, 3232 (43.2%) were deferred among which 3089 (42.5%) were male and 142 (63.1%) were female donors. Majority (96.5%) of deferral were temporary. These included low platelet count (47.4%) followed by poor venous access (22.4%) and low hemoglobin (Hb) (7.2%). Among the donors deferred for low Hb, 24.7% (58 out of 234) had Hb between 12 and 12.4 g%. Similarly, among donor deferred for low platelet count, 12% (184 out of 1532) had platelet count between 140 and 149 × 103/μl. CONCLUSION: There is potential for increasing the number of eligible plateletpheresis donors if the present donor selection criteria were relaxed to a minimum Hb of 12 g/dl and minimum platelet count of 140 × 103/μl.
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Major and minor subgroup profile of blood in patients receiving multiple transfusions and donors p. 219
Mehmet Fatih Orhan, Merve Pilavci Adigül, Mustafa Altindiş, Mehmet Köroğlu
DOI:10.4103/ajts.ajts_17_21  
OBJECTIVE: It was aimed to profile the blood subgroups of our region and to reveal the prevalence of auto/alloimmune sensitization in patients who had to undergo multiple erythrocyte transfusions and to establish the sensitization profile by screening major and minor subgroups. MATERIALS AND METHODS: In this study, the distribution of ABO and Rh system major subgroups was studied in 100 donor blood. As the patient group, 50 patients who received three or more red blood cell transfusions were included. In addition to this group, Kell, Lewis, Duffy blood group systems were studied. RESULTS: According to the ABO system, 35% of the donors were in O, 33% in A, 17% in AB, and 15% in B. According to the Rh system, 75% is Dvi positive. Rh system is 99% e positive and 33% E positive in major subgroups and Kell1 positivity is 8%. In the patient group, 22% D(-) was determined compared to Rh blood group. Among the major subgroups of Rh, C was 68%, E was 14%, c was 76%, and e positivity was found to be 100%. The Kell1 negativity rate is 96%. The highest negativity was found in 86% Lea antigen in Lewis system, in 36% S antigen in MNS system, 34% Fyb antigen in Duffy system, and 24% Jka antigen in Kidd system. When inappropriate blood is given for any antigen, a double population is formed. The double negativities we detected in our study occurred as follows according to blood group systems: E 18%, C 12%, c 8%, Cw 2%, Kell 1 2%, M 8%, N 4%, S 18%, s 6%, Fya 8%, Jka 6%, Jkb 22%. Indirect Agglutination Test (IAT) was negative in all patients. CONCLUSION: IAT negativity in all patient groups suggests that we do not develop alloimmunization, but the high rates of double population suggest a high risk of alloimmunization.
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Ramadan favors first blood donation, but not frequent donation: Results of 10,145 blood donors from Algeria p. 224
Mounir Ould Setti, Djamal-Eddine Damerdji, Abdelkader Nebab, Ari Voutilainen
DOI:10.4103/ajts.ajts_166_20  
CONTEXT: Frequent blood donors contribute to an important share of blood donations in many countries. In Algeria, frequent donation and its determinants, notably the place of the month of Ramadan, which plays an important role in blood donation in Muslim countries, have not been studied. METHODS: This was a retrospective cohort study of n = 10145 Algerians who donated blood to the blood transfusion post (BTP) of Boufarik between January 2, 2008, and December 31, 2019. Donors were assessed at each donation for general clinical information, demographic information, and dates and times of donation. Donor return, defined as two or more donations to Boufarik BTP, and frequent donation, defined as three or more blood donations to Boufarik BTP, were the outcomes of interest and were analyzed using groups comparison and logistic and Cox regression analyses. RESULTS: 2.2% of donors were frequent donors and donated 9.6% of all donated blood. The volume of donated blood during Ramadan was twice the monthly volume during the rest of the year, but donation in Ramadan was associated with lower odds of return (odds ratio [OR]: 0.54, 95% confidence interval [CI]: 0.40–0.71) and frequent donation (OR: 0.41, 95% CI: 0.24–0.73). Women were underrepresented (10.9%), but they were more likely to be frequent donors (male vs. female OR: 0.55, 95% CI: 0.31–0.96; hazard ratio: 0.64, 95% CI: 0.41–0.98). CONCLUSIONS: Reducing the gender gap and promoting return could significantly improve the volume of blood donations in Algeria.
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Internal quality control for HIV testing of blood donors - Dried tube specimen as a cost-effective alternative p. 231
Ashish Dhoot, Joy J Mammen, Nitty S Mathews, Rajesh Kannangai, Dolly Daniel, S Prasannakumar
DOI:10.4103/ajts.ajts_75_21  
BACKGROUND: An important aspect of ensuring blood safety is the performance of mandatory serological testing for transfusion transmissible infections. The practice of internal quality control (IQC) in blood banks in India is nonuniform, especially the use of third-party materials. Cited reasons are cost, lack of access to control materials, and need for deep-freezers for storage, if prepared in-house. OBJECTIVE: Validation of dried tube specimen (DTS) from HIV-positive plasma as a low-cost, stable material for use as IQC material in blood banks. METHODS: Fresh-frozen plasma (FFP) prepared from four HIV-positive blood-donors were pooled. Equal numbers of seronegative FFPs were pooled. Twenty microlitre aliquots of plasma were made in micro-centrifuge tubes and air-dried overnight at room-temperature. These were stored in 2–8°C refrigerators and tested once weekly for 6 months on multiple platforms with different detection principles: Rapid tests, second-generation enzyme-linked immunosorbent assay (ELISA), fourth-generation ELISA, and fourth-generation Chemiluminescence immunoassay. The protocol was sustained over the next 6 months with decreased testing frequency to study the extended stability of DTS. RESULTS: A total of 139 positive-DTS and 139 negative-DTS were tested with 100% samples showing consistent results on all platforms over 1 year. There was mild deterioration in reaction strengths, which did not interfere in result interpretations. CONCLUSION: Plasma in form of DTS maintained stability when stored at 2–8°C for 1 year. This provides evidence that DTS can be a modality for the production of cost-effective, stable, in-house control material for resource-restricted countries.
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Platelet additive solution suspended apheresis platelets in a tertiary care hospital: A step toward universal single donor platelets p. 238
Mohit Chowdhry, Soma Agrawal, Bindu Prakash, Uday Kumar Thakur, Manoj Mishra
DOI:10.4103/ajts.ajts_145_21  
BACKGROUND: Transfusion of ABO-compatible single donor platelets (SDP) is preferable for better outcomes over group switchover SDP. The use of SDP containing ABO-incompatible plasma is associated with a risk of allergic and acute hemolytic transfusion reactions. Moreover, high titer O group donors SDP impose a further threat to patient safety. Platelet additive solution (PAS) is used worldwide for the storage of platelets which reduces plasma volume available in SDP. SSP + (Macopharma) is one such PAS which can provide improved availability, logistical management, decrease wastage, and improvement in patient safety. The aim of this study was to assess the feasibility of using PAS to obtain low titer SDP units which can be utilized across a larger patient population and to study quality control parameters of these units. MATERIALS AND METHODS: The study was performed in the department of Transfusion Medicine from June 2017 to January 2018 after clearance from the Institutional Review Board. The study design comprised two cohorts (A and B). In cohort A, the temporal trend of in-vitro changes in the quality parameters was tested and analyzed for PAS modified and unmodified products on days 1, 5 and 7. In cohort B, the original plasma from the SDP donors of all blood group donors except the AB group was tested for antibody titers before (prepreparation) and after modification (postpreparation) by PAS. RESULTS: In cohort A, in the control group, there was a significant change in the mean platelet volume, potassium, and bicarbonate levels from day 1 to day 7, whereas no significant change in the biochemical parameters was noted in the study group where PAS was used. In cohort B, on comparing the anti-A and anti-B, before and after modification of SDP with PAS, there was a significant reduction in the median titers across all the groups studied. CONCLUSION: PAS added SDP is an efficient strategy to reduce the ABO-antibody levels significantly. PAS added SDP also helps in the better inventory management of available groups.
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Reference interval of platelet counts and other platelet indices in apparently healthy blood donors in North India according to Clinical and Laboratory Standards Institute guidelines: Need to redefine the platelet count cutoffs for repeat plateletpheresis donation? p. 245
Swati Pabbi, Aseem Kumar Tiwari, Geet Aggarwal, Govind Sharma, Arghyadeep Marik, Aanchal Sunil Luthra, Anand Prakash Upadhyay, Manish Kumar Singh
DOI:10.4103/ajts.AJTS_121_20  
BACKGROUND: In clinical practice, laboratory results are of great importance for the diagnosis and treatment. Reference intervals of different parameters aid health-care professionals in the interpretation of results. There are very few studies on reference intervals from India. This prospective study was conducted to determine the reference intervals for platelet count (PLT) and PLT indices; mean PLT volume (MPV), PLT distribution width (PDW), and PLT large cell ratio (P-LCR). These values can be obtained as a part of a routine complete blood count (CBC) and have diagnostic and prognostic significance in certain diseases. PLT count is an important criterion for the selection of donors for repeat plateletpheresis donation. MATERIALS AND METHODS: Sixteen hundred and thirty-four first-time healthy volunteer plateletpheresis donors were enrolled for the study. CBC was done, values of PLT, MPV, PDW, and P-LCR were noted, and the results were analyzed. The 95% of the reference distribution was estimated using the 2.5th and 97.5th percentiles following Clinical and Laboratory Standards Institute guidelines. Adverse donor reactions, if any and quality parameters of single donor PLTs (SDP) were also studied. RESULTS: Reference range values of PLT, MPV, PDW, and P-LCR were 137,825–355,175/μl, 8.1–13.9/fl, 9.1–22.5/fl, and 11.7%–52.9%, respectively, and compared well with other published studies from India. It was observed that reference values of PLT count obtained in the study were lower than reference values that are currently used in most laboratories (150,000–450,000/μl) in India. CONCLUSION: Based on our results, we are of the opinion that the PLT count cutoffs for repeat plateletpheresis donation may need to be revised downwards for our country which would also mitigate the scarcity of apheresis donors.
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A study to assess the relationship between donor uric acid levels and supernatant hemolysis in stored packed red blood cell units p. 251
Himanshu Kumar Singh, Amit Kumar Biswas, Joseph Philip, Neerja Kushwaha, Bhasker Mukherjee, Ajay K Baranwal
DOI:10.4103/ajts.ajts_61_21  
BACKGROUND: Most of the red blood cell (RBC) storage lesions can be attributed to oxidative stress encountered by the RBCs throughout the duration of their storage. Various donor variables at the time of donation may be responsible for the total antioxidant capacity of the supernatant and thus, the “storability” and the magnitude of development of these RBC storage lesions. It is known that uric acid (UA) is responsible for more than 60% of the TAC of the blood. This study aims to explore the relationship between donor UA levels and the difference in percentage hemolysis, an important RBC storage lesion, on day 1 and day 21, in stored packed RBCs (PRBCs) units. MATERIALS AND METHODS: The serum UA of 100 healthy voluntary male blood donors was estimated at the time of blood donation. The percentage hemolysis in the supernatant of the leukoreduced citrate phosphate dextrose/saline–adenine–glucose–mannitol RBC units (n = 100) prepared from these donors was calculated on day 1 and day 21. The difference in percentage hemolysis between donors with high normal serum UA levels (>7 mg/dL) was compared to that of the donors with low normal serum UA levels (<5 mg/dL) to observe the effect of donor UA levels on the difference in percentage hemolysis. RESULTS: The mean of the differences in percentage hemolysis in the supernatant in low UA group (<5 mg/dL) was higher than the mean of the differences in percentage hemolysis in the supernatant in high UA group (>7 mg/dL) and this was statistically significant (P < 0.001). The donor serum UA level and difference in percentage hemolysis on day 21 and day 1 were found to be negatively co-related. CONCLUSION: Higher levels of serum UA of blood donors seem to have a protective effect on the stored PRBC units as shown in this study. Hence, the potential of UA as one of the constituents of RBC additive solutions might lead to the enhancement of the quality of stored PRBC units by decreasing the RBC storage lesions.
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UPDATE IN THE TRANSFUSION MEDICINE Top

Panagglutination on the indirect antiglobulin test... this is the challenge! p. 257
Styliani I Kokoris, Dimitrios Kalantzis, Dimitra Moschandreou, Konstantina Papaioannou, Elisavet Grouzi
DOI:10.4103/ajts.ajts_133_20  
Panagglutination on the indirect antiglobulin test is one of the most challenging dilemmas of pretransfusion testing. It occurs when patient sera react with all red blood cells tested, that is, with both screening and identification panel cells. Two main questions must be answered. The first is to determine whether panagglutination results from the presence of autoantibody and/or alloantibody (single alloantibody or multiple alloantibodies or antibody to high-incidence antigen). The second problem is to detect the possible concomitant presence of clinically significant alloantibodies masked by panagglutination. The purpose of this mini-review is to describe the situations that can cause panagglutination and to develop algorithms which can resolve the problem. The two main points in the evaluation of panagglutination involve the assessment of the intensity of reactivity with the reagent red cells used and whether the autocontrol is positive or not. It is imperative to understand the laboratory results and the techniques available that guide the investigative process.
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CASE REPORTS Top

Report of plasma exchange in a rare case of association of myasthenia gravis with thymoma and parathyroid adenoma p. 263
Nippun Prinja, Garima Siwach, Sarath Aleti, Rekha Hans, Aastha Takkar, Sahil Mehta, Ratti R Sharma, Vivek Lal
DOI:10.4103/ajts.ajts_84_21  
Myasthenia with thymoma and parathyroid adenoma is a rare presentation. Very few cases have been reported of this association without much role of plasma exchange in these patients. Here, we present our experience of plasma exchange in this rare clinical entity.
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Sickle cells are not necessarily protective against falciparum- A case report p. 266
Jess Elizabeth Rasalam, Snehil Kumar, K Sujith, NA Fouzia, Arpana Palle, Dolly Daniel, Sukesh Chandran Nair
DOI:10.4103/ajts.ajts_122_21  
The relation between sickle cell disease (SCD) and malaria is captivating where sickling of the infected red blood cells (RBCs) causes premature hemolysis and parasite death. Although patients with sickle cell trait are relatively protected, malaria can often lead to marked anemia in them due to hemolysis. We report an unusual case of a child with homozygous SCD presenting with falciparum malaria and had hyper parasitemia and severe anemia which completely resolved following treatment. Clinical suspicion in our case arose considering the endemic nature of malaria in our country. The two overlapping injuries to spleen reduced the clearance of parasites by the spleen as evidenced by high parasite load. Our case report reinforces malaria as a cause of clinical worsening of SCD and highlights the importance of a multifactorial approach in the management of worsening anemia in SCD.
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Management of Bombay Rh negative with clinically significant anti-S for CABG surgery p. 269
S Anuragaa, Dibyajyoti Sahoo, Pragya Silwal, Abhishekh Basavarajegowda, Esha Toora
DOI:10.4103/ajts.ajts_180_21  
The Bombay Rh D negative is the rarest of the rare in blood groups. A 65-year-old male patient with coronary artery disease was admitted for CABG. During grouping, forward showed no agglutination in A, B, D, and H, and reverse showed agglutination in A, B, and O cell. The blood group was confirmed as Bombay Rh D negative. Four units of PRBC was requested for the surgery as it was cardiothoracic surgery. We checked our inventory and rare donor list for Bombay-negative blood. Acute normovolemic hemodilution was done for 2 units preoperatively with saline replacement. Autologous platelet apheresis was done for this patient. During routine cross-match, one unit was incompatible. The patient had naturally occurring anti-S, which was reactive at 37°C and clinically significant. A total of 4 PRBC (Packed Red Blood Cell), 1 Single Donor Platelet (SDP), 12 Fresh Frozen Plasma (FFP), and 9 cryoprecipitate were transfused throughout the hospital stay. The patient was Bombay Rh negative with anti-S with major surgery, which was re-explored twice; the patient was managed successfully in spite of all these difficulties with cooperation from different blood banks from all over India.
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ABO-incompatible granulocyte transfusion: Is ABO subgroup a barrier? p. 273
Soma Agrawal, Mohit Chowdhry, Uday Kumar Thakur
DOI:10.4103/ajts.ajts_8_22  
Granulocyte transfusion (GTx) is an efficient and compelling treatment option for patients with neutropenia following hematopoietic stem cell transplant. The donor pool for granulocyte harvest is limited to close friends and family members and the donors accepted are often of the same ABO Rh type. We report a case of ABO-incompatible prophylactic GTx, in a case of acute myeloblastic leukemia. Postcollection processing of the granulocyte product was done to reduce the red blood cell volume to <5 ml, making it safe for transfusion. The transfusion was successful in stabilizing the total leukocyte counts in the patient. The patient was monitored, and there were no adverse reactions posttransfusion.
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A case of naturally occurring anti-Dia antibody in a young man p. 276
Salfarina Iberahim, Noor Haslina Mohd Noor, Mohd Nazri Hassan, Rosnah Bahar, Shafini Mohdmed Yusoff, Marini Ramli, Wan Suriana Wan Abdul Rahman, Zefarina Zulkafli, Marne Abdullah, Ho Sook Fong, Tengku Muzaffar Tengku Mohamed Shihabudin, Hisham Atan Edinur, Norul Hajar Che Ghazali
DOI:10.4103/ajts.ajts_136_21  
The Diego (Di) blood group system comprises 22 antigens located on the band 3 protein, most of which are low-prevalence antigens. The majority of antibodies to Diego system antigens were of clinically insignificant; however anti-Dia, -Dib, -Wra, -ELO and-DISK may cause hemolytic disease of the fetus and newborn (HDFN) and transfusion reaction. We reported a case of naturally occurring of anti-Dia in a young man who presented to our hospital for wound debridement of fingers injury. His serological results were suggestive of anti-Dia antibody, and his molecular blood group showed he has Di (a-b+) antigen. Anti-Dia may be clinically significant. It can cause mild-to-severe HDFN, but there are only infrequent reports of it being clearly implicated in a hemolytic transfusion reaction. We suggest the need for reagent red blood cell panels to include Dia antigen-positive cells in antibody identification tests for our populations.
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Direct antibody test negative autoimmune hemolytic anemia with pulmonary tuberculosis: A diagnostic challenge p. 280
Agarwal Sheetal, Singh Ruby, P Singh Dhirendra, Gupta Anubhav, Nandan Devki
DOI:10.4103/ajts.AJTS_75_20  
Tuberculosis (TB) has varied manifestations, but autoimmune hemolytic anemia (AIHA) due to TB is rare. Direct antibody test (DAT) or Coombs negative AIHA is also rare. We report a case of a 14-year-old boy who presented with hemolytic anemia and pneumonia. The Coombs test was repeatedly negative. After ruling out the possible infectious and noninfectious causes by extensive investigations, he was diagnosed as DAT-negative AIHA by monospecific antibody test with 4°C low ionic strength saline washes and column agglutination method which revealed the presence of IgG-2+ antibodies. Bronchoalveolar lavage fluid for acid-fast bacilli and gene Xpert was also positive. It is important to recognize TB as a cause of AIHA in South Asian countries where its incidence is high.
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BRIEF COMMUNICATION Top

Efficacy of cascade plasmapheresis in comparison with conventional therapeutic plasma exchange for relapsed atypical hemolytic uremic syndrome: A case report p. 283
Ashwinkumar Vaidya, Isha Polavarapu, Ravindra Attur Prabhu, Ganesh Mohan, Shamee Shastry, PA Prethika
DOI:10.4103/ajts.ajts_121_21  
Atypical hemolytic uremic syndrome (aHUS) is a rare and life-threatening disease that is associated with high mortality and morbidity. The incidence of aHUS is about 1 or 2 cases per 1,000,000 per year. Etiology can be either familial or sporadic. The pathogenesis of aHUS involves dysregulation of the alternative complement pathway, with predisposing mutations in complement genes. aHUS has a poor prognosis and a gradual or a relapsing (30%–86%) clinical course. The disease may present at any age but is mostly seen in children and young adults. Therapeutic plasma exchange (TPE) is one of the primary modalities of treatment in aHUS. This report presents the utilization of cascade plasmapheresis and its advantages over TPE in a patient with relapsed aHUS. There was a 73% decrement in antifactor H antibody levels following cascade plasmapheresis.
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LETTER TO THE EDITOR Top

The prevalence of Dia, Mia, and Mur antigens among Malaysians and Indians p. 287
Isha Polavarapu, Shamee Shastry
DOI:10.4103/ajts.ajts_5_21  
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2006 - Asian Journal of Transfusion Science | Published by Wolters Kluwer - Medknow
Online since 10th November, 2006