Influence of immunohematological markers on severity of in vivo hemolysis in human warm autoimmune haemolytic anemia.

Autoantibody production in autoimmune haemolytic anemia (AIHA) is the result of the loss of self-immunological tolerance of the host. Here we investigated the various immunohematological markers that may influence the severity of in vivo hemolysis in warm AIHA (WAIHA). Complete direct antiglobulin test (DAT) evaluation and immunohematological characterization were performed in 247 patients of WAIHA following departmental protocols. Clinical and laboratory details of patients were obtained from patient file. The median age of WAIHA patients was 47 years with a female preponderance. Lymphoproliferative diseases were the major underlying causes of secondary WAIHA. The mean haemoglobin (Hb) and reticulocyte count (Retic) were 6.43 gm/dL and 7.58% respectively. Single autoantibody bound to red cells was investigated in 151 patients. The main IgG subclass was IgG1. Multiple autoantibodies like IgG+ C, IgG+IgA and IgG+IgA+C were found in 87 (35.2%) patients. Free autoantibodies were observed in 112 patients with a median indirect antiglobulin test (IAT) reactivity of 2+. Derangement of haematological and biochemical values was statistically significant with increase in DAT reactivity, presence of multiple autoantibodies on red cells, coating of red cells by IgG3 or multiple IgG subclass, higher DAT dilution and increasing IAT reactivity. We conclude that several important but simple immunohematological parameters may influence the degree of in vivo hemolysis in WAIHA. Since a set of common haematological and biochemical test determines the severity of in vivo hemolysis therefore a comprehensive clinical and immunohematological evaluation is advisable for a correct diagnostic and therapeutic workup of WAIHA.

Kinetics of Covid-19 antibodies in terms of titre and duration among healthcare workers: A longitudinal study.

Background Most individuals with Covid-19 infection develop antibodies specific to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, the dynamics of these antibodies is variable and not well-studied. We aimed to determine the titres of naturally acquired antibodies over a 12-week follow-up. Methods We recruited healthcare workers who had tested positive on a specific quantitative reverse transcription-polymerase chain reaction (qRT-PCR) for SARS-CoV-2, and then tested for the presence of immunoglobulin G (IgG) antibody against the same virus at baseline and again at 6 and 12 weeks. The antibody titre was determined by a semi-quantitative assay based on signal/cut-off ratio. Healthcare workers with antibody positivity were divided into those with high titre (ratio ≥12) and low titre (<12). Their demographic details and risk factors were surveyed through a Google form and analysed in relation to the antibody titres at three time-points. Results Of the 286 healthcare workers, 10.48% had high antibody titres. Healthcare workers who had tested positive by qRT-PCR and those who had received the Bacille Calmette-Guérin (BCG) vaccination or other immune-boosters had a higher frequency of high antibody titres. While there was a significant decline in antibody titres at 6 and 12 weeks, 87.46% of individuals positive for IgG antibody persisted to have the antibody even at 12 weeks. Conclusion Healthcare workers who tested positive for SARS-CoV-2 on qRT-PCR had a high positivity for the specific antibody, which continued to express in them even at 12 weeks. Further follow-up is likely to enhance our understanding of antibody kinetics following SARS-CoV-2 infection.

Seroprevalence of COVID-19 Amongst Health Care Workers in a Tertiary Care Hospital of a Metropolitan City from India.

BACKGROUND: Seroprevalence studies for COVID-19 evaluate the extent of undetected transmission in a defined community, with special significance among health care workers (HCW) owing to their greater exposure and potential to transmit. METHODS: A total of 1122 HCW (approximately 25% of the employees) of a large tertiary care hospital in India were recruited for this cross-sectional study. COVID PCR-positive HCW were excluded. Based on their risk-assessment, participants were grouped into three categories. A questionnaire was administered and they were tested for SARS-CoV-2-IgG antibodies using the chemiluminescence. RESULTS: The overall seroprevalence among workers was 11.94%, which included 19.85% in COVID units, 11.09% in non-COVID units, and 8% in administrative workers (p=0.007). Antibody prevalence was highest in the department of gastroenterology (11.94%), followed by oncology (10.53%), pathology (10.26%), emergency medicine (7.84%) and critical care medicine (7%). Housekeeping staff, food and beverage staff, lab assistants and technicians had higher seroprevalence rate than doctors and nurses (p < 0.0001). HCW with a history of BCG vaccination in childhood and those who received an adequate prophylactic dose of hydroxychloroquine (HCQ) had a lower seroprevalence as compared to those who did not (7.31% vs. 16.8% and 1.30% vs. 11.25% respectively). CONCLUSION: BCG vaccination, HCQ prophylaxis, and the job profile influence the seroprevalence rate in HCW. Seroprevalence rate and follow-up evaluation of its durability may help hospitals to triage their staff at risk, rationalize their placement, prioritize the use of PPE, thereby potentially reducing the risk.

Managing uncontrolled postsplenectomy reactive thrombocytosis in idiopathic thrombocytopenic purpura: role of thrombocytapheresis.

Reactive thrombocytosis occurs in response to infection, trauma, or surgery. Splenectomy alone accounts for 19% of all possible causes of extreme thrombocytosis. We performed thrombocytapheresis in a young lady with chronic idiopathic thrombocytopenic purpura (ITP) who developed postsplenectomy reactive thrombocytosis. Her post splenectomy platelet count was 227 × 10(6)/ml which elevated to 1623 × 10(6)/ml on the 7th postoperative day. A single thrombocytapheresis procedure reduced her platelet to 403 × 10(6)/ml. She was discharged on the 10th postoperative day and then maintained a count of 204-238 × 10(6)/ml with aspirin. Thrombocytapheresis reduces the platelet count rapidly in thrombocytosis and prevents patients from having thrombotic events. However, such procedures should be performed very meticulously to ensure patient safety.

Era of blood component therapy: time for mandatory pre-donation platelet count for maximizing donor safety and optimizing quality of platelets.

Blood bank regulatory agencies including the Drug and Cosmetics Act (DCA) of India do not mandate a predonation platelet count in whole blood donation. Mandating such practice will definitely optimize the quality of random donor platelets (RDP) in terms of platelet yield and patient therapeutic benefit. We observed poor platelet yield in RDP concentrates prepared at our center with a significant number not meeting the DCA guideline of ≥ 4.5 × 10(10) per bag processed from 450 ml of whole blood. Therefore we planned this study to evaluate the pre-donation hematological values in our blood donor population and effect of these values on the quality of platelet concentrates. The prospective study included 221 blood donors eligible for donating 450 ml of whole blood (WB). Following the departmental standard operating procedure (SOP) RDPs were prepared using the 'Top & Bottom' quadruple bag system and automated component extractor. Quality of RDP was assessed as per departmental protocol. All results were recorded and subsequently transcribed to SPSS working sheet. A significant (p<0.001) decrement of donor blood counts has been observed after WB donation. Mean donor Hb and platelets reduced by 0.72 g/dl and 22.1 × 10(6)/ml respectively. Quality of RDPs in terms of platelet yield was significantly better (p<0.001) when donor platelet count was >200 × 10(6)/ml. Although platelet yield significantly correlated with the donor platelet count however quality of RDPs in terms of red cell contamination showed no correlation with the donor hematocrit. Platelet yield in random donor platelets is a concern in Eastern India. A platelet yield of 4.5 × 10(10) per bag as mandated by the DCA of India was only achieved when the donor platelet count was >200 × 10(6)/ml. Posttransfusion platelet recovery (PPR) was unsatisfactory in the transfused patient. Introduction of pre-donation platelet count in whole blood donation will maximize donor safety and optimize patient platelet transfusion management.

Induction of thrombotic thrombocytopenic purpura by dengue virus infection in a critical patient: Role of therapeutic plasma exchange.

Thrombotic thrombocytopenic purpura (TTP) secondary to dengue fever is a very unusual occurrence. Both conditions are fatal and can result in significant mortality and morbidity if left untreated. In this case, we present a young lady who suffered dengue fever followed by microangiopathic anemia, thrombocytopenic purpura, and altered sensorium. Investigations revealed microangiopathic hemolysis; there was no evidence of disseminated intravascular coagulation. As soon as, we diagnosed the patient as having TTP, we treated her with serial plasma exchange therapy, steroids, and monoclonal antibodies such as rituximab. The patient responded very well to the treatment and completely recovered from neurological symptoms and laboratory parameters also normalized. Hence, timely diagnosis and starting appropriate treatment immediately are key factors for successful outcomes.

Real-world data from India on clinical practices in the management of autoimmune haemolytic anaemia: A survey-based cross-sectional assessment.

BACKGROUND: Autoimmune haemolytic anaemia (AIHA) is a decompensated acquired haemolysis caused by the host's immune system acting against its own red cell antigens. The aim of this national survey is to capture real-world data of clinical practices in AIHA by collecting responses from clinical haematologists across India. METHODOLOGY: In this cross-sectional study, a structured, 26-question online survey was conducted in India by few members of the special interest group in immunohaematology between January and March, 2022. The final survey consisted of questions covering place of work, amount of AIHA cases being evaluated by the haematologist over preceding years, basic demographic, clinical and laboratory features of the patients being treated under them etc. Descriptive statistical analysis was performed during the assessment. RESULTS: The survey response rate was 48.2% (53/110), 69.8% (37/53) have diagnosed and managed more than 10 AIHA cases in the last 3 years with a female preponderance. There was considerable variability in response. While 56.6% (30/53) of respondents do have the access to the facilities to subtype AIHA cases; 32.1% (17/53) of clinicians would prefer administering high dose steroids for 6 weeks or more in non-responding patients, and only 45.3% (24/53) would assess the risks of thrombosis in AIHA. There is unanimous agreement among the participants that health-related quality of life should be taken into consideration in patients and the need for a national registry of patients with AIHA in India. CONCLUSION: The current national survey showed that some aspects of AIHA management were consistent; others were less so, but also significant variations were observed in certain clinical practices, where the evidence base is limited. A joint effort is needed to establish a national patient registry by including both clinical haematologists and transfusion medicine specialists which could potentially standardise AIHA management and future research in India.

Application of flow cytometry in transfusion medicine: The Sanjay Gandhi Post Graduate Institute of Medical Sciences, India experience.

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.

Alterations in blood component utilization in a tertiary care hospital in eastern India in the COVID-19 pandemic.

BACKGROUND: With shortages of blood globally in the COVID-19 pandemic, many hospitals and blood centers have successfully implemented blood conservation strategies and strategies to maintain blood donations and blood inventory. Here, we reported our experience of utilization of blood components for transfusion in the current COVID-19 pandemic and discussed the patient blood management (PBM) methods and importance of judicious blood usage in any pandemic. MATERIALS AND METHODS: The retrospective study of 8 months was divided into pre-COVID and COVID periods. Parameters that included number of blood requisitions, blood components reserved and transfused, clinical speciality wise blood usage in both the periods were analysed. Blood utilization indices were computed using recommended equations. RESULTS: We observed an unusual pattern of blood utilization with significant disruption on blood supply and demand chain in this pandemic. A mean drop of 50.7% blood requisitions was observed in the COVID period. The period observed transfusion of 3608 units of blood components with a mean drop of 49.7% when compared to pre-COVID period. Mean drop of 46.6%, 54.4%, 53.4% and 52.7% were observed in packed red blood cell, fresh frozen plasma, platelet concentrate and cryoprecipitate transfusions in the COVID period. Blood component utilization was significantly reduced in most clinical specialities. The mean cross match to transfusion ratio in the COVID period was 1.28 with a mean increase in transfusion probability and transfusion index by 18.2% and 22.2% respectively. CONCLUSION: Although a constant transfusion support was needed in few clinical specialities throughout the pandemic, strict adherence to PBM protocols and practice of first in, first out method of blood dispensing helped the blood centre to support all patients needing blood transfusion.

Seroprevalence of anti-severe acute respiratory syndrome coronavirus 2 antibody among healthy blood donors in a hospital-based blood center in Eastern India during the COVID-19 pandemic.

BACKGROUND: Few international studies have reported the prevalence of anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies in healthy, asymptomatic blood donors. These findings have definitely raised queries regarding blood safety and transfusion-transmitted coronavirus disease (COVID)-19. We conducted this first anti-SARS-CoV-2 seroprevalence survey among the healthy blood donors in Eastern India. MATERIALS AND METHODS: The study included 611 healthy blood donors who donated whole blood (WB) in our blood center. For detailed analysis, social and demographic details of all donors like gender, age, weight, occupation, and place of residence were included. Donor eligibility criteria for WB donation were followed as per existing national guidelines. Residual serum samples leftover after screening the mandatory infectious markers were tested for the presence of anti-SARS-CoV-2 IgG directed against domain S1 and S2 of the SARS-CoV-2 spike protein using automated enhanced chemiluminescence technology following the manufacturer's instructions. RESULTS: The mean overall seroprevalence of anti-SARS-CoV-2 antibody in blood donors was observed to be 4.4% (95% confidence interval 3.8-4.9) with a monthly increasing trend. Seroprevalence adjusted for sensitivity and specificity of the assay was 4.1%. The mean S/Co values of reactive donor samples were observed to be 2.99 and 3.42 in June and July 2020, respectively (P = 0.013). No significant variation in seroprevalence rate was observed among donor variables like donor age, gender, profession, and educational qualification. A higher significant prevalence of antibody was observed among voluntary donors and donors residing in suburban areas (P < 0.05). Among the ABO blood groups, no statistical significance of seroprevalences was observed among the various ABO blood groups. CONCLUSION: We conclude that despite many limitations in the current study, we found 4.4% seroprevalence of anti-SARS-CoV-2 antibody in the asymptomatic, healthy, epidemiologically, and medically screened blood donors. These data are definitely the tip of an iceberg and signify much higher seroprevalence in the normal population and indicate that protective measures like masking and social distancing should remain implemented for a long term.

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.

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.

"Auto-anti-A1" in a healthy young blood donor: A rare cause of ABO discrepancy.

"Auto-anti-A1" has been sparsely discussed in the literature. Only few workers in the past depicted such antibody in transfused and nontransfused patients. The current case is probably the first example of auto-anti-A1 in a healthy young blood donor who was typed as ABO Group "A1B." The cold reacting autoantibody in the donor was serologically characterized in details and was found to be nonhemolytic. ABO discrepancy was resolved and the donor was finally typed as "A1B Negative." Therefore, we concluded that auto-anti-A1 may be a rare cause of ABO discrepancy and its resolution is essential to confirm blood group and subsequent blood transfusion management.

Plateletpheresis in the Era of Automation: Optimizing Donor Safety and Product Quality Using Modern Apheresis Instruments.

The increases in major surgeries, transplantations and speciality clinics have significantly increased the utilization of platelet concentrates including single donor platelets (SDP). The advantages of SDP or apheresis platelet have been discussed elaborately by previous authors as compared to random donor platelets. Here we share our experiences of plateletpheresis procedures using the modern apheresis machines with regards to product quality and donor safety. This study included 3016 procedures of plateletpheresis (1397 on Amicus and 1619 on Trima accel cell separators) on eligible donors using recommended apheresis kits. A target yield of 3 × 1011 was set as the end point of each procedure. Donor details, procedure details and donor adverse reactions if any were documented. Statistical analysis was done using the SPSS statistical package (version 13, USA). Of the total 6276 donors screened 2049 (32.6%) were deferred due to various reasons. Out of remaining 4227 eligible donors; 3016 (71.4%) underwent plateletphereis procedures based on the requirement of SDP by the patients. Mean pre-procedure platelet count and hematocrit in donors were 188.3 × 106/mL and 41.7% respectively. Mean procedure time in Amicus (76.6 min) was significantly more than the Trima accel (64.3 min) (p = 0.02). Platelet yield by Trima accel and Amicus was 2.96 × 1011 and 3.08 × 1011 respectively (p = 0.061). A total of 40 donors (1.33%) suffered adverse effect during or after apheresis procedures. While the modern plateletpheresis devices are both donor and user friendly at the same time they provide quality product consistently in lesser time.

An insight into the utilization of allogenic blood transfusion and factors affecting blood transfusion in total knee replacement surgery in a tertiary care hospital in Eastern India.

BACKGROUND: Significant blood loss and requirement of allogenic blood transfusion during or after total knee replacement (TKR) have been reported. Incidence of blood transfusion in TKR is highly variable and depends on several factors. We investigated the blood utilization in patients undergoing TKR in our hospital and depicted the important risk factors that determine the need of allogenic blood transfusion in primary unilateral TKR. MATERIALS AND METHODS: The study included 1241 consecutive patients undergoing primary unilateral total knee arthroplasty. All the surgeries were performed by a single surgical team of orthopedists following standard procedure. Patient and disease details were obtained from patient file and hospital information system. Compatibility test was performed in blood bank before blood reservation following mandatory guidelines. Details of test, blood issue, and blood transfusion were documented in the blood bank. RESULTS: Of 1241 enrolled patients, 1069 (86.2%) were female. The median age of patients was 66 years with mean preoperative hemoglobin of 9.9 g/dL. Allogenic blood transfused was needed in 223 (17.9%) patients. Diabetes mellitus, hypertension, thyroid disorders, and chronic heart diseases were the major comorbid conditions. Risk factors such as gender, American Society of Anesthesiologists score, preoperative hemoglobin, and intraoperative and postoperative blood losses were significantly associated with blood transfusion. CONCLUSION: The risk factors determining blood transfusion in TKR vary between studies, however, all centers should establish standard operating procedures describing the surgical procedure and transfusion support in TKR. In addition, each center may develop specific blood management strategy to rationalize blood transfusion in TKR and overall successful care in TKR.

Immunohematological and Clinical Characterization of Complement and Non-Complement Associated Warm Autoimmune Haemolytic Anemia and Risk Factors Predicting their Occurrences.

Antigen - antibody complexes on heavily coated red cells in Warm autoimmune haemolytic anemia (WAIHA) often activates the complement pathway and red cells bound C3 complement component are encountered in complement associated WAIHA (CWAIHA). Patients belonging to CWAIHA and non-complement associated WAIHA (NCWAIHA) may demographically, clinically and immunohematologically behave differently therefore we planned to study the clinical and immunohematological characteristics of CWAIHA and NCWAIHA with emphasis to various potential factors associated with CWAIHA. The prospective study included 229 patients of WAIHA. Complete DAT evaluation was performed in all these patients. Details of patients and their hematological and biochemical parameters were obtained from patient file and Hospital Information System. In vivo hemolysis was documented as per the criteria established by previous workers. Statistical analysis was done using SPSS statistical package. Of the total 229 patients of WAIHA, 83 (36.2%) belonged to the complement associated WAIHA group. A total of 146 (63.8%) patients were females of which 43 (29.4%) had CWAIHA. The median age of WAIHA patients was 37 years. A total of 46 (56.1%) patients above age 40 years suffered from CWAIHA. Where secondary WAIHA was found in 121 (52.8%) patients; more than half (61.4%) with CWAIHA had underlying aetiology. Over 95% of patients in both categories presented with weakness and pallor. Strong DAT (> 2 +) was observed in 86.7% of CWAIHA patients. Factors like gender, age, aetiology and DAT IgG dilution were independent risk factors for CWAIHA. DAT remained positive even at the end of 10 months of successful treatment. We conclude that detailed characterization of WAIHA with particular emphasis to complement and non-complement associated WAIHA is essential to evaluate the disease characters, immunological behaviours, prognosis and therapeutic management. Moreover an understanding of the risk factors of CWAIHA will help physicians / hematologists and immunohematologists to manage WAIHA more prudently and solicitously.

Rationalizing blood transfusion in elective breast cancer surgery: Analyzing justification and economy.

BACKGROUND: Transfusion of allogeneic blood in breast cancer surgery is variable, and differences of transfusion incidence have been observed in the literature. Most hospital guidelines including ours dictate group and reserve policy of blood before breast surgery. Here, we aimed to audit the blood utilization in patients undergoing elective breast surgery in our hospital and thereby optimize the blood ordering schedule, economic burden, and loss of clinical resources. MATERIALS AND METHODS: The study included 478 breast cancer surgeries over a period of 6 years. Patient and disease details were obtained from patient file and hospital information system. Blood samples sent to blood bank were subjected to compatibility test and reserved. All transfusions were documented, and statistical analysis was done. RESULTS: Of the total 478 patients, most underwent wide local excision of the breast and modified radical mastectomy. A total of 16 patients received 71 units of blood and blood components in all categories of surgeries. Only 103 were younger women (≤40 years), with a mean age of 31 years. Nontransfused patients were significantly more than transfused ones (P < 0.05). Frequency of blood transfusion was more in young patients (4.9%). Seven (22.6%) of the total 31 Stage IV patients received blood transfusions. Frequency of blood transfusion was more in patients undergoing surgery after chemotherapy (8.8%). A significant loss of time and loss of revenue was observed. CONCLUSION: We conclude that routine compatibility test is not justified for all patients undergoing breast surgery. A more targeted approach is needed to reduce blood demand and associated cost to patient and blood transfusion services.

Clinical and serological characterization of autoimmune hemolytic anemia in a tertiary care hospital in North India.

Clinical and serological characterization of autoimmune hemolytic anemia (AIHA) helps in the diagnosis, management, and monitoring course of disease. In the present study, we serologically characterized the red-cell-bound autoantibodies in diagnosed AIHA patients with regards to antibody class, subclass, direct antiglobulin test (DAT) strength, and their correlation with in vivo hemolysis. A total of 157 samples were evaluated for DAT. Clinically and serologically, 43 of them were diagnosed as AIHA. Detailed serological characterization of autoantibodies was performed in these 43 patients using the gel technology. Hematological and biochemical parameters were obtained from the Hospital Information System. Polyspecific (immunoglobulin G (IgG) + C3) DAT-positive samples were tested for monospecific DAT (IgG, IgM, IgA, C3c, and C3d) and IgG subclass (IgG1 and IgG3). Thermal amplitude of autoantibodies was determined on eluates. Median age of the patients was 31 years (range, 12-70 years) with male to female ratio of 1:3.3. In 55.8% of patients, AIHA was secondary to an underlying disorder. Patients with strong reactive DAT had increased likelihood of hemolysis (p = 0.000). IgG was the solitary autoantibody coating the red cells in 72.1% of patients. Red cells coated with multiple immunoglobulins/complements and IgG subclass IgG1 and/or IgG3 were more susceptible to undergo hemolysis. Gel technology helps the immunohematologist to diagnose and serologically characterize AIHA patient with regard to red-cell-bound autoantibodies' class, subclass, and titer as these correlate with in vivo hemolysis.

Comparative study of automated plateletpheresis using five different apheresis systems in a tertiary care hospital.

Different types of cell separators are available nowadays based on either continuous or intermittent flow technology to meet the growing demands for single donor apheresis platelets. This prospective study compares the five machines used in our center with regard to procedure parameters, product quality and adverse effects on the donor. A total of 477 plateletpheresis on various machines were performed on eligible donors over a period of 28 months after taking informed consent. All procedures were performed following the departmental standard operating procedure (SOP) and manufacturer's instructions. All donor and procedure related details were obtained from the procedure register. Statistical analysis was done using the SPSS statistical package (version 12, USA). The median age, BSA and BMI of our plateletpheresis donors were calculated to be 29 years, 1.69 m(2) and 23.1 kg/m(2), respectively. Analyzing the parameters related to donor comfort such as donation time (DT), needle time (NT) and processing time (PT), the MCS machines were not "donor friendly" compared to Amicus and Fresenius. Platelet yield by Amicus was significantly higher as compared to other cell separators (p < 0.05). Plateletpheresis associated citrate toxicity was higher with the Amicus and MCS 3p and vasovagal side-effects was observed least with the CS 3000 machine. Though, quality of apheresis product in terms of yield is comparable with all the machines, there are differences in the systems with regard to donor safety, procedure time and donor retention.

Masquerading of mismatched blood transfusion by underlying autoimmune hemolytic anemia.

Mismatched blood transfusion due to immunohematological discrepancy is relatively uncommon and in most instances occurs due to Type IV blood group discrepancy which is the discrepancies between forward and reverse groupings. Here, we present a case of a 15-year-old girl with preexisting autoimmune hemolytic anemia (AIHA) who inadvertently received 3 units of wrongly matched packed red blood cell (PRBC), followed by severe intravascular hemolysis. On detailed immunohematological investigation, the patient was found to be autoimmunized and diagnosed with "mixed AIHA" and the patient's blood group was confirmed as "A" positive. Three units of group-specific "best match" PRBC was transfused under close observation without any adverse effect. This highlights the importance of carrying out both forward and reverse blood groupings to avoid mismatched blood transfusion.

Quality analysis of red cell and platelet concentrates obtained by the automated 'Top-and-Top' blood processing system in a developing country.

The efficacy of the widely practiced 'Top and Bottom' collection system has been established. We studied efficacy of our new 'Top-and-Top' automated blood component separation system with regards to quality analysis of red cell and platelet concentrates. At installation of machine 59 U of whole blood (WB) were used for its calibration and validation program. Optimum volume, leukocyte-platelet recovery and red cells loss in BC were adjusted as per recommended standards. WB showed mean volume of 510 ml and net hemoglobin content of 63 g/bag. BC recovered 91.7% and 62.7% of platelet and leukocytes, respectively, in a mean volume of 96 ml a hematocrit of 54% and mean platelet of 5.47 x 10(10) were observed in red cell and platelet concentrates, respectively. In many aspects quality of our products could not comply with the recommended European and American standards and this requires a close insight into the series of activities associated with WB collection, separation and quality control program.