Evaluation of chemiluminescence immunoassay as a screening test for syphilis on blood donor samples.

BACKGROUND AND OBJECTIVE: Global re-emergence of syphilis among blood donors necessitates novel diagnostic and prevention approaches that encourage timely intervention. Thus, the present study was planned to evaluate the efficiency of Chemiluminescence immunoassay (CLIA) as a screening test for syphilis. MATERIAL AND METHODS: This prospective cross-sectional observational study was conducted from October 2021 to September 2022. A total of 344 donors were enrolled by purposive sampling method, including additional 16 donors who were reactive by the Rapid plasma reagin test (RPR) during the study period. Data from three screening tests - RPR test, Treponema pallidum haemagglutination assay (TPHA) and CLIA for 360 blood donors were analysed. TPHA was considered the gold standard test. RESULTS: Of the total 360 samples tested, 21 (5.8 %) were reactive by the RPR test. Of these 21 RPR reactive samples, 19 (90.5 %) were reactive by both TPHA and CLIA, while 2 (9.5 %) RPR reactive samples were non-reactive by both TPHA and CLIA. Of the remaining 339 RPR non-reactive samples, 1 (0.3 %) sample was reactive by both TPHA and CLIA, and 1 (0.3 %) was reactive by CLIA alone. CLIA was found to have sensitivity and specificity of 100 % and 99.7 % and positive predictive value (PPV) and negative predictive values (NPV) of 95.2 % and 100 % respectively, while it was 95 %, 99.4 %, 90 %, and 99.7 %, respectively, with the RPR test. CONCLUSION: CLIA was found to have a higher sensitivity, specificity, PPV and NPV than the RPR test. Thus, CLIA can be an acceptable alternative for syphilis screening in blood donors.

Physiological and psychological stress response of blood donors during the blood donation process.

BACKGROUND AND OBJECTIVES: Blood donation can be a potentially stressful event, leading to the activation of an acute stress response. Knowing and identifying potential stressors could help in optimizing the donation experience. The present study aimed to measure the physiological and psychological stress changes before, during and after blood donation. MATERIALS AND METHODS: Physiological and psychological stress response was assessed in 70 blood donors. To evaluate physiological stress response, pulse rate, respiratory rate, blood pressure (BP), beat-to-beat BP and lead II electrocardiogram were recorded. Baroreflex sensitivity was calculated using the available software. Psychological stress response was assessed using the State-Trait Anxiety Inventory scale. RESULTS: A significant increase in systolic blood pressure, diastolic blood pressure and mean arterial pressure was observed in the pre-donation period (p < 0.001). Among the time-domain parameters, SDSD (standard deviation of differences between adjacent respiratory rate intervals) and RMSSD (root mean square of the successive differences) were significantly lower during the post-donation period (p < 0.005, p < 0.007, respectively). Among the frequency-domain parameters, LF nu (relative power of the low-frequency band in normalized units), HF nu (relative power of the high-frequency band in normalized units) and LF% (relative power of the low-frequency band in percentage) were significantly lower before donation compared to during donation (p < 0.001, p < 0.001 and p < 0.012, respectively). LF nu, LF% and LF/HF ratio were also significantly lower during donation compared to after donation (p < 0.05, p < 0.016 and p < 0.042, respectively). Baroreflex sensitivity was also statistically higher during the pre-donation period. State score was significantly higher among the blood donors during the pre-donation period. CONCLUSION: Physiological and psychological stress is experienced by blood donors during the pre-donation period. A pre-donation informative conversation should be carried out with each blood donor and potential stressors should be identified in each.

Association of blood donor characteristics and in vitro haemolysis of packed red blood cell concentrates during storage.

BACKGROUND AND OBJECTIVES: Blood donor variability can affect the storage properties of packed red blood cells (PRBCs). This study aimed to determine the association of donor characteristics with in vitro storage haemolysis of PRBCs. MATERIALS AND METHODS: In the prospective observational study, a total of 109 whole blood donors were enrolled using the purposive sampling method. A pre-donation sample was collected for haemoglobin (Hb) and serum uric acid (UA) levels. PRBC aliquots were tested for potassium, lactate dehydrogenase (LDH), Hb, haematocrit, plasma Hb and haemolysis on days 1, 21 and 35 of storage. The association of these parameters with donor age, sex, donation status, dietary pattern and body mass index was determined. RESULTS: Mean haemolysis was significantly higher in PRBCs from donors with UA levels ≤6 mg/dL than donors with UA levels >6 mg/dL on day 35 of storage (0.22 ± 0.11 vs. 0.18 ± 0.07, p = 0.03). Median plasma Hb (mg/L) was significantly higher in PRBCs from first-time donors on day 21 (586 vs. 509, p = 0.05) and day 35 (1507 vs. 1358, p = 0.02) of storage in comparison to frequent donors. Significantly higher mean potassium (p = 0.04 day 1; p = 0.02 day 21) and median LDH values (p = 0.02 day 1, p = 0.05 day 21) were observed in PRBCs from male donors. A statistically significant positive association was observed between donor UA and LDH levels of PRBCs on day 35 of storage (ß coefficient: 715.52, p-value: 0.003) on multiple regression analysis. CONCLUSION: In vitro haemolysis of PRBCs is affected by blood donor characteristics.

Association of blood donor characteristics with hemoglobin content in leukoreduced packed red blood cells.

BACKGROUND AND OBJECTIVES: A complex relationship exists between donor characteristics and red blood cell quality which remains partly explored. The present study aimed to determine the correlation of donor characteristics with the hemoglobin (Hb) content of leukoreduced packed red blood cells (PRBC). MATERIALS AND METHODS: This prospective cross-sectional study was conducted on 100 blood donors. A pre-donation sample was collected for hemoglobin and hematocrit estimation. Whole blood was collected in quintuple blood bags and packed red cells were prepared. Sample from each packed red cell unit was estimated for hemoglobin and hematocrit. The volume, total Hb, actual total Hb, volume and Hb lost during processing, mathematical total Hb and hematocrit of each PRBC unit was calculated using formulas. The donor characteristics were analysed for correlation with Hb content of PRBC. RESULTS: The mean age of the 100 donors enrolled in the study was 36.3 ± 9.9 years. Majority of the donors were vegetarian, non-alcoholic, non-smokers, and had a pre-donation hemoglobin level of more than 14 g/dl. The mean pre-donation Hb of the donors was 14.8 ± 1.5 g/dl. There was a strong positive correlation of donor pre-donation hemoglobin with total Hb (r = 1.000, p = 0.000), actual Hb (r = 0.518, p = 0.000) and mathematical hemoglobin (r = 0.951, p = 0.000) using the Pearson correlation test. A strong positive correlation was observed between the total and actual hemoglobin (r = 0.518, p = 0.000) of the units. There was no association of other donor characteristics with Hb content of leukoreduced PRBC. CONCLUSION: Donor pre-donation hemoglobin showed a strong positive correlation with the actual hemoglobin content of leukoreduced packed red blood cells.

Association of blood donor characteristics with in vitro platelet storage properties.

BACKGROUND AND OBJECTIVE: The study was planned to determine the association of blood donor characteristics with in vitro quality of platelets. MATERIAL AND METHODS: In the prospective observational study, a total of 85 male whole blood donors in the age group of 18-30 and 45-65 years were enrolled using purposive sampling method. Serum total cholesterol, glycosylated hemoglobin (HbA1c), and LDH levels were performed on donor pre-donation sample. Buffy coat platelet concentrates were prepared from 450 mL quadruple blood bags. Samples from platelets were taken on day one and five of storage and biochemical properties were observed. RESULTS: Median MPV was higher in platelets from older blood donors on day five (9.8 vs 9.4, p = 0.037). Median LDH levels were also higher in platelets on day one and five from older donors (Day one: 204.5 vs 147, p = <0.000; day five: 278 vs 224, p = 0.001 respectively). Platelets from donors with high HbA1c levels had lower median pH (Day one: 7.31 vs 7.37, p = 0.024) and higher median glucose levels on day one of storage (Day one: 358 vs 311, p = 0.001). Higher median lactate levels throughout the storage period were also seen in platelets from donors with higher HbA1c levels (Day one: 7 vs 5.7, p = 0.037; Day five: 16 vs 12.2, p = 0.032). Glucose consumption (108 vs 66, p = 0.025) and lactate production (9 vs 6.4, p = 0.019) was higher in platelets from donors with higher HbA1c levels. CONCLUSION: In vitro platelet storage properties are affected by blood donor characteristics.

Revisiting the impact of serial therapeutic phlebotomy in polycythaemia on laboratory and clinical parameters using a fixed interval and fixed volume protocol.

OBJECTIVES: Therapeutic phlebotomy allows for a controlled and gradual decrease in red cell mass leading to improved blood flow and symptomatic relief in polycythaemia. The present study was aimed to determine the impact of serial fixed volume and fixed interval therapeutic phlebotomy protocol on the laboratory and clinical parameters in patients of polycythaemia. MATERIAL AND METHODS: This prospective longitudinal study was conducted over 18 months. The desired haematocrit for polycythemia vera and secondary polycythemia was 45% and 52% respectively. A fixed volume of 350 ml phlebotomy was performed every-three days till the achievement of desired haematocrit. Complete blood count was performed before and after each procedure and iron studies were done at the time of enrolment and after the achievement of desired haematocrit. Post-procedure symptomatic relief was assessed by a 10-point visual analogue scale (VAS). RESULTS: Of the 29 patients enrolled in the study, 3 patients were lost to follow up and data of 26 patients was analyzed. Mean Hb declined from 17.84 ± 1.88 gdL-1 to 14.67 ± 1.14 gdL-1 (p < 0.001) and mean haematocrit decreased from a baseline of 57.11 ± 5.47% to 46.27 ± 3.763% (p < 0.001) upon achievement of desired haematocrit. There was a significant decline in serum iron from the baseline of 132.85 ± 94.136 µg dL-1 to 69.41 ± 58.643 µg dL-1 at desired haematocrit. A significant change in VAS score of almost all clinical parameters was observed. Post phlebotomy hematocrit correlated negatively with the number of procedures (p = 0.015). CONCLUSION: Our protocol yielded rapid and marked improvement in patients of primary and secondary polycythemia with minimal adverse events and significant amelioration of clinical parameters.

Exploring the unseen effect of COVID 19 pandemic on blood transfusion services in a tertiary care centre.

BACKGROUND: The coronavirus pandemic confronted blood transfusion services with major challenges. The present study was conducted to explore the effect of the COVID-19 pandemic on blood transfusion services including seroprevalence of transfusion-transmitted infections. MATERIAL AND METHODS: A retrospective cross-sectional study was conducted and data on blood donation, utilization, camps, plateletpheresis and seroprevalence of transfusion-transmitted infections (TTI) was retrieved from software from March to September 2020 and 2021 and compared with corresponding time periods of three preceding non-pandemic years. RESULTS: There was a decline of 53.79% and 34.4% in blood donations in 2020 and 2021 respectively with a significant reduction in voluntary donations from 91.8% in the pre-pandemic period to 72.2% in 2020 and 77.7% in 2021. Replacement donors increased by 60.81% and 72.89% in 2020 and 2021 respectively. There was a decline of 48.4% in the number of plateletpheresis procedures in 2020 which increased in 2021 during the dengue outbreak. The decline in total blood donations and issue of packed red blood cells was statistically significant but supply and demand were balanced with no deficit. TTI seroprevalence increased from 1.01% to 1.49%(p<0.001) and 1.51%(p<0.001) in 2020 and 2021 respectively. Replacement donors showed a significantly higher TTI prevalence as compared to voluntary donors(p<0.001). A significant increase in prevalence was observed for Syphilis (0.4%) in 2020 and HBsAg (0.54%), HCV(0.63%) and syphilis (0.25%) in 2021. CONCLUSION: The potential consequences of the COVID-19 pandemic on blood safety cannot be undermined. Developing a strong database of regular voluntary donors can be instrumental in dealing with future waves and surges in infections.

Efficacy of packed red blood cell transfusions based on weight versus formula in thalassemic children: An open-label randomized control trial.

BACKGROUND: Protocols for transfusion therapy in transfusion-dependent thalassemia (TDT) children differ among various medical centers. In India, most centers consider only the patient's weight while calculating the volume of packed red blood cells (PRBCs) to be transfused. This study aimed to compare the efficacy of PRBC transfusions of different volumes calculated either by weight or by a formula using weight and pretransfusion hemoglobin of patient and hematocrit of PRBC. STUDY DESIGN AND METHODS: Sixty TDT patients in the age group of 3-9 years were enrolled and randomly allocated to two groups. Group A received PRBC transfusion volume based on the patient's weight, and Group B received PRBC volume calculated using a formula for 6 months. RESULTS: Average pretransfusion hemoglobin in Group A and Group B (9 ± 0.4 vs. 8.9 ± 0.4 g/dl) was not significantly different (p = .353). Although the average number of visits in 6 months was less for Group A compared to Group B (7 ± 1 vs. 8 ± 1; p = .001); the average volume transfused per visit was more (351 ± 78 vs. 287 ± 68 ml; p = .003). The calculated average annual pure red cell requirement of the patients was 178 ml/kg/year for Group A and 154 ml/kg/year for Group B (p = .000). Total donor exposures were significantly lower in Group B than Group A (11 ± 3 vs. 14 ± 3; p = .006). CONCLUSION: The number of donor exposures and annual pure red cell requirement was significantly lower in the formula-based group. Transfusions based on formula are recommended in TDT patients.

Assessment of serum iron stores in regular plateletpheresis donors.

BACKGROUND: Iron deficiency anaemia is the most common nutritional deficiency disorder in the world. Iron deficiency is a potential complication in repeated apheresis donation. The present study was aimed to evaluate serum iron stores in regular plateletpheresis donors. MATERIALS AND METHODS: A total of 60 donors were included in this study, which included 30 regular plateletpheresis donors as cases and controls were 30 first time donors. The donor samples were collected before donation for complete hemogram, transfusion transmissible infections screening and serum iron, total iron binding capacity, percentage saturation of transferrin and serum ferritin. RESULTS: Out of 60 donors, more than half of the donors (56.6 %) had serum ferritin less than 30 ng/mL. Out of these 34 donors, 25 were from the case group and 9 donors in the control group. The median serum ferritin level in cases and controls was 11.86 ng/mL (Interquartile range 4.18-17.34 ng/mL) and 37.92 ng/mL (Interquartile range 27.87-86.20 ng/mL) respectively (p < 0.001). The mean serum iron in cases and controls was 71.23 ± 31.32 µg/dL and 93.53 ± 33.53 µg/dL respectively (p = 0.016). The mean percentage saturation in cases and controls was 20.09 ± 9.31 % and 26.26 ± 9.03 % respectively (p = 0.012). A significant decline in mean serum ferritin with increase in number of annual donations and decrease in donation interval was observed. DISCUSSION: Regular plateletpheresis donation may lead to depletion of iron stores and subclinical iron deficiency. Donors with high platelet count are more likely to exhibit iron deficiency. Periodic serum ferritin estimation in donors participating in regular plateletpheresis donation is warranted.

Challenges in management of atypical hemolytic uremic syndrome: Bottle neck in resource limited settings.

Treatment of atypical hemolytic uremic syndrome cases is challenging right from establishing correct and timely diagnosis to execution of management protocol. A seven-year-old male child from poor socioeconomic status was admitted with chief complaints of fever, 3 episodes of vomiting and passage of cola coloured urine. Based on clinical and laboratory findings, diagnosis was established. However, ADAMTS13 levels and genetic studies required for diagnosis could not be performed due to financial constraints and non-availability of these tests. TPE kits were arranged from charitable organizations. Six TPE procedures were performed using Cobe Spectra cell separator. Central venous catheter was placed in femoral vein. TPE kit was primed with compatible packed red blood cells before each procedure. Patient was non-cooperative and irritable in first three procedures and was sedated. A total of 1300ml plasma was exchanged in each procedure with group specific fresh frozen plasma. After second TPE procedure, patient started improving with decrease in plasma discoloration and periorbital edema. Renal function tests along with hematological parameters became normal after 6th TPE procedure. Patient was discharged in a stable condition. On follow up, C3 levels were normal with adequate platelet count and normal renal functions suggesting complete remission.

Utility of reticulocyte haemoglobin content and immature reticulocyte fraction in early diagnosis of latent iron deficiency in whole blood donors.

BACKGROUND AND OBJECTIVES: The present study was planned to assess the clinical utility of reticulocyte haemoglobin content (CHr) and immature reticulocyte fraction (IRF) in the early detection of latent iron deficiency in blood donors. MATERIALS AND METHODS: The prospective longitudinal observational study was conducted using the purposive sampling method. Written informed consent was obtained and donors were allocated into the first-time (FTD) and regular donor (RD) group. The enrolled blood donors (n = 205 in each group) were followed up for two subsequent whole blood donations. Haemoglobin (Hb), CHr, IRF and serum ferritin values were recorded at enrolment and two follow-ups. RESULTS: The sensitivity of CHr in detecting iron-deficient erythropoiesis (serum ferritin values ≤ 26 µg/dl) was 45% and 56.7%, specificity 96.7%, positive predictive value (PPV) 85.6% and 90.8% and negative predictive value (NPV) 80.1% and 78.7%, respectively in FTD and RD cohorts. The sensitivity of IRF was 45.1% and 44.8%, specificity 93.4% and 97.1%, PPV 74.8% and 90.4% and NPV 79.6% and 74.5%, respectively in both the cohorts. The sensitivity of CHr in detecting absent iron stores (serum ferritin values ≤ 15 µg/dl) was 66.2% and 74.4%, specificity 92% and 90.6%, PPV 56.7% and 68.7% and NPV 94.5% and 92.8% among FTD and RD cohort, respectively. The sensitivity of IRF was 72.7% and 65.3%, specificity 90.3% and 94.3%, PPV 54.4% and 76% and NPV 95.4% and 90.8%, respectively in both the cohorts. CONCLUSION: Reticulocyte hemoglobin content and IRF can be used along with complete blood count for early detection of iron deficiency in blood donors using the same blood sample at no extra cost.

Genome-wide association analysis of Mexican bread wheat landraces for resistance to yellow and stem rust.

Deploying under-utilized landraces in wheat breeding has been advocated to accelerate genetic gains in current era of genomics assisted breeding. Mexican bread wheat landraces (Creole wheats) represent an important resource for the discovery of novel alleles including disease resistance. A core set of 1,098 Mexican landraces was subjected to multi-location testing for rust diseases in India, Mexico and Kenya. The landrace core set showed a continuous variation for yellow (YR) and stem rust (SR) disease severity. Principal component analysis differentiated Mexican landraces into three groups based on their respective collection sites. Linkage disequilibrium (LD) decay varied from 10 to 32 Mb across chromosomes with an averge of 23Mb across whole genome. Genome-wide association analysis revealed marker-trait associations for YR resistance in India and Mexico as well as for SR resistance in Kenya. In addition, significant additive-additive interaction effects were observed for both YR and SR resistance including genomic regions on chromosomes 1BL and 3BS, which co-locate with pleiotropic genes Yr29/Lr46/Sr58/Pm39/Ltn2 and Sr2/Yr30/Lr27, respectively. Study reports novel genomic associations for YR (chromosomes 1AL, 2BS, and 3BL) and SR (chromosomes 2AL, 4DS, and 5DS). The novel findings in Creole wheat landraces can be efficiently utilized for the wheat genetic improvement.

Clinical Significance of Rare Maternal Anti Jka Antibody.

We hereby report a rare case of HDFN because of antibody to Kidd (Jk) blood group system-anti Jka. An EDTA sample of a baby along with mother's sample was received in the Department for Direct Antiglobulin Test (DAT) alongwith blood requisition for double volume exchange transfusion. On blood grouping, baby's and mother's blood group was found to be B Rh D positive. DAT with polyspecific anti human globulin (AHG) was positive. Screening of mother's serum for irregular antibodies showed anti-Jka antibody. AHG phase titers using tube technique were 1:64. Mother was found to be Jka antigen negative; father and neonate were found to be Jka antigenpositive. Antibody was observed to be of IgG type on Dithiothreitol treatment. Baby had total serum bilirubin of 20.5 mg/dl on day 3 of life and phototherapy was started. Exchange transfusion was not required in the baby. The present case emphasises the significance of minor blood group antigens other than Rh blood group system as a cause of HDFN. Although HDFN due to Jk antibodies is rare, however, the clinician must be aware of the occurrence of these antibodies as they can lead to severe HDFN and persistent anemia in the infant.

Segregation of blood inventory: A key driver for optimum blood stock management in a resource-poor setting.

BACKGROUND: Blood inventory management entails maintaining a delicate balance between guaranteeing blood availability and minimizing wastage. The study was conducted to identify and analyze various factors of wastage which can provide insight to ideal inventory management, thus help in formulating policies and improve efficiency of blood transfusion services. MATERIALS AND METHODS: The study was conducted in a tertiary care hospital. To determine various causes of wastage, a retrospective analysis was done over 6 months and preventive strategies adopted. Issuable stock index (ISI) and wastage as percentage of issue (WAPI) were used to compare the effect on blood inventory before and after adoption of strategies. The average number of times each ABO group and Rh type was crossmatched before final transfusion was calculated and compared for randomly selected units over the first 6 months of 2012 and 2013. RESULTS: Outdating was found to be the largest cause, and decrease in discarding rate was observed after adoption of strategies. Mean ISI for different study periods was comparable. However, significant decrease (P = 0.015) was observed for WAPI and WAPI with respect to outdating. Significant decrease in average number of times a unit was crossmatched before final transfusion for all positive blood groups and O-negative blood group was observed over corresponding first 6 months of 2012 and 2013. CONCLUSION: Division of inventory into two parts, enlistment of soon to outdate blood components, and reduction of holding of blood units to minimum period for elective surgery patients are simple measures which can minimize wastage.

Antibody screening in multitransfused patients: a prerequisite before each transfusion.

Life-long red blood cell (RBC) transfusions remain the main treatment for severe thalassemia. We hereby report a case of anti S and anti Lu(a) in a ß-thalassemia major patient detected incidentally on antibody screening. The patient was a known case of ß-thalassemia major and was on regular blood transfusion every 3 weeks from the institute from the age of 6 months. Subsequently, on one occasion, patient's crossmatch was compatible despite positive antibody screen using microcolumn gel technique. Autocontrol and direct antiglobulin test were negative on microcolumn gel. Anti S and anti Lu(a) antibodies were identified. Blood unit found compatible was negative for S and Lu(a) antigens. Antibody titers were 1:1 for both anti S and anti Lu(a) in AHG phase using tube technique and antibodies were of IgG type. Blood unit was transfused uneventfully to the patient. Donors were traced back (last three donations) and called for repeat blood sample testing for S and Lu(a) antigen. Two out of three donors were found to be S antigen positive and one out of these two was Lu(a) antigen positive. Anti S and anti Lu(a) antibodies were again identified on patient's subsequent visit for transfusion. The present case re-emphasize the importance of antibody screening at each visit in earlier detection of antibodies in multi transfused patients. Encouraging patients to receive transfusion from one center and dedicating donors could reduce alloimmunization rate but larger studies are required.

Assessment of Impact of Training in Improving Knowledge of Blood Transfusion among Clinicians.

BACKGROUND: Blood is a precious resource that needs to be prescribed, handled, stored and transfused as per guidelines to ensure recipient safety. The present study aims to assess the basic knowledge of clinicians pertaining to safe transfusion practice, impart relevant training, and assess the impact of such training programs. METHODS: A total of 25 fresh bachelor of medicine and bachelor of surgery graduates were enrolled for the study. The participants were given a pre-assessment questionnaire related to the entire transfusion chain followed by interactive training of the participants and post-training re-assessment. RESULTS: The mean score in the pre-training assessment was 51% while in the post-training assessment the mean score was 85.4%; the difference was statistically significant. There were significant differences in knowledge pertaining to storage temperature, shelf life of red cells and platelets, alternate group choice for fresh frozen plasma, and documentation of transfusion reaction. The participants had inadequate knowledge pertaining to cross-match procedure and management of transfusion reactions. CONCLUSION: The study assessed the knowledge and awareness of clinicians regarding blood transfusion practice. Mandatory training and inclusion of transfusion medicine as a subject at undergraduate level can help in improving transfusion practice and ensuring recipient safety.