A study on diagnostic performance of different immunohematological diagnostic tests in assessing the prevalence of ABO Hemolytic Disease of Newborn in the antenatal O group mothers and their neonatal outcome in a tertiary care hospital in Northern India.

BACKGROUND: Hemolytic disease of the newborn (HDN) results in the decreased lifespan of the red cells. HDN related to ABO incompatibility is mostly unnoticed because routine screening is not being done. This study was done to assess the prevalence of ABO-HDN and to compare different immunohematological tests. Methods-In this study 213 O group mothers and the 122 ABO-incompatible newborns born to them were included. Quantifying the maternal IgG anti-A/anti-B antibody titer was done by Conventional Tube Technique (CTT) using Dithiothreitol (DTT) pretreated maternal serum. Hemolysin test was performed on the mothers having titer > 256. These cases were followed up and, after delivery, were monitored for ABO HDN, along with direct antiglobulin testing and elution studies. The prevalence of ABO-HDN was calculated, and the different diagnostic parameters of the tests were calculated. Results- The prevalence of ABO-HDN in our population was estimated to be 1.7%, 6.1% & 10.6% in our population, O group mothers, and O group mothers with ABOincompatible newborns, respectively. Maternal titer≥ 512 strongly correlated with ABOHDN. DAT positivity is a good predictor of ABO-HDN, especially using sensitive techniques. Maternal IgG titers have the highest sensitivity & Negative Predictive Value, while DAT has the highest specificity & Positive Predictive Value. Conclusion - Maternal ABO antibody titration may be advocated in the centers to identify high-risk groups. It can advocate institutional delivery and dedicated follow-up of newborns with ABO-HDN. Blood grouping & DAT may be performed in all newborns born to O blood group to identify high-risk cases.

Off with the blues: Therapeutic plasma exchange in a case of copper sulphate poisoning.

CuSO4 (Copper sulphate) poisoning though rare, is associated with high mortality. It involves multiple organ systems and if not dealt with promptly can lead to death. Supportive care and chelation therapy along with TPE (therapeutic plasma exchange), whole blood exchange or red cell exchange can be employed in management. We report such a case where swift clinical improvement was seen after TPE.

Irregular erythrocyte antibodies among antenatal women and their neonatal outcome at a tertiary care hospital in Northern India.

BACKGROUND: Red blood cell alloimmunisation during the pregnancy is a significant cause for neonatal mortality and morbidity. This study was planned to determine the prevalence and specificity of irregular erythrocyte antibodies in antenatal mothers and their neonatal outcome. METHODS: In this observational study, blood grouping and red cell antibody screening of mothers were performed at first visit and after 28 weeks of gestation and positive cases were identified and followed up monthly till delivery by repeating antibody titre and middle cerebral artery-peak systolic velocity. After delivery of alloimmunised mothers, cord blood haemoglobin, bilirubin and direct antiglobulin tests (DAT) were analysed and further outcome of neonate was recorded. RESULTS: Among 652 registered antenatal cases, 18 multigravida women were found to be alloimmunised, accounting to prevalence of 2.8%. Most common alloantibody identified was anti D (>70%) followed by anti-Lea, anti-C, anti-Leb, anti-E and anti-Jka. Only 47.7% Rh D negative women received anti-D prophylaxis during previous pregnancies or whenever indicated. DAT was positive in 56.2% of neonates. Among nine DAT positive neonates, two early neonatal deaths due to severe anaemia were observed following birth resuscitation. Four antenatal mothers required intrauterine transfusion in view of fetal anaemia while three neonates received double volume exchange transfusion and top up transfusions after birth. CONCLUSIONS: This study emphasises importance of red cell antibody screening for all multigravida antenatal women at registration of pregnancy and additionally at 28 weeks or later in high-risk cases irrespective of RhD status.

D or anti-D!!! Unblocking the dilemma of blocking-D phenomenon using acid elution.

Maternal IgG antibodies directed against fetal red cells can cause hemolytic disease in fetus and newborn manifesting as anemia and jaundice. Sometimes, these antibodies are so strong that they encapsulate the antigens on neonatal red blood cells and result in erroneous laboratory findings when tested. A requisition for double volume exchange transfusion was received for a term,3.1 kg female baby with neonatal jaundice at day 2 of life, born to a multiparous woman. The neonate was typed as AB RhD negative and the mother as A Rh D negative. The maternal sample tested positive for Indirect antiglobulin testing showing presence of Anti-D with IgG titer of 128. The direct antiglobulin testing for baby was strongly (4 +) positive. The strong DAT result with negative RhD typing for the neonate indicated towards the Blocking-D phenomenon. We attempted to resolve the Blocked-D case using acid elution, which revealed the presence of D antigen on the eluted neonate's red cells. The report emphasizes the importance of appropriate blood typing for neonates to provide prompt adequate care as a team by the departments of Neonatology and Transfusion Medicine.

A prospective interventional study to assess the impact of a 'structured compact training' on knowledge and skills of safe blood transfusion practices among nurses working in a tertiary care institute.

INTRODUCTION: There is scarce information on the baseline knowledge and practices of nursing officers in relation to administration of blood components. We set out to evaluate the influence of training on their knowledge and skills through Kirkpatrick's levels of Training Evaluation. MATERIALS AND METHODS: This interventional cross sectional study of 7 months duration conducted in a tertiary care teaching institute involved 200 nursing officers. Hundred were assigned to study/intervention group and 100 were assigned to control/ comparison group by systematic random sampling. Knowledge was tested in different domains-blood components, pre-transfusion checks, transfusion process, post-transfusion process and blood administration practice. RESULTS: The baseline knowledge scores of intervention and control group were similar-15.16 ± 4.11 and 15.02 ± 4.75 (p = 0.831). Post-intervention (phase I) after 1 month, the scores improved significantly for domain A, B, C, D and E to 4.3 ± 2.21 (p = 0.0001), 3.46 ± 2.15 (p = 0.0001), 7.02 ± 3.55 (p = 0.0001), 2.51 ± 1.46 (p = 0.0012), and 5.86 ± 3.61 (p = 0.0018) respectively. In phase II, after 3 months of training, and the scores were significantly better from baseline for all domains except E. For domain A, B, C, D and E, scores were 3.82 ± 2.46 (p = 0.0001), 3.53 ± 1.98 (p = 0.0001), 7.38 ± 3.87 (p = 0.0001), 2.48 ± 1.55 (p = 0.0035), and 5.86 ± 3.61 (p = 0.95) respectively. CONCLUSIONS: Our study showed that baseline scores were low in the nursing officers. No significant difference was found in baseline scores in subject and control population. However, post-intervention, a significant improvement in scores was observed in the study group across all domains.

COVID 19 infection associated with thrombotic thrombocytopenic purpura.

Thrombotic thrombocytopenic purpura (TTP) which can cause significant mortality is a thrombotic microangiopathy due to deficiency of VWF cleaving protease ADAMTS13 and as per medical literature there are examples that TTP can be caused by COVID 19 infection. A 35 years old female after admission with right sided weakness and slurring of speech was found to be COVID positive and diagnosed as a case of TTP. Patient had absent ADAMTS13 level on day 1. Treatment was started with therapeutic plasma exchange (TPE) later injection Vincristine and Rituximab was given after 4th TPE as it was suspected as refractory case. Finally patient received 16 TPE procedures with cryo poor plasma as exchange fluid and gradually her platelet count started to maintain normal and she was discharged. Specific management and such association of this type of cases need to be studied more judiciously.

Irregular erythrocyte antibodies among antenatal women and their neonatal outcome at a tertiary care hospital in Northern India.

BACKGROUND: Red blood cell alloimmunisation during the pregnancy is a significant cause for neonatal mortality and morbidity. This study was planned to determine the prevalence and specificity of irregular erythrocyte antibodies in antenatal mothers and their neonatal outcome. METHODS: In this observational study, blood grouping and red cell antibody screening of mothers were performed at first visit and after 28 weeks of gestation and positive cases were identified and followed up monthly till delivery by repeating antibody titre and middle cerebral artery-peak systolic velocity. After delivery of alloimmunised mothers, cord blood haemoglobin, bilirubin and direct antiglobulin tests (DAT) were analysed and further outcome of neonate was recorded. RESULTS: Among 652 registered antenatal cases, 18 multigravida women were found to be alloimmunised, accounting to prevalence of 2.8%. Most common alloantibody identified was anti D (>70%) followed by anti-Lea, anti-C, anti-Leb, anti-E and anti-Jka. Only 47.7% Rh D negative women received anti-D prophylaxis during previous pregnancies or whenever indicated. DAT was positive in 56.2% of neonates. Among nine DAT positive neonates, two early neonatal deaths due to severe anaemia were observed following birth resuscitation. Four antenatal mothers required intrauterine transfusion in view of fetal anaemia while three neonates received double volume exchange transfusion and top up transfusions after birth. CONCLUSIONS: This study emphasises importance of red cell antibody screening for all multigravida antenatal women at registration of pregnancy and additionally at 28 weeks or later in high-risk cases irrespective of RhD status.

Severe hemolytic disease of the fetus and newborn due to anti-E and anti-Jka.

CONCLUSIONS: Red blood cell alloimmunization to antigens other than D, such as C, c, E, e, and antigens in the Kell, MNS, and Duffy blood group systems, has emerged as an important cause of hemolytic disease of the fetus and newborn (HDFN). Antibody screening for these antibodies is not routinely practiced for all antenatal patients in developing countries, mainly because of financial constraints. Here we report a rare case of HDFN due to dual antibodies to Rh and Kidd blood group system antigens: anti-E and anti-Jka. This case report highlights the importance of routine and regular antenatal screening of all pregnant women for proper monitoring and follow-up.

Routine indirect antiglobulin testing of blood donors-a further step toward blood safety: an experience from a tertiary care center in northern India.

CONCLUSIONS: This scientific article emphasizes the importance of a policy for antibody screening of all blood donors as a step to further improve blood safety. We also report the incidence of red blood cell (RBC) alloimmunization in healthy blood donors obtained using a cross-sectional prospective study from September 2017 to January 2019 in the Department of Transfusion Medicine of a tertiary care referral and teaching institute in northern India. The indirect antiglobulin test (IAT) for unexpected RBC antibodies was performed by the conventional tube test with pooled group O RBCs on all donor units irrespective of their D status. Samples with positive IATs were sent to the Immune Hematology Reference Laboratory for further immunohematolo-gy workup, maintaining predefined optimal storage and transport conditions. Of the 10,390 donors studied, 9959 were males and 431 were females. The incidence of unexpected antibodies (antibodies other than those of the ABO blood system) among the blood donors was found to be 0.18 percent (19 of 10,390 with 25 alloantibodies). Of the 19 alloimmunized donors, 16 (84.2%) were male (alloimmunization rate 0.16%, 16 of 9959) and 3 (15.8%) were female (alloimmunization rate 0.69%, 3 of 431) (p = 0.01; chi-square test). In our study, the most frequent alloantibodies identified were of the Lewis blood group system (17 of 25 [68%] in 14 of the 19 alloimmunized donors). The second most common allo-antibodies belonged to the Rh blood group system (4 of 25 [16%] in 3 of the 19 alloimmunized donors), followed by those of the MNS blood group system (3 of 25 [12%] in 2 of 19 alloimmunized donors). Anti-K was found in one donor (1 of 25 [4%]). Based on the results of the study, we recommend that a policy of routinely performing IATs on all donor units, irrespective of their D status, be adopted as an essential component of safe blood transfusion practices.

Clinically significant naturally occurring anti-N and anti-S in a blood donor: a rare finding.

CONCLUSIONS: To the Editors: Alloimmunization is triggered when an individual whose red blood cells (RBCs) are lacking particular antigens is exposed to these antigens through transfusion or pregnancy, causing the formation of immune antibodies. In addition to these exogenous exposures, underlying inflammatory or autoimmune conditions may lead to formation of unexpected antibodies. Individual factors also play a role, since some people are responders and others are non-responders. We report a case of naturally occurring alloanti-N and alloanti-S in a healthy D+ blood donor. Both antibodies were reactive over a wide thermal amplitude and hence were potentially clinically significant. This case highlights the importance of incorporating the indirect antiglobulin test (IAT) to test for unexpected RBC antibodies for all blood units as a routine protocol.

Hematological parameters in patients of cleft lip and cleft palate with special reference to eosinophil counts.

INTRODUCTION: Birth abnormalities like cleft lip and cleft palate account for about 1.4 per 1000 live births in India. These are seen to be associated with a high incidence of eosinophilia which delays the surgical management of these patients. AIMS: The aim of this paper is to study the hematological parameters in patients of cleft lip and cleft palate. PATIENTS AND METHODS: A total of 223 cases of cleft lip and cleft palate were taken up for the study. Hematological parameters including hemoglobin, total leukocyte count, differential leukocyte count, absolute eosinophil count, and red cell indices were studied. RESULTS: Anemia was found in 182/223 (81.63%) cases which was most commonly of microcytic hypochromic type. Eosinophilia was seen in 46/223 (20.60%) cases. CONCLUSIONS: Many cleft lip and cleft palate patients show high eosinophil counts. Absolute eosinophil count was found to be a better parameter for assessment of eosinophils.

Utility of COVID-19 Seropositive Plasma as Convalescent Plasma: An Immune and Neutralization Antibody Seroprevalence Analysis in Blood Donors for Future Potential Pandemic Readiness.

OBJECTIVES: To analyze the seroprevalence of SARS-CoV-2 IgG antibodies and neutralizing antibodies in blood donors during the second wave of the pandemic and to explore the utility of COVID-19 seropositive plasma as convalescent plasma. MATERIALS AND METHODS: In this study, 696 blood donors were tested for anti-SARS-CoV-2 IgG antibodies using a chemiluminescence assay. By blinding, 271 samples were chosen randomly for testing of neutralizing antibodies by enzyme-linked immunosorbent assay (ELISA) in duplicate among the 696 blood donors tested for anti-SARS-CoV-2 IgG antibodies, irrespective of the positivity or negativity of the result of the anti-SARS-CoV-2 IgG antibodies by chemiluminescence assay. IgG antibody levels were analyzed in signal-to-cutoff (S/Co), while neutralizing antibody levels were analyzed in percentage inhibition. RESULTS: The seroprevalence of IgG antibodies based on the S/Co for the positive results ≥ 1.00 was 82.75%, while the seroprevalence of neutralizing antibodies based on the percentage inhibition for the positive results ≥ 30% was 89.59%. Frontline workers (FLWs) and Covishield-vaccinated individuals showed higher levels of the anti-SARS-CoV-2 IgG antibodies regarding higher S/Co. In comparison, levels of neutralization antibodies regarding percentage inhibition were higher only in FLWs. Covishield-vaccinated donors elicited a statistically higher seroprevalence of anti-SARS-CoV-2 IgG antibodies compared to the Covaxin-vaccinated, while the seroprevalence of neutralizing antibodies was not statistically different among this group. There was a positive correlation (0.762) between anti-SARS-CoV-2 IgG antibodies and neutralizing antibodies, and almost all donors' of S/Co ≥ 9.5 had neutralizing antibodies. CONCLUSION: This study showed higher seroprevalence in the blood donor population compared to published seroprevalence in India's second wave of the pandemic. In the current study, 328 donors (47.12%) of the 696 screened donors were neither vaccinated nor had previous SARS-CoV-2 infection, but many had antibodies. The seroprevalence of neutralizing antibodies (96.42%) was higher than the seroprevalence of the anti-SARS-CoV-2 IgG antibodies (85.71%) in the donors who had previous infection of COVID-19. On the other hand, vaccinated donors showed similar immune responses for neutralizing antibodies and the anti-SARS-CoV-2 IgG antibodies. Higher IgG immune reactivity in S/Co showed a good correlation with neutralizing antibodies and can be used to screen whole blood donors for convalescent plasma donations.

Therapeutic Leukocytapheresis to relieve priapism due to leukostasis in CML patients: Our experience in a tertiary care centre.

Hyperleukocytosis in leukemic patients may cause tumour lysis syndrome, disseminated intravascular coagulopathy, and leukostasis, resulting in decreased tissue perfusion and increasing the risk of mortality. Since the myeloid blasts are larger than lymphoid blasts and are less deformable, complications of leukostasis are seen more frequently in myeloid leukemia. Priapism is a less common complication associated with leukostasis in leukaemia patients that should be treated as soon as possible to avoid ischemic injuries. Although chemotherapeutic drugs such as hydroxyurea and imatinib are used to treat hyperleukocytosis in CML patients, leukocytapheresis (LCP) can achieve rapid cytoreduction. Prophylactic LCP could not offer any advantage over aggressive chemotherapy, but therapeutic leukocyte depletion has a proven role in patients having symptomatic leukostasis due to high tumour burden. Three patients with ischaemic priapism were reported at our institute's emergency department, where detumescence could not be achieved by distal shunting or aspiration with phenylephrine instillation. The procedure of therapeutic LCP was performed in all three patients on an emergency basis, which resolved painful priapism by rapid cytoreduction.

Analysis of reasons of blood donor deferral at a tertiary care institute in India and its reflections on community health status.

INTRODUCTION: Safe blood donors form the backbone of safe blood transfusion services.[1] Donor eligibility policies are a critical layer of blood safety designed to ensure selection of healthy donors and to protect recipients from any harm. This study was planned to analyze the pattern of whole blood donor deferrals and its characteristics and reasons at a tertiary care institute in northern India, as the pattern varies according to epidemiology of diseases in different demographic areas. MATERIALS AND METHODS: It was a cross-sectional study of 2 years' duration from December 2015 to November 2017. The data of the potential donors who were deferred were recorded on a separate pro forma which included their demographic details, type of donation - voluntary donor and replacement donor; first time and repeat donor; type of deferrals (permanent and temporary); and the reasons of deferrals. RESULTS: A total of 3133 donors (voluntary - 1446 and replacement - 1687) donated and 597 donors were deferred (deferral rate - 16%) during this period. Majority of the deferrals, i.e., 525 (88%) were temporary, while 72 (12%) were permanent. The most common reason of temporary deferral was anemia. The most common reason of permanent deferrals was a medical history of jaundice. CONCLUSIONS: Our study results indicate that the blood donor deferral can have subtle variations based on regional aspects that should be considered when national policies are developed as pattern of deferral varies according to the epidemiology of diseases in different demographic areas.

Structured feedback: A teaching and learning tool for the postgraduate medical students in transfusion medicine.

CONTEXT: Structured Feedback is a learning and assessment tool designed to provide feedback to students and educators to adjust learning and teaching during the training. Lack of provision of structured feedback to postgraduate (PG) medical students prompted us to plan a study to introduce a structured feedback module into the existing monthly assessment schedules in the Department of Transfusion Medicine. AIM: This study aims to introduce a structured feedback module and evaluate its efficacy after incorporation into the existing monthly assessment schedules for the PG students in the Department of Transfusion Medicine. DESIGN AND SETTING: A quasi-experimental study was commenced after obtaining clearance from the Institutional Ethics Committee in the Department of Transfusion Medicine for the students pursuing postgraduation in Transfusion Medicine. METHODOLOGY: A peer-validated feedback module was designed and implemented for MD students by the core team faculty. The students underwent the structured feedback sessions after each monthly assessment for of 3 months. One on one, verbal feedback was conducted using Pendleton's method, for monthly online assessment for the learning that happened during study period. DATA COLLECTION AND STATISTICAL ANALYSIS: The data were collected from the open-ended and closed-ended questions using Google form-based Student/Faculty perception and students' pre-post self-efficacy questionnaires on 5-point Likert Scale and the quantitative data analysis was done using percentage of Likert scores, median values for each item for pre-and post-responses and comparison using nonparametric test - Wilcoxon signed-rank test. The qualitative data analysis was done using thematic analysis from the open ended questions. RESULTS: All (n = 9; 100%) the PG students strongly agreed and agreed (median score of 5 and 4) that the feedback they received made them aware of their learning gaps, enabled them in bridging those gaps and provided ample opportunity to interact with faculty. Both students and faculty agreed that the feedback session should be an ongoing and continuous process in the department. CONCLUSION: Both the students as well as faculty were satisfied with the implementation of the feedback module in the department. Students reported awareness about the learning gaps, identification of appropriate study resources, and ample opportunity to interact with faculty, after taking the feedback sessions. The faculty felt satisfied on the acquisition of new skill for delivering structured feedback to students.

Association of donor characteristics with coagulation factor levels in fresh frozen plasma.

BACKGROUND: Coagulation factors are essential to maintain normal hemostasis. Plasma for transfusion can be obtained from whole blood donation or plasma apheresis. Plasma obtained from whole blood donation is termed as fresh frozen plasma (FFP). The quality of FFP can be influenced by several factors including donor variables (such as age, gender, diet, genetic profile), environmental factors, collection methods, processing methods, storage temperature, etc. This study was done to assess the association of donor characteristics such as donor age, blood group, and smoking with coagulation factor levels in FFP units. MATERIALS AND METHODS: The screening of donors for collection of whole blood units was done as per the national guidelines. A total of 144 FFP units were assessed for coagulation factors. The FFP units were tested for prothrombin time (PT), activated partial thromboplastin time, fibrinogen, coagulation factor VIII, and coagulation factor IX (CF IX) on coagulation analyzer. RESULTS: A total of 144 FFP units were tested for coagulation parameters. The value of PT was highest in units prepared from donors in more than 45 years of age group. The value of CF IX was significantly lower in O blood group as compared to non-O blood group. The value of fibrinogen was significantly higher in smokers as compared to nonsmokers. CONCLUSION: The findings of the present study further add evidence to the fact that donor factors such as age, blood group, and smoking have an impact on coagulation factor levels in FFP units.

Convalescent Plasma the Experience and Journey from Blockbuster to Incognito: A Single Centre Experience.

BACKGROUND: Convalescent plasma has been used to provide passive immunotherapy to patients with COVID-19 with a high level of safety. Very few efficacy studies were available, and due to COVID being a relatively new disease, its exact therapeutic role was unclear. This observational study on the impact of COVID convalescent plasma (CCP) on clinical outcomes attempts to evaluate the effectiveness of convalescent COVID-19 plasma therapy in the treatment of COVID-19 patients at the tertiary care center in the Uttarakhand state of India. METHODS: CCP was collected by plasmapheresis/whole blood from willing COVIDrecovered donors who underwent pre-donation testing including ABO and RhD grouping, mandatory blood screening tests for HIV, HBV, HCV, syphilis and Malaria, Haemoglobin estimation and COVID IgG assay. Hospitalized patients with severe COVID-19 pneumonia who received these CCP units were followed up and the outcome (Recovery/death) was observed. RESULTS: A total of 63 patients who received CCP were included in the study. Out of the total, 13 (20.7%) were females and 50 (79.3%) were males and their ages ranged from 24 to 80 years with a median age of 53 years. The period between the start of symptoms and hospitalization ranged from 1 to 14 days with an average duration of 4.7 days. Symptoms on presentation included Fever 53/63 (84.1%), Tachypnoea 60/63 (95.2%) and Cough 42/63 (66.7%). Among these patients, 22/63 (34.9%) were on non-invasive ventilation (NIV), 6/63 (9.5%) on non-rebreather mask (NRBM) and 32/63 (50.8%) were on Ventilator support. The infused convalescent plasma had a Mean IgG value of 57.3 AU with a range of (10-142 AU). A total of 37 (58.7%) patients were lost to COVID-19 infection and 26 (41.3%) were discharged from the hospital in a healthy state. CONCLUSION: The use of convalescent plasma in addition to standard treatment in our study on patients with severe pneumonia due to COVID-19 did not demonstrate reduced mortality of COVID-19 patients amidst numerous variables. The results showed that the use of convalescent plasma as a treatment option in the present conditions needs a serious re-evaluation. Studies on a strictly defined recipient group and transfusion of CCP units, with adequate antibody titer and/or neutralization activity, must be analyzed for future works.