Autoimmune anti-D in an RhD-positive young infant: Learning from a rare case.

BACKGROUND AND OBJECTIVES: Anti-D is usually immune in nature and is formed in individuals lacking D antigen or having variants/altered D phenotypes. In the Indian population, 93.8% are RhD positive, and R1 R1 is the commonest Rh phenotype. Here we report a rare and interesting case of autoimmune anti-D in an RhD-positive 3-month-old infant leading to warm autoimmune haemolytic anaemia. STUDY DESIGN AND METHODS: Auto-anti-D was detected serologically by immunohaematological techniques such as direct antiglobulin test, antibody detection and identification, dithiothreitol, enzyme treatment, antibody titration and elution. Molecular studies were performed to rule out genetic variants of RhD. RESULTS: Anti-D was confirmed in eluate and blood group post elution was B RhD positive. On genotyping using the Indian-specific RHD genotyping assay, the sample was found to be negative for the RHD*01W.150 (most common RhD variant in Indians) but positive for RHD exon 5 and RHD exon 10 along with glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The sample was further sequenced for RHD exons 1-10 by Sanger sequencing and found to be a wild type, thus, ruling out the presence of an RhD variant. CONCLUSION: This case is of interest because of the rare occurrence of autoimmune anti-D in an RhD-positive patient of such a young age (3 months). To the best of our knowledge, only two case reports have been published on autoimmune anti-D in infancy (in 1961 and 1964).

Challenges in the transfusion management of a 15-month-old pediatric patient with Bombay blood group phenotype.

Bombay blood group phenotype is often mistyped as O group which can lead to hemolytic transfusion reactions. There are a very few case reports of Bombay blood group phenotype in pediatric age group. Herein, we report an interesting case of Bombay blood group phenotype in a fifteen-month-old pediatric patient who presented with features of raised intracranial pressure and required an emergency surgery. The Bombay blood group was detected on detailed immunohematology work up which was further confirmed by molecular genotyping. The challenges faced in developing countries for transfusion management of such a case have been discussed.

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.

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.

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.

An ABO blood grouping discrepancy: Probable B(A) phenotype.

In B(A) phenotype, an autosomal dominant phenotype, there is a weak A expression on group B RBCs. We herein report a case of a probable B(A) phenotype in a first time 20-year old male donor. The cell and serum grouping were done using tube technique and also with blood grouping gel card (Diaclone, ABD cards for donors, BioRad, Switzerland). The antisera used were commercial monoclonal IgM type. To check for the weak subgroup of A, cold adsorption and heat elution was performed. The cell grouping was AweakB RhD positive while the serum grouping was B. There was no agglutination with O cells and the autologous control was also negative. It was a group II ABO discrepancy with or without group IV discrepancy. Results for both the eluate and last wash were negative. Hence, the possibility of weak subgroup of A was unlikely. Blood grouping gel card also showed a negative reaction in the anti-A column. One lot of anti-A was showing 'weak +' agglutination while the other lot was showing 'negative' reaction with the donor RBCs by tube technique. There was no agglutination observed with anti-A1 lectin. Our case highlights the serological characteristics of a B(A) phenotype. This case emphasizes the vital role of cell and serum grouping in detecting such discrepancies especially in donors which can lead to mislabeling of the blood unit and may be a potential risk for the transfusion recipient if not resolved appropriately.

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.

High Titer Antibodies Demonstrating Prozone in Two Repeat Blood Donors.

Introduction: - Prozone phenomenon due to excess unbound antibodies may sometimes lead to failure of detection of ABO incompatibility. . The case series describes immunohematology work up of blood group discrepancies of two blood donors. Materials and Methods: - Blood grouping was performed by a fully automated immune hematology analyzer (FAIHA Diagast, Qwalys 3, France) based on the principle of erythrocyte magnetized technology. Further immunohematology workup was done by tube technique (at different temperatures and phases) and column agglutination technique (CAT). Antibody titration was done by tube technique at saline and AHG (anti human globulin) phase. Results: - Type I blood group discrepancy was detected on initial blood grouping done by an automated analyzer. The discrepancy was resolved by repeat blood grouping by tube technique with a remarkable finding of hemolysis in reverse grouping. The lysis was attributed to high titer antibodies (anti-B titer of 512) with demonstration of prozone phenomenon. However, there was no discrepancy between cell and serum grouping by column agglutination technique (CAT). Conclusion: - Tube technique is the gold standard method for blood grouping and detects blood group discrepancies optimally. Hemolysis which is taken as a positive result, can be best appreciated by tube technique.

Autoimmune hemolytic anemia due to auto anti-N in a patient with sepsis.

We describe a case of autoimmune hemolytic anemia (AIHA) due to anti-N in a young male patient with cellulitis. There have been several reports of anti-N in N positive individuals. But in all these reports, auto anti-N was mostly associated with underlying immunological conditions. We report here a case of auto anti-N in a patient of bacterial sepsis without any underlying immune disorder.

Just not cosmesis! Role of low-density lipoprotein apheresis in familial hypercholesterolemia: Experience at a newly developed tertiary care institution in Northern India.

Familial hypercholesterolemia (FH) is characterized by an increase in plasma low-density lipoprotein-cholesterol (LDL-C) levels. It presents with tendon/skin xanthomas and premature atherosclerotic cardiovascular disease. The most available treatment options for FH are lipid-lowering medications such as statins, lifestyle modification, and LDL apheresis. As per American Society for Apheresis guidelines 2019, the treatment of FH using LDL apheresis falls under Category I. Here, we are reporting an interesting case of a young patient who presented with chief complaints of progressively increasing yellowish lesions around eyes, neck, hands, and legs. She was thoroughly investigated and was diagnosed provisionally as a case of Type 2 FH. Her total serum cholesterol and LDL-C were 717.2 mg/dl and 690.6 mg/dl, respectively, at presentation. One cycle of LDL apheresis was planned for her. We found immediate post-procedural reduction of 55.8% and 55.3% for total serum and LDL cholesterol levels respectively while 70.58% and 77.41% reduction in the levels from the day of presentation to the hospital.