Effect of phenotypes of Duffy blood group on chemokine storage and chemokine scavenging function of erythrocytes / 中国输血杂志

【Objective】 To investigate the effect of phenotypes of Duffy blood group on chemokine storage and chemokine scavenging function of erythrocytes. 【Methods】 Twenty-four erythrocyte samples were collected and tested Duffy blood phenotype using the anti-human globulin method, and erythrocyte CCL2, CCL5, CXCL8, and CCL11 content and their chemokine scavenging function using ELISA. The expression of Duffy antigens on erythrocytes was detected using a flow analyzer. 【Results】 The difference in CCL2 content(41.1±14.7 pg/mL vs 63.1±20.8 pg/mL)of erythrocyte lysate between Fy(a+b-) and Fy(a+b+) phenotype was statistically significant (P0.05).The difference in the scavenging function of CCL2(1471±202.1 pg/mL vs 1860±267.5 pg/mL)and CCL5 (848.5±461.7 pg/mL vs 1797±546.1pg/mL) between Fy(a+b-) and Fy(a+b+) phenotype were statistically significant (P0.05).The difference in Duffy antigen expression (mean fluorescent intensity:105.3±20.45 vs 111.9±18.30)on erythrocytes between Fy(a+b-) and Fy(a+b+) phenotype was not statistically significant (P>0.05). 【Conclusion】 The Fy(a+b+) and Fy(a+b-) phenotypes of the Duffy blood group can affect the chemokine storage and scavenging function of erythrocytes. Fy(a+b+) phenotypes are able to store more chemokines and have a stronger chemokine scavenging function than Fy(a+b-) phenotypes.

Association between FY* 02N. 01 and the severity of COVID-19: initial observations

ABSTRACT Introduction The pro-inflammatory immune response underlies severe cases of COVID-19. Antigens of the Duffy blood group systems are receptors for pro-inflammation chemokines. The ACKR1 c.-67T>C gene variation silences the expression of Duffy antigens on erythrocytes and individuals presenting this variant in homozygosity have impaired inflammatory response control. Our aim was to evaluate the association between the ACKR1 c.-67T>C and the severity of COVID-19. Methods This was a retrospective single-center case-control study, enrolling 164 participants who were divided into four groups: 1) Death: COVID-19 patients who died during hospitalization; 2) Hospital Discharge: COVID-19 patients who were discharged for home after hospitalizations; 3) Convalescent Plasma Donors: COVID-19 patients who were not hospitalized, and; 4) Controls: patients with diagnosis other than COVID-19. Patients were genotyped for the ACKR1 c.-67T>C (FY*02 N.01 allele) and the frequency of individuals presenting the altered allele was compared between the groups. Results The groups significantly differed in terms of the percentage of patients presenting at least one FY*02N.01 allele: 36.8% (Death group), 37% (Hospital Discharge group), 16.1% (Convalescent Plasma group) and 16.2% (Control group) (p= 0.027). The self-declared race (p < 0.001) and the occurrence of in hospital death (p= 0.058) were independently associated with the presence of the FY*02N.01 allele. Hypertension (p < 0.001), age (p < 0.001) and the presence of at least one FY*02N.01 allele (p= 0.009) were independently associated with the need for hospitalization. Conclusion There is a suggestive association between the presence of the FY*02N.01 and the severity of COVID-19. This may be a mechanism underlying the worse prognosis for Afro-descendants infected with SARS-CoV-2.

Serological and Molecular Study of the Duffy Blood Group among Malarial Endemic Region Residents in Brazil

ABSTRACT Background: The atypical chemokine receptor 1 (ACKR1) gene encodes the Duffy blood group antigens in two allelic forms: FY*A (FY*01) and FY*B (FY*02), which define the Fy(a+b-), Fy(a-b+), and Fy(a+b+) phenotypes. FY*BES (FY*02N.01) is a single T to C substitution at nucleotide -67 that prevents the FY*B from being expressed in red blood cells (RBCs). Methods: We evaluated 250 residents from a Brazilian malarial endemic region (RsMR). All individuals were phenotyped for Fya and Fyb antigens and genotyped for FY*A, FY*B, FY*B SE , and FY*B weak alleles. Results: Among the 250 individuals, 209 (83.6%) reported previous malaria infection, and 41 (16.4%) did not. The Fy(a+b+) phenotype was present in 97/250 (38.8%), while the Fy(a-b-) was present in 7/250 (2.8%). The FY*A/FY*B was found in 130/250 (52%) and the FY*A/FY*A in 45/250 (18%). The c.1-67>TC was present, in homozygosity, in 11/250 (4.4%). Among 34 individuals with the Fy(a+b-) and FYA*/FYB* mutations, 4/34 (11.8%) had homozygosity for the c.1-67T>C. One individual presented the Fy(a+b-), FY*A/FY*B, and c.1-67T>C in homozygosis, whereas the other presented the Fy(a+b-), FY*A/FY*A, and c.1-67T>C in heterozygosis. Conclusions: We reported a low prevalence of the Fy(a-b-) in persons who had previously been infected with Plasmodium vivax (67.5%). We observed that 102/141 (72.3%) individuals expressing the Fyb antigen had a P. vivax infection, indicating the importance of the Fyb antigen, silenced by a c.1-67T>C mutation in homozygosis, in preventing the P. vivax infection. We showed that the c.1-67T>C mutation in the FY*A did not silence the FY*A expression on RBCs.

Genetic diversity of Duffy binding protein 2 region II of Plasmodium cynomolgi from wild macaques in Peninsular Malaysia

@#Recent reports of natural human infection by Plasmodium cynomolgi indicate the increased risk of zoonotic transmission by this simian parasite. The P. cynomolgi Duffy binding protein 2 (PcDBP2) has a potential role in the invasion pathway of host erythrocytes, and it is a possible vaccine candidate against cynomolgi malaria. This study investigates the genetic diversity, haplotypes, and natural selection of PcDBP2 region II from isolates collected from wild macaques in Peninsular Malaysia. Blood samples from 50 P. cynomolgi-infected wild macaques were used in the study. Genomic DNA extracted from the blood samples was used as template for PCR amplification of the PcDBP2 region II. The amplicons were cloned into a plasmid vector and sequenced. MEGA X and DnaSP ver.6.12.03 programmes were used to analyse the DNA sequences. A genealogical relationship of PcDBP2 region II were determined using haplotype network tree on NETWORK ver.10.2. Result showed high genetic diversity (ð = 0.017 ± 0.002; Hd = 1.000 ± 0.001) of the PcDBP2 region II. The Z-test indicates a purifying selection, with population expansion as shown in Tajima’s D analysis. A total of 146 haplotypes of PcDBP2 region II were observed. Phylogenetic tree analysis showed that these haplotypes were grouped into three allelic types (136 for Strain B type, 9 for Berok type, and 1 recombinant type). In the haplotype network, PcDBP2 region II revealed no geographical groupings but was divided into two distinct clusters.

Tipificación del antígeno Duffy como método indirecto para identificar individuos asintomáticos de malaria / Duffy antigen typing as an indirect method to identify asymptomatic malaria individuals

En países considerados endémicos de malaria a pesar de las estrategias realizadas anualmente para el control y erradicación de esta enfermedad existen brotes aislados de malaria debido probablemente a portadores asintomáticos, presencia de individuos con fenotipo Duffy negativo o movimientos poblacionales. El sector "50 casas" de la provincia de Esmeraldas-Ecuador reúne todas estas características por lo que el objetivo de esta investigación fue determinar la presencia del fenotipo Fy(a-b-) y su relación con el Plasmodium vivax. Se realizó una encuesta a cada miembro de las familias participantes y luego de aceptado y firmado el consentimiento informado se tomaron muestras sanguíneas para la realización de pruebas serológicas en búsqueda de anticuerpos y antígenos de P. vivax, fenotipificación del sistema Duffy y pruebas de PCR en tiempo real para la determinación de ADN de Plasmodium vivax y P. falciparum. Los resultados demostraron que existe una asociación estadísticamente significativa entre el fenotipo Duffy y malaria por P. vivax (p<0,05). Ante esta situación los organismos de control deberán proponer nuevas estrategias para la detección de portadores asintomáticos y medidas preventivas para evitar nuevos brotes de malaria, así como determinar si P. vivax ha encontrado un nuevo mecanismo de invasión en individuos portadores del fenotipo Fy(a-b-).

Despite the strategies carried out annually for the control and eradication in countries considered endemic for malaria, there are isolated outbreaks probably due to asymptomatic carriers. These involve the presence of individuals with Duffy negative phenotype or population movements. The neighbourhood "50 casas" of the province of Esmeraldas-Ecuador has all these characteristics, so the purpose of this research was to determine the presence of the Fy(a-b-) phenotype and its relationship with the Plasmodium vivax. A survey was carried out into each member of the participating families and after accepting and signing the informed consent, blood samples were taken to perform serological tests in search of antibodies and antigens of P. vivax. The phenotyping of Duffy system and real-time PCR tests for DNA determination of Plasmodium vivax and falciparum was realized. The results showed that there is a statistically significant association between the Duffy phenotype and P. vivax malaria (p<0.05). In view of this situation, the control agencies should propose new strategies for the detection of asymptomatic carriers and preventive measures to hinder new outbreaks of malaria and determine if P. vivax has found a new invasion mechanism in individuals with the Fy(a-b-) phenotype.

Nos países considerados endêmicos para malária, apesar das estratégias realizadas anualmente para o controle e erradicação desta doença existem surtos isolados de malária, resultantes provavelmente de portadores assintomáticos, presença de indivíduos com fenótipo Duffy negativo ou movimentos populacionais. O setor "50 casas" na província de Esmeraldas-Equador reúne todas essas características. Portanto o objetivo desta pesquisa foi determinar a presença de fenótipo Fy(a-b-) e sua relação com Plasmodium vivax. Foi realizada uma enquete para cada membro das famílias participantes e após a aceitação e assinatura do Consentimento informado, foram coletadas amostras de sangue para a realização de testes sorológicos em busca de anticorpos e antígenos do P. vivax. Além de testes de fenotipagem do sistema Duffy e PCR em tempo real para determinação de DNA de Plasmodium vivax e P. falciparum. Os resultados mostraram que existe uma associação estatisticamente significante entre o fenótipo Duffy e a malária por P. vivax (p<0,05). Perante está situação os órgãos de fiscalização terão que propor novas estratégias para a detecção de portadores assintomáticos e medidas preventivas para evitar novos surtos de medidas de malária bem como determinar se P. vivax tem encontrado um novo mecanismo de invasão em indivíduos portadores do fenótipo Fy(a-b-).

Reacción hemolítica transfusional tardía en un paciente con anemia de células falciformes: reporte de un caso / Delayed hemolytic reaction to transfusion in sickle cell anemia: report of one case

Sickle cell anemia was a rare disease in Chile, especially in adults, however the recent immigration wave from Haiti is changing this scenario. We report a 29 year old black female from Haiti with a non-disclosed history of sickle cell anemia. She was transfused with two units of red blood cells, found unconscious and with jaundice five days later and admitted to the hospital. On admission she had a hemoglobin of 3.3 g/dL, a total bilirubin of 5.08 mg/dL, a LDH of 1,306 Ui/L. She was transfused again, worsening her condition. An alloimmunization and delayed hemolytic reaction was suspected. A direct Coombs test was positive. She was treated with steroids and her serum hemoglobin rose progressively.

Clinical analysis of seven cases of rare hemolytic disease of the newborn / 中华儿科杂志

Objective@#To summarize the clinical features of 7 rare cases of hemolytic disease of newborn (HDN), and to improve the understanding of rare HDN.@*Methods@#Data of clinical information, laboratory findings, treatments and outcomes were collected and analyzed for four cases with HDN due to anti-M, two cases due to anti-Kidd, and one case due to anti-Duffy. All of them were admitted to the Department of Neonatology, Beijing Children's Hospital Affiliated to Capital Medial University from July 2007 to June 2017.@*Results@#Among the four MN hemolytic babies, two were males and two were females. Jaundice was found in three cases. Two cases had hyperbilirubinemia, one of them had severe hyperbilirubinemia. All the four cases developed anemia, including severe anemia in three cases. Two cases of Kidd hemolytic disease and 1 case of Duffy hemolytic disease had jaundice and anemia, but did not reach the level of severe hyperbilirubinemia and severe anemia. MN hemolytic disease babies got negative results in direct antiglobulin test, whereas the Kidd and Duffy hemolytic disease babies had positive findings in direct antiglobulin test. None of the babies had blood transfusion, and they were discharged from the hospital.@*Conclusions@#Without maternal and fetal blood group incompatibility (ABO or Rh blood-group system), for early onset of jaundice, severe jaundice or anemia, antiglobulin test to mother and child earlier should be administered, and MN, Kidd, Duffy and other rare hemolytic disease of the newborn should be pay attention to.

Case report and literature review of neonatal hemolysis induced by anti-E and anti-Ec combined with anti-Fyb / 天津医药

Hemolytic diseases of newborn (HDN) can cause miscarriage, premature birth, fetal edema, fetal intrauterine anemia and even fetal death in early pregnancy. Neonatus with HDN can have jaundice, anemia, hepatosplenomegaly, edema and nuclear jaundice sequelae. This article reviewed the diagnosis and treatment of two patients with HDN caused by anti-E and anti-Ec combined with anti-Fyb, and reviewed the relevant literature on the epidemilogy, the diagnosis and treatment of HDN in order to improve the understanding of the disease.

Identification of Anti-Gerbich Antibody in an Emirati Marrow Hematopoietic Progenitor Cell Donor with Fy(a−b−) Phenotype

In this study, we report a case of anti-Gerbich (Ge) alloantibody to a high-prevalence Ge antigen in a donor with Fy(a−b−) phenotype. The alloantibody was detected in an Emirati boy who was admitted to a Korean tertiary hospital for marrow hematopoietic progenitor cell donation. He did not have a history of transfusion. His blood type was A, RhD+, and findings from the antibody screening and identification test showed 2+ reactivity in all panel cells except autologous cells. We concluded that it would be very difficult to find compatible blood components for the donor and requested further tests from external laboratories. Anti-Ge2 was identified by additional tests in a foreign reference laboratory, and the Duffy genotype of the donor was FY*02/FY*02N.01 based on the Korean Rare Blood Program. Although the donor was not a Korean, as the number of foreign patients visiting Korea increases annually, there is growing interest in patients with rare blood types in the Korean population. However, there has been very little research on rare or high prevalence blood type antigen and antibody in the Korean population. Therefore, additional research in Korea is needed on rare blood group antibodies and antigens, including Ge cases.

Case of Acute Hemolytic Transfusion Reaction due to Anti-Fy(a) Alloantibody in a Patient with Autoimmune Hemolytic Anemia / 대한수혈학회지

A 72-year-old man with general weakness visited the outpatient clinic of the hematology department. The patient had been treated under the diagnosis of autoimmune hemolytic anemia for 2 years. His hemoglobin level at the time of the visit was 6.3 g/dL, and a blood transfusion was requested to treat his anemia. The patient's blood type was A, RhD positive. Antibody screening and identification test showed agglutination in all reagent cells with a positive reaction to autologous red blood cells (RBCs). He had a prior transfusion history with three least incompatible RBCs. The patient returned home after receiving one unit of leukoreduced filtered RBC, which was the least incompatible blood in the crossmatching test. After approximately five hours, however, fever, chills, dyspnea, abdominal pain, and hematuria appeared and the patient returned to the emergency room next day after the transfusion. The anti-Fy(a) antibody, which was masked by the autoantibody, was identified after autoadsorption using polyethylene glycol. He was diagnosed with an acute hemolytic transfusion reaction due to anti-Fy(a) that had not been detected before the transfusion. In this setting, it is necessary to consider the identification of coexisting alloantibodies in patients with autoantibodies and to become more familiar with the method of autoantibody adsorption.

A Case of Sickle Cell Anemia with a Lack of High Frequency Red Blood Cell Antigen / 대한수혈학회지

Patients with sickle cell anemia are chronically transfused. Therefore, it is important to prevent the alloimmunization of RBC antigens. The authors identified a high frequency antigen-negative blood group in patients with sickle cell anemia. As the number of foreigners residing in Korea is increasing, it is necessary to know what to consider when transfusing blood to sickle cell anemia patients. Patients with sickle cell anemia should be informed of the exact blood group type using extended RBC typing to confirm the ABO, Rh, Kell, and Duffy blood types at diagnosis or before the first blood transfusion. Extended matched blood transfusion can reduce the risk of alloimmunization of RBC antigens.

Survey and study on FYES and FYX alleles of Duffy blood group in Nanjing Han population / 国际检验医学杂志

Objective To investigate the FYES and FYX allele expression situation of Duffy blood group in Han population of Nanjing area.Methods The genomic DNA was randomly extracted from peripheral EDTA-K2 anticoagulation blood samples in 200 voluntary blood donors and Duffy blood group FYA and FYB allele typing was performed by PCR-SSP.The EYES and FYX allele sequence charateristics of Duffy blood group were analyzed by adopting Sanger double deoxygenation gene sequencing method.Results The PCR genotyping results were consistent with the sequencing results.Among 200 samples,180 cases were FYA/FYA and 20 cases were FYA/FYB;the sequencing results showed that FYES and FYX allele expression was not been found in 200 samples.Conclusion The expression frequencies of FYES and FYX alleles are extremely low in Han population of Nanjing area,which has significant difference compared with other races.

Duffy Blood Group Genotyping in Thai Blood Donors

BACKGROUND: Duffy (FY) blood group genotyping is important in transfusion medicine because Duffy alloantibodies are associated with delayed hemolytic transfusion reactions and hemolytic disease of the fetus and newborn. In this study, FY allele frequencies in Thai blood donors were determined by in-house PCR with sequence-specific primers (PCR-SSP), and the probability of obtaining compatible blood for alloimmunized patients was assessed. METHODS: Five hundred blood samples from Thai blood donors of the National Blood Centre, Thai Red Cross Society, were included. Only 200 samples were tested with anti-Fy(a) and anti-Fy(b) using the gel technique. All 500 samples and four samples from a Guinea family with the Fy(a-b-) phenotype were genotyped by using PCR-SSP. Additionally, the probability of obtaining antigen-negative red blood cells (RBCs) for alloimmunized patients was calculated according to the estimated FY allele frequencies. RESULTS: The FY phenotyping and genotyping results were in 100% concordance. The allele frequencies of FY*A and FY*B in 500 central Thais were 0.962 (962/1,000) and 0.038 (38/1,000), respectively. Although the Fy(a-b-) phenotype was not observed in this study, FY*B(ES)/FY*B(ES) was identified by PCR-SSP in the Guinea family and was confirmed by DNA sequencing. CONCLUSIONS: Our results confirm the high frequency of the FY*A allele in the Thai population, similar to that of Asian populations. At least 500 Thai blood donors are needed to obtain two units of antigen-negative RBCs for the Fy(a-b+) phenotype.

The Duffy binding protein as a key target for a Plasmodium vivax vaccine: lessons from the Brazilian Amazon

Plasmodium vivax infects human erythrocytes through a major pathway that requires interaction between an apical parasite protein, the Duffy binding protein (PvDBP) and its receptor on reticulocytes, the Duffy antigen/receptor for chemokines (DARC). The importance of the interaction between PvDBP (region II, DBPII) and DARC to P. vivax infection has motivated our malaria research group at Oswaldo Cruz Foundation (state of Minas Gerais, Brazil) to conduct a number of immunoepidemiological studies to characterise the naturally acquired immunity to PvDBP in populations living in the Amazon rainforest. In this review, we provide an update on the immunology and molecular epidemiology of PvDBP in the Brazilian Amazon - an area of markedly unstable malaria transmission - and compare it with data from other parts of Latin America, as well as Asia and Oceania.

Duffy blood group genotypes among malaria Plasmodium vivax patients of Baoulch population in Southeastern Iran

OBJECTIVE@#To determine the distribution of Duffy blood group genotypes in Balouch population as a major ethnic group that living in a sub-tropical area in south East of Iran.@*METHODS@#In this study, the Duffy blood group FY phenotypes were determined using indirect anti-globulin technique and also genotype by PCR-RFLP in 160 vivax malaria patients and 160 control individuals.@*RESULTS@#The results showed that the most common Duffy genotype was FYA/FYB (46.6%) followed by FYA/FYA (15.3%), FYA/FYO (14.4%), FYB/FYO (11.9%), FYB/FYB (10%) and FYO/FYO (1.9%). In case individuals, frequency of FYA, FYB and FYO alleles were 0.471, 0.431 and 0.097, respectively compaired to 0.444, 0.353 and 0.203, respectively in control (non-infected) group.@*CONCLUSIONS@#This data provide evidence that individuals with the FYA/FYB genotype have higher susceptibility to malaria and there are significant associations between Duffy blood group variants and susceptibility or resistance to vivax malaria.

Expresión del fenotipo Duffy negativo en mujeres afrodescendientes y su relación con la preeclampsia / Expression of the Duffy negative phenotype in women of African descent and their relation to pre-eclampsia

La preeclampsia (PE) es una complicación del embarazo que trae consigo algunas consecuencias negativas para la madre y el feto: en la madre provoca principalmente hipertensión y proteinuria, mientras que en el feto puede presentarse trombocitopenia, alteración en el desarrollo del sistema nervioso central y circulatorio, y restricción del crecimiento intrauterino, lo cual se considera el factor de riesgo principal de muerte fetal en nacimientos producto de una PE severa. En la preeclampsia se presenta una disfunción endotelial relacionada con placentación anormal, estado de estrés oxidativo y proceso inflamatorio sistémico, que lleva a la activación de neutrófilos y monocitos. Se ha considerado a la interleucina-8 (IL-8) como un posible candidato desencadenante por ser quimioatrayente y activador de leucocitos; en la circulación sanguínea, la IL-8 se une a un receptor de quimiocina multiespecífico de alta afinidad denominado DARC, que es idéntico al antígeno del grupo sanguíneo Duffy. Este receptor regula los niveles plasmáticos de IL-8, uniéndose a esta quimiocina, pero cuando hay una mutación en la región promotora del gen se altera la expresión de DARC, lo que conlleva a que la IL-8 de los factores genéticos involucrados en la activación de los neutrófilos y de los monocitos, y por ende, en la disfunción endotelial presentada durante este síndrome hipertensivo, especialmente en la población afrodescendiente.

Preeclampsia (PE) is a complication of pregnancy that brings some negative consequences for both mother and fetus. It specially causes hypertension and proteinuria in mothers; while in fetuses it causes thrombocytopenia, development alterations of the central nervous and circulatory system; also intrauterine growth restriction may occur. This last factor is regarded as the main risk factor for fetal death in births as a result of severe PE. There is endothelial dysfunction in preeclampsia related to abnormal placentation, state of oxidative stress and systemic inflammatory process that leads to the activation of neutrophils and monocytes. Interleukin-8 (IL-8) is considered as a possible trigger candidate, since this chemokine is a chemoattractant and leukocyte activator. In the bloodstream, interleukin-8 binds to a high affinity multispecific-chemokine receptor called DARC, which is identical to the Duffy blood group antigen. This receptor regulates plasma levels of IL-8 by binding to chemokine. But, when there is a mutation in the gene promoter region, DARC expression is altered, and IL-8 inefficiently binds to receptor. This mutation results in Duffy negative phenotype, which is present in most of African descendants. This literature review is intended to address the role of IL-8 as neutrophil chemo-attractant, the importance of Duffy blood system and the possible association between ethnicity and preeclampsia.

Characteristics of Duffy Blood Group Antigens and Their Global Distribution / 대한수혈학회지

The Duffy antigen was discovered in 1950, in a multiply transfused hemophiliac. Important progress has since been made in understanding the Duffy blood group system and its complexity. The Duffy blood group antigen (gp-Fy) is present primarily in erythrocytes, and also in endothelial cells of capillary and postcapillary venules, Purkinje cells of cerebellum, kidney, and pulmonary alveoli. The gp-Fy serves not only as a blood group antigen, but also as a receptor for chemokines, and as a receptor for Plasmodium vivax malaria parasites. The Duffy antigen is encoded by the DARC gene, its approved name is Duffy blood group chemokine receptor. Investigation of the DARC gene can help us in understanding the relationship of infectious disease to race or population. In addition, the allelic frequency of DARC varies according to the geographic area, which appears to reflect the history that mankind had adapted to environments and diseases, emigrating. As a result, further study of Duffy antigens can provide us with an integral and sound understanding of the human race.

High-Resolution Melting Analysis for Genotyping Duffy Blood Group Antigens / 대한수혈학회지

BACKGROUND: Accurate typing of Duffy blood group is important because anti-Duffy antibodies cause hemolytic transfusion reaction and hemolytic disease of the newborn. The aim of this study was to evaluate a new genotyping method using high resolution melting (HRM) analysis, a rapid and inexpensive approach for high-throughput Duffy genotyping. METHODS: A total of 20 unrelated Korean blood samples were obtained and an African-black sample was used for GATA control. Phenotyping was performed by hemagglutination (DiaMed AG, Switzerland). GATA and FYA/B PCR products were obtained by PCR-restriction fragment length polymorphism (RFLP) using Taq DNA polymerase (Promega, WI) and enzymes BanI and StyI (New England Biolab, UK). For HRM, PCR amplification was performed using LightCycler 480 ResoLight Dye (Roche, USA) and Lightcycer 480 (Roche, USA). RESULTS: Phenotyping and genotyping data using PCR-RFLP and HRM analysis were compared. Different types of HRM curves were obtained according to genotypes, FYA/FYA, FYB/FYB, and FYA/FYB, and to GATA mutations, homozygote FYB-33T (T/T), heterozygote FYB-33T/33C (T/C), and homozygote FYB-33C (C/C). Phenotypes 18 Fy(a+b-), 1 Fy(a+b+), 1 Fy(a-b+), and 1 Fy(a-b-) showed complete concordance with genotyping methods. Fy(a-b-) sample was found to be a FYB-33C homozygote by both genotyping methods. CONCLUSION: Phenotyping and genotyping showed concordant results and both genotyping methods using PCR-RFLP and HRM analysis showed good agreement in finding mutation in GATA and FY gene coding regions. HRM analysis is suitable and reliable for high-throughput screening for Duffy genotyping.

Blood group phenotype frequencies in blood donors from a tertiary care hospital in north India

BACKGROUND: Knowledge about the frequency of red blood cell-antigen phenotypes in a population can be helpful in the creation of a donor data bank for the preparation of indigenous cell panels and for providing antigen-negative compatible blood to patients with multiple alloantibodies. METHODS: ABO and RhD blood grouping was performed on 9,280 continuous voluntary and replacement donors. For other rare blood groups, 508 ACD blood samples were obtained from the donors at the Blood Bank of the Department of Transfusion Medicine, All India Institute of Medical Sciences (AIIMS), New Delhi, India. Blood group antigens were determined by tube method using anti-sera (Bio-Rad, USA), and the phenotype frequencies were expressed as percentages. RESULTS: Group B (37.39%) was the most common, followed by group O (31.85%). R1R1 and rr were the most common phenotypes amongst Rh positive and Rh negative groups, respectively. A rare phenotype R2Rz was found in one donor. For Kidd and Duffy blood group systems, Jk (a+b+) and Fy (a+b+) were the most common phenotypes (46.06% and 48.03%, respectively). The most common phenotypes for MNSs, Lu, and Kell blood groups were M+N+, S-s+, Lu (a-b+), and K-k+, respectively. A very rare case of Fy (a-b-) and Jk (a-b-) was found in a single donor. CONCLUSION: This study is the first small step to create a rare donor data bank and to prepare indigenous cell panels to provide compatible blood to all multi-transfused alloimmunized patients.

Blood group phenotype frequencies in blood donors from a tertiary care hospital in north India

BACKGROUND: Knowledge about the frequency of red blood cell-antigen phenotypes in a population can be helpful in the creation of a donor data bank for the preparation of indigenous cell panels and for providing antigen-negative compatible blood to patients with multiple alloantibodies. METHODS: ABO and RhD blood grouping was performed on 9,280 continuous voluntary and replacement donors. For other rare blood groups, 508 ACD blood samples were obtained from the donors at the Blood Bank of the Department of Transfusion Medicine, All India Institute of Medical Sciences (AIIMS), New Delhi, India. Blood group antigens were determined by tube method using anti-sera (Bio-Rad, USA), and the phenotype frequencies were expressed as percentages. RESULTS: Group B (37.39%) was the most common, followed by group O (31.85%). R1R1 and rr were the most common phenotypes amongst Rh positive and Rh negative groups, respectively. A rare phenotype R2Rz was found in one donor. For Kidd and Duffy blood group systems, Jk (a+b+) and Fy (a+b+) were the most common phenotypes (46.06% and 48.03%, respectively). The most common phenotypes for MNSs, Lu, and Kell blood groups were M+N+, S-s+, Lu (a-b+), and K-k+, respectively. A very rare case of Fy (a-b-) and Jk (a-b-) was found in a single donor. CONCLUSION: This study is the first small step to create a rare donor data bank and to prepare indigenous cell panels to provide compatible blood to all multi-transfused alloimmunized patients.