Association of blood groups/Rh and diabetes mellitus in Karachi city, Pakistan / Associação de grupos sanguíneos/Rh e diabetes mellitus na cidade de Karchi, Paquistão

Abstract The main purpose of this study was to find out a possible association between ABO blood groups or Rh and diabetes mellitus (DM) in the local population of eight (8) different towns of Karachi, Pakistan. For this purpose a survey was carried out in Karachi to have a practical observation of these towns during the period of 9 months from June 2019 to Feb. 2020. Out of eighteen (18) towns of Karachi, samples (N= 584) were collected from only eight (8) Towns of Karachi and gave a code-number to each town. Diabetic group sample was (n1=432) & pre-diabetes sample was (n2 =152). A standard Abbot Company Glucometer for Random Blood Sugar (RBS) and Fasting Blood Sugar (FBS) tests, standard blood anti sera were used for ABO/Rh blood type. Health assessment techniques were performed ethically by taking informed consent from all registered subjects. Finally data was analyzed by SPSS version 20.0. In our current study, the comparison of ABO blood groups frequencies between diabetic and pre-diabetic individuals were carried out. The percentage values of blood Group-B as given as: (32% in DM vs. 31% in pre-diabetics), followed by blood Group-O as: (18% in DM vs. 11% in pre-diabetics). Contrary to Group-"B" & "O", blood Group-A and Group-AB were distribution percentage higher pre-diabetic as compared to DM patients, as given as: Group-A (32% in pre-diabetics vs. 26% in DM) & Group-AB (26% in pre-diabetics vs. 24% in diabetic's patients). In addition, percentage distribution of Rh system was also calculated, in which Rh+ve Group was high and more common in DM patients as compared to pre-diabetics; numerically given as: Rh+ve Group (80% in DM vs. 72% in pre-diabetics). Different views and dimensions of the research topic were studied through literature support, some have found no any association and some established a positive association still some were not clear in making a solid conclusion. It is concluded that DM has a positive correlation with ABO blood groups, and people with Group-B have increased susceptibility to DM disease.

Resumo O objetivo principal deste estudo foi descobrir uma possível associação entre grupos sanguíneos ABO ou Rh e diabetes mellitus (DM) na população local de oito (8) diferentes cidades de Karachi, Paquistão. Para tanto, foi realizado um levantamento em Karachi para observação prática dessas cidades durante o período de 9 meses de junho de 2019 a fevereiro de 2020.De dezoito (18) cidades de Karachi, as amostras (N = 584) foram coletadas de apenas oito (8) cidades de Karachi e deram um número-código para cada cidade. A amostra do grupo de diabéticos foi (n1 = 432) e a amostra de pré-diabetes foi (n2 = 152). Um glicômetro padrão da Abbot Company para testes de açúcar no sangue aleatório (RBS) e açúcar no sangue em jejum (FBS), antissoros de sangue padrão foram usados ​​para o tipo de sangue ABO / Rh. As técnicas de avaliação de saúde foram realizadas de forma ética, tomando o consentimento informado de todos os indivíduos registrados. Finalmente, os dados foram analisados ​​pelo SPSS versão 20.0.No presente estudo, foi realizada a comparação das frequências dos grupos sanguíneos ABO entre diabéticos e pré-diabéticos. Os valores percentuais do sangue do Grupo-B são dados como: (32% em DM vs. 31% em pré-diabéticos), seguido pelo sangue do Grupo-O como: (18% em DM vs. 11% em pré-diabéticos). Ao contrário dos Grupos "B" e "O", sangue do Grupo-A e Grupo-AB tiveram distribuição percentual maior de pré-diabéticos em comparação com pacientes com DM, dado como: Grupo-A (32% em pré-diabéticos vs. 26% em DM) e Grupo AB (26% em pré-diabéticos vs. 24% em pacientes diabéticos). Além disso, também foi calculada a distribuição percentual do sistema Rh, no qual o Grupo Rh + ve foi elevado e mais comum em pacientes com DM em comparação aos pré-diabéticos; dados numericamente como: Grupo Rh + ve (80% em DM vs. 72% em pré-diabéticos). Diferentes visões e dimensões do tema de pesquisa foram estudadas com o suporte da literatura, alguns não encontraram nenhuma associação e alguns estabeleceram uma associação positiva, embora alguns não estivessem claros em fazer uma conclusão sólida. Conclui-se que o DM tem correlação positiva com os grupos sanguíneos ABO, e as pessoas com o Grupo B têm maior suscetibilidade à doença DM.

Genotipificación del gen RHD en ADN fetal libre en plasma de embarazadas RhD negativo / Genotyping of the RHD gene in free fetal DNA in plasma of RhD negative pregnant women

Objetivo: Determinar el grupo RhD fetal a través del estudio del gen RHD en ADN fetal que se encuentra libre en plasma de embarazadas RhD negativo. Método: Se analizó la presencia de los genes RHD, SRY y BGLO en ADNfl obtenido de plasma de 51 embarazadas RhD negativo no sensibilizadas, utilizando una qPCR. Los resultados del estudio genético del gen RHD se compararon con el estudio del grupo sanguíneo RhD realizado por método serológico en muestras de sangre de cordón, y los resultados del estudio del gen SRY fueron cotejados con el sexo fetal determinado por ecografía. Se calcularon la sensibilidad, la especificidad, los valores predictivos y la capacidad discriminativa del método estandarizado. Resultados: El gen RHD estaba presente en el 72,5% de las muestras y el gen SRY en el 55,5%, coincidiendo en un 100% con los resultados del grupo RhD detectado en sangre de cordón y con el sexo fetal confirmado por ecografía, respectivamente. Conclusiones: Fue posible deducir el grupo sanguíneo RhD del feto mediante el estudio del ADN fetal que se encuentra libre en el plasma de embarazadas con un método molecular no invasivo desarrollado y validado para este fin. Este test no invasivo puede ser utilizado para tomar la decisión de administrar inmunoglobulina anti-D solo a embarazadas RhD negativo que portan un feto RhD positivo.

Objective: To determine the fetal RhD group through the study of the RHD gene in fetal DNA found free in plasma of RhD negative pregnant women. Method: The presence of the RHD, SRY and BGLO genes in fetal DNA obtained from plasma of 51 non-sensitized RhD negative pregnant women was analyzed using qPCR. The results of the genetic study of the RHD gene were compared with the RhD blood group study performed by serological method in cord blood samples, and the results of the SRY gene study were compared with the fetal sex determined by ultrasound. Sensitivity, specificity, predictive values and discriminative capacity of the standardized method were calculated. Results: The RHD gene was present in 72.5% of the samples and the SRY gene in 55.5%, coinciding 100% with the results of the RhD group detected in cord blood, and with the fetal sex confirmed by ultrasound, respectively. Conclusions: It was possible to deduce the RhD blood group of the fetus through the study of fetal DNA found free in the plasma of pregnant women with a non-invasive molecular method developed and validated for this purpose. This non-invasive test can be used to make the decision to administer anti-D immunoglobulin only to RhD-negative pregnant women carrying an RhD-positive fetus.

Analysis of Irregular Blood Group Antibody Distribution and Blood Transfusion Efficacy in Patients with Malignant Tumor / 中国实验血液学杂志

OBJECTIVE@#To investigate the distribution of irregular blood group antibodies in patients with malignant tumors, and to analyze the relationship between it and efficacy of blood transfusion in patients.@*METHODS@#5 600 patients with malignant tumors treated in Shanxi Bethune Hospital from January 2019 to December 2021 were selected as the research subjects. All patients received blood transfusion, and cross matching test was conducted before blood transfusion, irregular antibody results of patients were tested; the irregular distribution of blood group antibodies was observed, and the relationship between it and efficacy of blood transfusion in patients was analyzed.@*RESULTS@#Among 5 600 patients with malignant tumors, 96 cases were positive for irregular antibody, and the positive rate was 1.71%; the main blood group systems involved in the irregular antibody positive of 96 patients with malignant tumors were RH, MNSs and Duffy system, among which Rh blood group was the most common, and the proportion of anti-E was the highest; among the malignant tumor patients with positive blood group irregular antibody, the proportion of female was higher than that of male; the proportion of patients aged >60 years was the highest, followed by patients aged >40 and ≤50 years, and the proportion of patients aged 18-30 years was the lowest; the patients with positive blood group irregular antibody were mainly in blood system (including lymphoma), digestive system, reproductive and urinary system; the positive rate of irregular antibody of patients in the ineffective group was higher than that of patients in the effective group, the difference was statistically significant (P<0.05). Logistic regression analysis results showed that, irregular antibody positive was a risk factor for ineffective blood transfusion in patients with malignant tumor (OR>1, P<0.05).@*CONCLUSION@#The irregular blood group antibody positive of patients with malignant tumor are mostly female, and the proportion of patients aged >60 is the highest, which is mainly distributed in malignant tumors of blood system, digestive system and urogenital system, and the positive blood group irregular antibody is related to the efficacy of blood transfusion in patients.

RHD Gene Analysis of A Blood Donor with Del Phenotype / 中国实验血液学杂志

OBJECTIVE@#To analyze the RHD genotype of a blood donor with Del phenotype in Yunnan.@*METHODS@#Rh serological phenotype was identified. RHD gene was detected by PCR-SSP typing, and its 10 exons were sequenced. Exon 9 was amplified for sequencing and analysis. RHD zygosity was detected.@*RESULTS@#The Rh phenotype of this specimen was CcDelee. Genomic DNA exhibited a 1 003 bp deletion spanning from intron 8, across exon 9 into intron 9. The deletion breakpoints occurred between two 7-bp short tandem repeat sequences. There was no variation in the sequences of the remaining exons. The Rh hybridization box test showed that there was one RHD negative allele.@*CONCLUSION@#This specimen is Del type caused by deletion of RHD exon 9.

Análise do perfil de Rh e Kell em doadores de sangue em Porto Alegre, Rio Grande do Sul / Phenotypic profile analysis of Rh and Kell blood group systems among donors in Porto Alegre, Rio Grande do Sul

Introdução: A hemoterapia é uma prática terapêutica pelo meio de transfusão sanguínea. Devido ao baixo estoque de bolsas de sangue e o aumento de pacientes crônicos e emergenciais, se faz necessária a realização de testes imuno-hematológicos para minimizar os riscos de reações transfusionais e aloimunizações em doadores e receptores de sangue. Deste modo, no estudo foi avaliada a prevalência dos antígenos dos sistemas Rh e Kell em doadores de sangue de Porto Alegre ­ RS.Métodos: Estudo quantitativo, transversal e retrospectivo que foi realizado através da análise das informações dos doadores de sangue contidas no banco de dados do Hemocentro do Estado do Rio Grande do Sul, nos anos de 2018 e 2019.Resultados: Das 6.479 amostras fenotipadas, quanto ao sistema Rh, 44,6% são Rh positivo e 55,4% são Rh negativo. As frequências dos antígenos encontradas foram de, CC 10,1%, Cc 27%, cc 62,9%, EE 1,2%, Ee 13,9%, ee 84,9%. E, para o sistema Kell, K1 positivo 7,1% e K1 negativo 92,9%.Conclusões: Antígenos do sistema Rh e Kell exibem um grande nível de imunogenicidade e uma forte ligação com a Doença Hemolítica do Recém-nascido, podendo ocorrer a sensibilização em pacientes caso não haja a compatibilidade sanguínea. Este estudo ressalta a importância da implementação da fenotipagem eritrocitária em doadores de sangue, sugere-se mais estudos com períodos distintos para a pesquisa de resultados satisfatórios.

Introduction: Hemotherapy is a therapeutic practice consisting of blood transfusion. Low blood supply and an increase in chronic and emergency patients have made it necessary to conduct immunohematology tests to minimize the risks of adverse reactions and alloimmunization in donors and recipients. Therefore, this study aimed to assess the prevalence of Rh and Kell blood group antigens among blood donors in Porto Alegre, Rio Grande do Sul, Brazil.Methods: We conducted a quantitative, cross-sectional, retrospective study. Information from blood donors included in the Rio Grande do Sul's Blood Center database from 2018 to 2019 were analyzed.Results: A total of 6,479 samples were phenotyped, of which 44.6% were Rh-positive and 55.4% were Rh-negative. Antigen prevalence was CC (10.1%), Cc (27%), cc (62.9%), EE (1.2%), Ee (13.9%), and ee (84.9%). As for the Kell group, 7.1% were K1-positive and 92.9% were K1-negative.Conclusions: The Rh and Kell antigens are highly immunogenic and have a strong link with the hemolytic disease of the newborn. Sensitization may occur in patients if there is no blood compatibility. This study highlights the importance of implementing erythrocyte phenotyping in blood donors. Further studies should be conducted in different time frames to achieve satisfactory results.

Identification of Weak D Type 1 in Rh Blood Group System and Discussion of Transfusion Strategy / 中国实验血液学杂志

OBJECTIVE@#To investigate the molecular mechanism of one patient with abnormal serological phenotype in RhD and discuss the transfusion strategy.@*METHODS@#The RhD variant sample was screened from a patient with IgM type anti-D antibody and further determined by three different sources of anti-D antibodies. Ten exons and the adjacent introns of the RHD gene were amplified, purified and sequenced. RhCE phenotypes and RHCE genotypes were detected.@*RESULTS@#The patient with Rh variant showed abnormal results of serological tests. The RHD gene sequence analysis showed that the RHD*01W.01 with a variation (c.809T>G, p.Val270Gly) in exon 6 of the RHD gene was found in the patient. The RhCE phenotype was CcEe. The genotyping results of RHCE were consistent with the serological typing results.@*CONCLUSION@#The Rh variant of the patient is RHD*01W.01, these findings indicate that RhD variants should be analyzed by molecular assays for the sake of safe transfusion.

Prevalencia de anquiloglosia en neonatos y relación con datos auxológicos del recién nacido o con otras malformaciones o enfermedades asociadas / Prevalência de anquiloglossia em recém-nascidos e relação com dados auxológicos do recém-nascido ou com outras malformações ou doenças associadas / Prevalence of ankyloglossia in neonates and relationship with auxological data of the newborn or with other malformations or associated diseases

Objetivo: determinar la prevalencia de anquiloglosia en neonatos del Hospital Nostra Senyora de Meritxell del Principado de Andorra, Europa, así como la relación existente con datos auxológicos u otras malformaciones o enfermedades asociadas. Material y métodos: Se realizó un estudio descriptivo, transversal, retrospectivo poblacional de los recién nacidos durante un período de 8 meses. Se estudiaron once variables: presencia o no de anquiloglosia, tipo de frenillo corto, talla, peso, perímetro craneal, sexo, Rh y grupo sanguíneo, edad gestacional, malformaciones y patologías neonatales asociadas, de forma confidencial y anónima, correlacionándolas para ver si existía alguna asociación entre ellas. Resultados: Un total de 306 recién nacidos fueron incluidos en el estudio (52% varones y 48% mujeres). La prevalencia de anquiloglosia fue del 6,54% (n=20). Del total de varones (159/306), el 8,125% (n=13) presentaban anquiloglosia, mientras que en mujeres (147/306), la prevalencia fue del 4,79% (n=7), sin encontrar diferencias estadísticamente significativas respecto al sexo (p-valor=0,24). Según la clasificación de Coryllos, el tipo II fue el más frecuente (95% de los casos) y el 4,58% de los lactantes con anquiloglosia exhibieron patología/malformación. Conclusiones: La prevalencia de anquiloglosia en recién nacidos en Andorra, Europa, es similar a la observada por otros autores en investigaciones similares. No se observó asociación entre la presencia o no anquiloglosia y las variables estudiadas.

Objetivo: determinar a prevalência de anquiloglossia em recém-nascidos do Hospital Nostra Senyora de Meritxell do Principado de Andorra, Europa, bem como a relação existente com dados auxológicos ou outras malformações ou doenças associadas. Material e métodos: Foi realizado um estudo descritivo, transversal e retrospectivo da população de recém-nascidos durante um período de 8 meses. Onze variáveis foram estudadas: presença ou não de anquiloglossia, tipo de frênulo curto, altura, peso, perímetro craniano, sexo, Rh e grupo sanguíneo, idade gestacional, malformações e patologias neonatais associadas, de forma confidencial e anônima, correlacionando-as para ver se existia alguma associação entre eles. Resultados: Um total de 306 recém-nascidos foram incluídos no estudo (52% masculino e 48% feminino). A prevalência de anquiloglossia foi de 6,54% (n=20). Do total de homens (159/306), 8.125% (n=13) apresentavam anquiloglossia, enquanto nas mulheres (147/306) a prevalência foi de 4,79% (n=7), sem encontrar diferenças estatisticamente significantes em relação ao sexo (valor-p = 0,24). De acordo com a classificação de Coryllos, o tipo II foi o mais frequente (95% dos casos) e 4,58% dos lactente com anquiloglossia apresentaram patologia / malformação. Conclusões: A prevalência de anquiloglossia em recém-nascidos em Andorra, Europa, é semelhante à observada por outros autores em investigações semelhantes. Não foi observada associação entre a presença ou não anquiloglossia e as variáveis estudadas

Aim: to determine the prevalence of ankyloglossia in new-borns of the Nostra Senyora de Meritxell Hospital in the Principado de Andorra, Europe, as well as the existing relationship with auxological data or other malformations or associated diseases. Material and methods: A descriptive, cross-sectional, retrospective population study of 306 newborns was carried out over a period of 8 months. Eleven variables were studied: presence or no of ankyloglossia, type of short frenulum, height, weight, cranial perimeter, sex, Rh and blood group, gestational age, malformations and associated neonatal pathologies, confidentially and anonymously, correlating them to see if there was any association between them. Results: A total of 306 new-borns were included in the study (52% male and 48% female). The prevalence of ankyloglossia was 6,54% (n=20). Of the total of men (159/306), 8,125% (n=13) presented ankyloglossia, while woman (147/306), the prevalence was 4,79% (n=7), without finding statistically significant differences regarding sex (p-value=0,24). According to the Coryllos classification, type II was the most frequent (95% of cases) and 4,58% of infants with ankyloglossia exhibited pathology/malformation. Conclusions: The prevalence of ankyloglossia in new-borns in Andorra, Europe, is similar to that observed by other authors in similar research. No association was observed between the presence or not of ankyloglossia and the variables studied

Detection of Irregular Antibodies after Blood Transfusion for Children with Thalassemia in Hainan / 中国实验血液学杂志

OBJECTIVE@#To investigate the irregular antibody positive rate and antibody specificity in children with thalassemia received long-term blood transfusion in Hainan area and analyze the causes of antibody screening positive.@*METHODS@#Micro-column gel method was used to screen the irregular antibody in 49 children who received transfusion treatment in our hospital, and the antibody specificity of the positive samples was evaluated.@*RESULTS@#Fourteen of 49 cases showed positive for screening. Among them, 11 cases showed Rh blood group antibody after detecting antibody specificity, 1 case showed the coexistence of irregular antibody and autoantibody. One case for anti-JK@*CONCLUSION@#Most of the antibodies produced after long-term blood transfusion in the children with thalassemia belong to Rh blood group antibodies; the children with mixed thalassemia are more likely to produce antibodies; the antibody screening positive rate of Li nationality is higher than that of Han nationality, which may be caused by the genetic difference of blood type between Li nationality and Han nationality.

Molecular genetic analysis of two individuals with weak D variant of the Rh blood type / 中华医学遗传学杂志

OBJECTIVE@#To explore the molecular basis of two individuals with weak D variant of the Rh blood type.@*METHODS@#Routine serological testing was carried out to detect the D, C, c, E and e antigens of the Rh blood group. The D antigen was further detected with an indirect antiglobulin test. The presence of Rhesus box was detected by PCR to determine the homozygosity of the RHD gene.@*RESULTS@#Both samples were determined as weak D phenotype by the indirect antiglobulin test. DNA sequencing revealed that case 1 harbored a heterozygous 208C>T variant in exon 2 and a heterozygous 1227G>A variant in exon 9; while case 2 harbored homozygous 779A>G variants of exon 5 of the RHD gene. Case 1 was determined as RHD+/RHD+, while case 2 was determined as RHD+/RHD-. The two samples were respectively named as weak D type 122 and weak D type 149 based on the rules of Rhesus Base Nomenclature.@*CONCLUSION@#D negative blood donors should subject to indirect antiglobulin testing and molecular analysis for safer transfusion.

A pedigree analysis of a rare RhD 336-1G>A intron variant / 中华医学遗传学杂志

OBJECTIVE@#To explore the molecular mechanism of a case where RhD genotyping did not match serological results.@*METHODS@#The serological results of 8 members from two generations of this family were analyzed. And according to Mendelian law of inheritance, RhD genotyping, zygotic type determination and gene sequencing were performed for the family members.@*RESULTS@#The proband and one of her cousins have the same RhD alleles, both of them have a 336-1G>A intron variant RhD allele and a complete RhD deletion allele. The variant alleles are inherited from two of their parents with blood relationship, while the complete-deleted alleles come from the other. 336-1G>A means that the last base G of the second intron of the RhD gene is mutated to A, which leads to a negative RhD serology and a positive genotype in the proband.@*CONCLUSION@#There was a rare 336-1G> A intron variant gene (RhD * 01N.25) in this family, which was a recessive gene relative to the RhD gene and resulted in RhD phenotype negative.

The Blood Group Serology Combined Irregular Antibody Prenatal Screening Strategies for RhD / 中国实验血液学杂志

OBJECTIVE@#To investigate the distribution of Rh phenotype, irregular antibodies screenting and postpartum follow-up of RhD@*METHODS@#The samples of 313 RhD@*RESULTS@#The Rh serological phenotypes of 313 RhD@*CONCLUSION@#The Rh serological phenotype of RhD

Serologic strategy in detecting RHD altered alleles in Brazilian blood donors

ABSTRACT Background: We evaluated different technological approaches and anti-D clones to propose the most appropriate serologic strategy in detecting the largest numbers of D variants in blood donors. Methods: We selected 101 samples from Brazilian blood donors with different expressions of D in our donor routine. The tests were performed in immediate spin (IS) with eleven commercially available anti-D reagents in a tube and microplate. The D confirmatory tests for the presence of weak D included the indirect antiglobulin test (IAT) in a tube, gel and solid-phase red blood cell adherence (SPRCA). All DNA samples were extracted from peripheral blood and the D variants were classified using different molecular assays. Results: The RHD variants identified by molecular analysis included weak D types (1, 2, 3, 11 and 38) and partial Ds (DAR1.2, DAR1, DAR3.1, DAU0, DAU2, DAU4, DAU5, DAU6, DMH and DVII). The monoclonal-monoclonal blend RUM-1/MS26 was the best anti-D reagent used in detecting the D antigen in the IS phase in a tube, reacting with 83.2% of the D variants, while the anti-D blend D175 + 415 was the best monoclonal antibody (MoAb) used in a microplate to minimize the need for an IAT, reacting with 83.2% of the D variants. The D confirmatory tests using SPRCA showed a reactivity (3 - 4+) with 100% of the D variant samples tested. Conclusion: Our results show that, even using sensitive methods and MoAbs to ensure the accurate assignment of the D antigen, at least 17% of our donor samples need a confirmatory D test in order to avoid alloimmunization in D-negative patients.

Sistema Rh-Hr y variantes del antígeno D en tres poblaciones afroecuatorianas del Valle del Chota / Rh-Hr system and D antigen variants in three Afro-Ecuadorian populations in the Chota Valley / Sistema Rh-Hr e variantes do antígeno D em três populações afro-equatorianas no vale do Chota

Resumen La identificación inequívoca del antígeno D en medicina transfusional es de vital importancia para evitar reacciones postransfusionales y la enfermedad hemolítica del recién nacido. Es común el uso de reactivos serológicos monoclonales o tarjetas de gel y su interpretación está definida por cruces, de acuerdo con la reacción serológica. El propósito de este estudio fue determinar la frecuencia del factor Rh y las variantes del antígeno D en una población afroecuatoriana. Se trató de un estudio descriptivo, transversal con muestreo aleatorio simple de 541 pobladores. Para la tipificación del factor Rh se utilizó la metodología en tubo con antisueros monoclonales y para la detección de las variantes de D se utilizaron tarjetas de gel IDCoombs Anti-IgG. Las lecturas se verificaron mediante el análisis del índice kappa. Se aplicó estadística descriptiva y el análisis de Chi cuadrado para establecer la relación de las variables y su significación. Se identificó una frecuencia del 92% de individuos Rh(D) positivo y un 8% Rh(D) negativo. El 4,80% de los individuos presentaban la variante D débil y el 79% reacciones serológicas entre 2 y 3(+) indicativas de otras variantes del antígeno D. El fenotipo más común fue el R0/R0. Estos datos demuestran la necesidad de confirmar la existencia de variantes del antígeno D en esta población para un mejor manejo de la sangre. Una limitante constituye la disponibilidad de técnicas moleculares para la genotipificación de D; sin embargo, se podría implementar la fenotipificación RHCE como estrategia pretransfusional.

Abstract The unequivocal identification of D antigen in transfusion medicine is of vital importance to avoid post-transfusion reactions and hemolytic disease of the newborn. The use of monoclonal serological reagents or gel cards is common and their interpretation is defined according to the serological reaction by crosses. The purpose of this study was to determine the frequency of Rh factor and D antigen variants in the Afro-Ecuadorian population. This was a descriptive, cross-sectional study with simple random sampling of 541 residents. Tube typing with monoclonal antisera was used to typify Rh factor and ID-Coombs Anti-IgG gel cards were used to detect D variants, and the readings were verified by analysis of the kappa index. Descriptive statistics and Chi-square analysis were applied for the relationship of the variables and their significance. A frequency of 92% of Rh(D) positive individuals and 8% Rh(D) negative individuals were identified. Almost 5% (4.80%) of the individuals presented the weak D variant and 79% serological reactions between 2-3(+) indicative of other D antigen variants, the most common phenotype being R0/R0. These data demonstrate the need to confirm the existence of D antigen variants in this population for better management and availability of blood. A limitation is the availability of molecular techniques for D genotyping, however, RHCE phenotyping could be implemented as a pretransfusion strategy.

Resumo A identificação inequívoca do antígeno D na medicina transfusional é de vital importância para evitar reações pós-transfusionais e a doença hemolítica do recém-nascido. É comum o uso de reagentes sorológicos monoclonais ou cartões de gel e sua interpretação é definida por cruzamentos de acordo com a reação sorológica. O objetivo deste estudo foi determinar a frequência do fator Rh e as variantes do antígeno D numa população afro-equatoriana. Foi um estudo descritivo, transversal, com amostragem aleatória simples de 541 residentes. Para a tipagem do fator Rh foi utilizada a metodologia em tubo com anti-soros monoclonais e para a detecção das variantes de D, os cartões de gel ID-Coombs Anti-IgG. As leituras foram verificadas por análise do índice kappa. Foi aplicada estatística descritiva e para estabelecer a relação das variáveis e sua significação se utilizou a análise do qui-quadrado. Identificando uma frequência de 92% dos indivíduos Rh (D) positivos e 8% Rh (D) negativos. 4,80% dos indivíduos apresentavam a variante D fraca e 79% reações sorológicas entre 2 e 3(+) indicativas de outras variantes do antígeno D, sendo o fenótipo mais comum o R0/R0. Esses dados demonstram a necessidade de confirmar a existência de variantes do antígeno D nessa população para melhor gerenciamento e disponibilidade de sangue. Uma limitação é a disponibilidade de técnicas moleculares para a genotipagem de D, no entanto, a fenotipagem de RHCE poderia ser implementada como uma estratégia de pré-transfusão.

Maternal intravenous immunoglobulin: a non-invasive treatment option for Rhesus D-sensitised women with previous adverse pregnancy outcomes

Background. Maternal intravenous immunoglobulin (IVIG) may delay the onset and severity of fetal anaemia in Rhesus D (RhD)- sensitised pregnancies, thereby minimising the need for intrauterine transfusion and its associated complications. Objective. To compare the pregnancy outcomes of RhD-sensitised women who received antenatal IVIG with those who did not receive antenatal IVIG. Methods. This was a retrospective cross-sectional analysis of RhD-sensitised women who attended the Wits Fetal Medicine Centre (Johannesburg) from 1 January 2008 to 31 May 2018. Criteria for maternal IVIG administration were: (i) previous adverse pregnancy outcome (early neonatal death, intrauterine fetal death or miscarriage related to RhD sensitisation), (ii) women with high antibody titre levels (≥1:64) in the absence of fetal anaemia; and (iii) rising antibody titre levels. Maternal antibody titre levels, pregnancy and neonatal outcomes were compared in women who received IVIG v. those who did not receive IVIG. Results. Of the 42 RhD-sensitised women, 14 received IVIG. A greater proportion of women experienced a decrease in antibody titres in the IVIG v. no-IVIG group (43% v. 11%, respectively; p=0.04). Nine of the 10 women in the IVIG group with a previous adverse pregnancy outcome had a successful pregnancy outcome following IVIG treatment. Conclusion. Maternal IVIG may provide a successful pregnancy outcome in RhD-sensitised women with previous adverse pregnancy outcomes related to Rh disease, or women with raised or increasing maternal antibody titre levels who present in the first or early second trimester

Procedencia y fenotipo del sistema Rh en donantes negativos en un Hemocentro colombiano / Origin and phenotype of the Rh system in negative donors in a Colombian hemocenter

percentIntroducción: El sistema Rh está compuesto por más de 56 antígenos capaces de producir enfermedad hemolítica, definidos por métodos serológicos. Los más importantes y, por ello denominados antígenos mayores del sistema, son los antígenos D, C, c, E y e. Estos antígenos se ubican sobre dos proteínas que se expresan en la membrana de los eritrocitos Rh D y RhCE. Objetivo: Determinar la frecuencia de los antígenos C, c, E y e del sistema Rh en donantes de sangre Rh negativos del Hemocentro Centro Oriente Colombiano. Métodos: Estudio descriptivo de corte transversal que incluyó 193 donantes voluntarios de sangre del Hemocentro Centro Oriente Colombiano, la fenotipificación de los antígenos del sistema Rh se realizó utilizando la técnica en tarjeta gel. Se calculó la frecuencia fenotípica de los antígenos del sistema Rh, en porcentajes y para el procesamiento de la información se utilizó el paquete estadístico SPSS versión 21.0 donde se realizó el análisis de los datos de la población. Como criterios de inclusión ser donante voluntario de sangre y de exclusión, estar hemoclasificado como antígeno DEL, D débil y D parcial. Resultados: Las muestras analizadas fueron obtenidas de la tubuladura central de la unidad fraccionada de glóbulos rojos, identificándose tres fenotipos con la siguiente frecuencia: cde/cde (95,86 por ciento), cde/cdE (3,10 por ciento) y cde/Cde (1,04 por ciento). Los participantes del estudio provenían de diversos municipios del departamento de Boyacá y otras regiones del país como llanos Orientales, Santander y Cundinamarca. Los antígenos del sistema Rh son altamente polimórficos a nivel de poblaciones, dada la importancia inmunológica de los antígenos del sistema Rh, los cuales se ven directamente relacionados con el desarrollo de anemia hemolítica postransfusional y perinatal, la fenotipificación ampliada brinda mayor seguridad transfusional y seguimiento al estado del feto o neonato(AU)

Introduction: The Rh system is composed of more than 56 antigens capable of producing hemolytic disease, defined by serological methods; being the most important and therefore the largest antigens of the system, the antigens D, C, c, E and e. These antigens are located on two proteins that are expressed in the membrane of erythrocytes Rh D and RhCE. Objective: To determine the frequency of the C, c, E and e antigens of the Rh system in Rh negative blood donors of Hemocentro Centro Oriente Colombiano. Methods: A cross-sectional descriptive study that included 193 blood donors from the Hemocentro Centro Oriente Colombiano, phenotyping the antigens of the Rh system, using the gel card technique. The phenotypic frequency of the Rh antigen was calculated, in percentages and for the processing of the information, the statistical package SPSS version 21.0 was used in Spanish where all the analysis of the population data was performed. With inclusion criteria to be a voluntary blood donor and exclusion, be hemoclasified as DEL antigen, weak D and partial D. Results: The analyzed samples were obtained from the central tubulant of the fractionated unit of red blood cells, three phenotypes were identified with a frequency of :cde / cde95.86 percent, cde / cdE 3.10 percent and cde / Cde 1, 04 percent; it was evidenced that the participants come from diverse municipalities of the department of Boyacá and other regions of the country like Eastern Plains, Santander and Cundinamarca. The Rh factor and the antigens of the Rh system are highly polymorphic at the population level, given the immunological importance of the antigens of the Rh system, which are directly related to the development of post transfusion hemolytic anemia and perinatal hemolytic disease, the extended phenotyping provides greater transfusional safety and follow-up to the status of the fetus or neonate(AU)

Applicability of papain solutions in immunohematology / Aplicabilidade de soluções de papaína em imuno-hematologia

ABSTRACT Objective: To compare the enzyme activity of different presentations of papain solution to validate in-house preparations. Methods: Two papain solutions were prepared, and the third presentation was a commercial solution. Tests were carried out with samples of red cells typed as weak RhD. Results: In-house prepared papain solutions showed similar enzyme reactivity, and statistically no differences compared to the enzyme activity of the commercial solution. Conclusion: Evaluating the cost-benefit ratio, the in-house prepared papain solutions present more economic advantages, and can be incorporated into immunohematological routines as a way to cope with periods of financial crisis and cost-containment policies.

RESUMO Objetivo: Comparar a atividade enzimática de diferentes apresentações de solução de papaína para validação de preparados in-house. Métodos: Foram preparadas duas soluções de papaína, e a terceira apresentação tratou-se de uma solução comercial. Os testes comparativos das reações enzimáticas foram realizados com amostras de hemácias tipadas como RhD fraco. Resultados: As soluções de papaína preparadas in-house apresentaram reatividade enzimática semelhante e estatisticamente sem diferenças em comparação com a atividade enzimática da solução comercial. Conclusão: Avaliando-se a relação entre custo e benefício, as soluções de papaína preparadas in-house são economicamente vantajosas, podendo ser incorporadas às rotinas imuno-hematológicas como forma de enfrentamento em períodos de crise financeira e em políticas de retenção de gastos.

Effect of 2Me, DTT, ZAAP and Enzyme on JMH Antigen on the Surface of Human Erythrocytes / 中国实验血液学杂志

OBJECTIVE@#To explore the the effects of 2-Me, DTT, papain, pineapple protease and ZZAP on the antigenicity of JMH antigen of human red blood cells (RBC) surface.@*METHODS@#Firstly, human RBC were treated with 2-Me, DTT, pineapple protease, papain and ZZAP reagents, respectively. The antigenicity of JMH antigen on human RBC surface was detected and analyzed by flow cytometry.@*RESULTS@#Flow cytometric analysis found that compared with level before treatment, the antigenicity of JMH antigen on RBC surface was significantly reduced after 2-Me treatment, the positive rate of JMH antigen: 69.5%±4.5% vs 56.5%±3.4% (t=12.44, P＜0.01); fluorescence intensity: 4906±317 vs 3003±165 (t=11.84, P＜0.01). The antigenicity of JMH antigen on RBC surface significantly increased after DTT treatment, showing the positive rate of JMH antigen: 61.7%±3.8% vs 75.5±4.9% (t=16.57, P＜0.01), fluorescence intensity: 4044±294 vs 4854±319 (t=15.46, P＜0.01). However, both bromelain and papain could significantly reduce the antigenicity of JMH antigen on the RBC surface, Bromelain: the positive rate of JMH antigen: 62.2%±3.8% vs 8.8%±1.2% (t=26.44, P＜0.01), fluorescence intensity: 4263±273 vs 1444±212 (t=19.27, P＜0.01); Papain: the positive rate of JMH antigen: 62.8%±3.6% vs 8.8%±1.5% (t=21.38, P＜0.01), fluorescence intensity: 4389±284 vs 1458±230 (t=17.49, P＜0.01). The flow cytometric analysis revealed that ZZAP treatment significantly reduced the antigenicity of JMH antigen on the RBC surface, the positive rate of JMH antigen: 62.2%±4.4% vs 48.2%±4.1% (t=14.87, P＜0.01), fluorescence intensity: 4106±263 vs 2063±175 (t=17.49, P＜0.01).@*CONCLUSION@#The treatment with 2-Me can reduce the antigenicity of JMH antigen on human RBC surface. The antigenicity of JMH antigen on human RBC surface increased after DTT treatment. The antigenicity of JMH antigen on human RBC surface significantly reduces after the treatment with pineapple protease or papain. ZZAP treatment can reduce the antigenicity of JMH antigen on the RBC surface.

Identification of A Case of DEL Blood Group with DAT Positive Phenotype and Pedigree Analysis / 中国实验血液学杂志

OBJECTIVE@#To identify the blood group of a patient with DEL phenotype combined with positive direct anti-human globulin test and to analyze the pedigree.@*METHODS@#Routine serological reagents were used for serological analysis of RhD blood group in the pedigree members. Exons and flanking sequences of RHD gene were amplified, sequenced and analyzed for heterozygosity. The familial genetic state of DEL phenotype was further analyze in the family members.@*RESULTS@#The DAT was strongly positive in the proband. The 1227G>A allele (RHD*DEL1) was present in the exon 9 of RHD gene, and the mother was the carrier of RHD*DEL1. The proband was identified as RHD+/RHD-, suggesting the CDe/Cde haplotype.@*CONCLUSION@#The proband is DEL phenotype (RHD*DEL1).

A case with a novel weak D type / 中华医学遗传学杂志

OBJECTIVE@#To report on a novel weak D type identified in a Chinese individual.@*METHODS@#Peripheral blood sample was collected for a voluntary blood donor with weakened expression of D antigen. Routine serological testing was carried out to determine the D, C, c, E and e antigens of the Rh blood group. A D-screening kit was used to analyze the RhD epitopes. The 10 exons and flanking intronic regions of the RHD gene were sequenced. The zygosity of RHD was determined with a sequence-specific primer PCR method.@*RESULTS@#A novel RHD allele, RHD (1022T>A), was found in the subject with a weak D phenotype. Serological testing of the RhD epitopes has coined with the weak D phenotype.@*CONCLUSION@#A novel weak D allele has been identified in Chinese population.