新乡地区Rh阴性无偿献血者Rh表型分布及RhD变异型基因型分析  

Analysis of Rh phenotype distribution of Rh negative unpaid blood donors and RhD variant genotype in Xinxiang area

作　　者：赵甜 张晨光 庞桂芝 ZHAO Tian;ZHANG Chenguang;PANG Guizhi(Department of Clinical Laboratory,Xinxiang Central Hospital,Xinxiang 453000,Henan Province,China;School of Public Health,Xinxiang Medical University,Xinxiang 453003,Henan Province,China;Blood Group Room of Xinxiang Central Blood Station,Xinxiang 453001,Henan Province,China)

机构地区：[1]新乡市中心医院检验科,河南新乡453000 [2]新乡医学院公共卫生学院,河南新乡453003 [3]新乡市中心血站血型室,河南新乡453001

出　　处：《新乡医学院学报》2023年第5期458-461,共4页Journal of Xinxiang Medical University

摘　　要：目的分析新乡地区Rh阴性无偿献血者Rh表型分布及RhD变异型基因型。方法选择2018年1月至2019年12月于新乡市中心血站无偿献血的110667名无偿献血者为研究对象。采用试管法ABO正反定型试验检测ABO血型,采用试管法Rh抗原分型试验检测Rh表型并初筛RhD阴性受试者,采用间接抗人球蛋白试验对初筛RhD阴性受试者进行RhD阴性确认,并检出RhD变异型受试者。采用聚合酶链式反应法扩增RhD变异型受试者RHD基因外显子E1~E10,将扩增产物进行琼脂糖凝胶电泳,观察相应的外显子区域有无特异性条带出现。采用Sanger测序法对RhD变异型受试者的RHD基因外显子E1~E10的特异性扩增产物进行基因测序,应用SeqMan软件分析基因序列,根据单核苷酸多态性位点确定RhD变异型。结果110667名受试者中,初筛RhD阴性269例(0.24%)。269例初筛RhD阴性受试者中,Rh阴性256例(95.17%),RhD变异型13例(4.83%)。256例RhD阴性受试者中,ABO血型为A型78例(30.47%)、B型92例(35.94%)、O型64例(25.00%)、AB型22例(8.59%),Rh表型分布以ccdee(60.55%)和Ccdee(29.30%)居多。11例RhD变异型受试者的RHD基因外显子E1~E10为特异性条带,2例RhD变异型受试者的RHD基因外显子E1~E5、E8、E10为特异性条带。13例RhD变异型受试者中,7例弱D型、4例DEL型和2例部分D型。13例RhD变异型受试者RHD基因RhCE表型分别为CCee(2/13)、Ccee(6/13)和ccEe(5/13)。7例RhD变异型受试者等位基因RHD*15的E6外显子发生845G>A杂合突变。4例RhD变异型受试者的E9外显子发生1227 G>A杂合突变,其中1例受试者同时发生ZVS7+152 C>A突变。2例RhD变异型受试者的RHD基因E6~E9外显子发生突变,被RHCE基因所取代,表现为RHD-CE(6-9)-RHD。结论新乡地区RhD阴性无偿献血者中Rh表型分布以ccdee和Ccdee居多,RhD变异型以弱D型为主,其RHD等位基因为RHD*15。Objective To analyze the Rh phenotype distribution and RhD variant genotype of Rh negative unpaid blood donors in Xinxiang area.Methods A total of 110667 voluntary blood donors who donated blood at the Xinxiang Central Blood Station from January 2018 to December 2019 were selected as the research subjects.ABO blood type was detected by using the ABO positive and negative stereotyping test by tube method,Rh phenotype was detected by the in vitro Rh antigen typing test,and RhD negative subjects were initially screened.The RhD negative subjects in the initial screening were confirmed by indirect anti human globulin test,and RhD variant subjects were ultimately detected.The exons E1-E10 of RHD gene of RhD variant subjects were amplified by polymerase chain reaction,and the amplified products were subjected to agarose gel electrophoresis to observe whether there were specific bands in the corresponding exon regions.The specific amplification products of RHD gene exons E1-E10 in RhD variant subjects were sequenced by the Sanger sequencing method,the gene sequence was analyzed by SeqMan software,and the types of RhD variants was determined based on single nucleotide polymorphism loci.Results Among 110667 subjects,269 cases(0.24%)were negative for RhD during initial screening.Among the 269 initially screened RhD negative subjects,256 cases were RhD negative(95.17%)and 13 cases were RhD variant(4.83%).Among the 256 RhD negative subjects,there were 78 cases(30.47%)of type A,92 cases(35.94%)of type B,64 cases(25.00%)of type O,and 22 cases(8.59%)of type AB.The Rh phenotype distribution was mainly ccdee(60.55%)and Ccdee(29.30%).Eleven RhD variant subjects showed specific bands in exons E1-E10 of the RHD gene;two RhD variant subjects showed specific bands in exons E1-E5,E8,and E10 of the RHD gene.Among the 13 RhD variant subjects,7 cases were weak D type,4 cases were DEL type,and 2 cases were partial D type.The RhCE phenotypes of the RHD gene in 13 RhD variant subjects were CCee(2/13),Ccee(6/13),and ccEe(5/13),respectively.Sev

关 键 词：D变异型 RHD基因 Sanger测序法 输血治疗 

分 类 号：R457.11[医药卫生—治疗学]