摘要
目的:建立基于实时荧光定量PCR(qPCR)技术的Diego、MNS、Kell血型基因分型方法,并初步应用于献血者稀有血型筛查。方法:利用血型血清学方法和基于测序分型的PCR-SBT法,制备包含各等位基因杂合子和纯合子的血型基因标准品。设计血型基因特异性扩增引物和杂交探针,摸索建立qPCR检测Diego、MNS、Kell血型基因型的方法。利用建立的qPCR法对186例献血者进行血型鉴定。结果:建立了基于qPCR技术鉴定Dia/Dib、S/s、K/k血型抗原的方法,基因标准品检测分型结果与血清型及PCR-SBT检测的基因型完全一致。采用qPCR法检测186份献血者样本,发现11例DI A/B杂合型,19例GYPB S/s杂合型,其余均为DI B/B、GYPB s/s、KEL 02/02纯合型,未发现DI A/A、GYPB S/S、KEL 01.01/01.01稀有血型。结论:建立的qPCR方法适用于Diego、MNS、Kell血型基因分型,可用于献血者批量筛查和稀有血型建库。
Objective:To establish a genotyping method for Diego,MNS and Kell blood groups based on quantitative real-time PCR(qPCR)technology,and preliminarily apply it to the screening of rare blood groups in blood donors.Methods:Blood group gene standards containing heterozygous and homozygous alleles were prepared by blood group serological and PCR-SBT methods.Specific amplification primers and hybridization probes were designed,and explore to establish the qPCR method for detecting Diego,MNS,and Kell blood group genotypes.Then the established qPCR method was used to identify blood group genotypes of 186 blood donor samples.Results:A method based on qPCR technology was established to identify Dia/Dib,S/s and K/k blood group antigens.The genotyping results of the gene standard samples were consistent with the serological testing results and genotypes detected by PCR-SBT.qPCR testing of 186 samples identified 11 cases of DI A/B heterozygosity and 19 cases of GYPB S/s heterozygosity,and the rest were DI B/B,GYPB s/s,KEL 02/02 homozygosity.No rare blood group genotypes of DI A/A,GYPB S/S,KEL 01.01/01.01 were found.Conclusion:The established qPCR method is suitable for genotyping on Diego,MNS and Kell blood group,and it can be used for batch screening of blood donors and the establishment of rare blood group bank.
作者
张兵
徐罡
胡文健
洪小珍
许先国
ZHANG Bing;XU Gang;HU Wen-Jian;HONG Xiao-Zhen;XU Xian-Guo(Institute of Transfusion Medicine,Blood Center of Zhejiang Province,Hangzhou 310052,Zhejiang Province,China;Yiwu Central Blood Station,Jinhua 322000,Zhejiang Province,China)
出处
《中国实验血液学杂志》
北大核心
2025年第5期1429-1434,共6页
Journal of Experimental Hematology
基金
浙江省血液安全研究重点实验室开放课题(2021 KF003)。
作者简介
通信作者:许先国,主任技师.E-mail:yjs001@163.com。
