山东临沂地区RhD阴性无偿献血人群中RHD基因变异体的分析

Analysis on RHD variants among RhD negative voluntary blood donors in Linyi district of Shandong Province

目的探讨临沂地区RhD阴性无偿献血人群RHD基因的变异体及其分子背景。方法 2011年采集24 875人份全血,获得RhD初筛阴性标本111例,提取DNA后,采用序列特异性引物-多聚酶链反应(SSP-PCR),进行以下三步实验:①检测出缺失RHD基因的RhD阴性、DEL1227G>A突变以及其他变异体;②鉴定其他变异体的具体类型;③对有突变位点未检出型别的变异体进行测序。结果临沂地区无偿献血人群中RhD阴性比例为0.45%,在111例RhD初筛阴性标本中,检测到1～10外显子全缺失76例,2～9外显子缺失9例,845G>A突变3例,3～6外显子缺失3例,1227G>A突变18例,RHD(711DELC)1例,Dva(Hus)1例。结论临沂地区常规血清学筛查RhD阴性无偿献血人群中RHD基因存在多态性。

Objective To elucidate the molecular background of RHD gene among RhD negative voluntary blood donors in Linyi district of Shandong province.Methods A total of 24 875 blood samples were collected in 2011.Genomic DNA from 111 RhD negative samples were analyzed by Sequence specific primer-polymerase chain reaction（SSP-PCR） for three steps： ① to detect RHD-deleted RhD negative samples,DEL1227GA mutations and other variants;② to identify the gene type of other variants;and ③ to perform sequencing procedure for the variants which had mutation sites but the gene type was unable to be identified.Results The proportion of RhD negative blood donors was 0.45% among voluntary blood donation population in Linyi district.In 111 RhD negative samples,there were 76 with deletion of 1-10 exons,9 with deletion of 2-9 exons,3 with 845GA mutation,3 with deletion of 3-6 exons,18 with 1227GA mutation,and 1 with RHD（711DELC）,1 sample with DVa（Hus）.Conclusion There are RHD polymorphisms among RhD negative voluntary blood donors in Linyi district.