重庆地区RhD变异型无偿献血者的基因多态性及其表型研究  

The gene polymorphism and phenotype of RhD variants among blood donors in Chongqing

作　　者：刘静怡 崔丹荔 王芳[1] 黎美君 刘东[1] 谢晓艳 陈敏 付威义 杨冬燕[1] 张巧琳[1] LIU Jingyi;CUI Danli;WANG Fang;LI Meijun;LIU Dong;XIE Xiaoyan;CHEN Min;FU Weiyi;YANG Dongyan;ZHANG Qiaolin(Chongqing Blood Center,Chongqing 400015,China;Xi′an Haorui Genomics Technology Company Limited)

机构地区：[1]重庆市血液中心,重庆400015 [2]西安浩瑞基因技术有限公司

出　　处：《中国输血杂志》2024年第8期879-885,共7页Chinese Journal of Blood Transfusion

基　　金：重庆市科卫联合医学科研项目(2023QNXM042);重庆市九龙坡区科技计划项目(2022-02-018-Y);重庆市卫生健康委医学科研项目(2024WSJK033)。

摘　　要：目的对重庆地区22例RhD变异型无偿献血者标本进行Rh血型血清学检测和三代基因测序,了解重庆地区RhD变异型的表型分布及其基因分型。方法选择2023年1—8月本中心参与无偿献血的人群作为研究对象。使用传统血清学方法对其进行RhD表型鉴定,确定为RhD变异型后使用D-screen试剂盒对其进行RhD不同抗原表位的检测。此外,提取其基因组DNA,使用叠瓦式引物设计进行多段扩增、拼接获得RHD基因全长序列进行三代测序检测,并用SnapGene软件对序列结果进行分析。结果在22例RhD变异型中,8例为DVI 3型(36.36%),其存在RHD-CE(3-6)-D杂交等位基因;6例部分弱D15型(27.27%),主要发生的突变为c.845G>A;6例亚洲型Del(27.27%),主要发生的突变为c.1227G>A,还有1例弱D17型发生的突变为c.340C>T和1例推测为部分D(c.491A>T,p.Asp164Val,错义突变)。结论重庆地区无偿献血人群中最常见的RhD变异型为DVI 3型,使用SMRT三代测序技术可以获得RhD变异型单倍体全长。Objective To conduct Rh blood group serological testing and third-generation sequencing(TGS)on 22 RhD variant voluntary blood donors in Chongqing and explore the phenotypic distribution and genotyping of RhD variants in Chongqing.Methods From January to August 2023,individuals who participated in blood donation in our blood center were selected as the study objects.RhD variant phenotype identification was performed using routine serological methods.Once the RhD variants were identified,tests on different antigenic epitopes of RhD were conducted using a D-screen assay kit.Furthermore,after the genomic DNA from 22 RhD variant blood samples was extracted,imbraided primers design and multi-segment amplification and splicing were used to sequence the full-length RHD gene for TGS.The RHD gene sequence was analyzed using SnapGene software.Results Among the 22 RhD variants,8 were DVI type 3(36.36%),with the main mutation of RHD-CE(3-6)-D hybrid allele.Six cases(27.27%)showed partial weak D15 type,with the main mutation of c.845G>A.There were 6 cases of Asia type Del(27.27%),with the main mutation of c.1227G>A.One case was weak D17 type with a mutation of c.340C>T and 1 case speculated to be partial D(c.491A>T,p.Asp164Val,missense mutation).Conclusion The most common RhD variant phenotype among blood donors in Chongqing is DVI type 3,and the full-length haplotype sequence of RHD variant alleles can be obtained by Pacific Bioscience single-molecule real-time sequencing(SMRT).

关 键 词：RhD变异型 RHD基因 SMRT 三代测序技术 血清学 重庆 

分 类 号：Q343.13[生物学—遗传学] R457.11[医药卫生—治疗学] S852.43[医药卫生—临床医学]