Genotyping analysis of MNS blood group GP(B-A-B) hybrid glycophorins in the Chinese Southern Han population using a high-resolution melting assay
Genotyping analysis of MNS blood group GP(B-A-B) hybrid glycophorins in the Chinese Southern Han population using a high-resolution melting assay
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DOI：
10.1111/trf.14641
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发表时间：
2018-07-01
期刊：
影响因子：
2.9
通讯作者：
Ji, Yanli
中科院分区：
文献类型：
--
作者：
Wei, Ling;Lopez, Genghis H.;Ji, Yanli
mns杂交GP（B-A-B）糖蛋白更常见于东南亚，其携带抗原的同种抗体具有临床意义。由于缺乏商用试剂，血清学技术对杂交糖蛋白的检测是有限的。本研究采用基于高分辨率熔融（HRM）分析的GP（B-A-B）杂交糖蛋白基因分型方法对中国南方汉族人群进行基因分型分析。研究设计与方法采集3104例南方汉族献血者的dna样本。HRM法分析GYP（B-A-B）基因型。部分样品（n=106）也通过多重连接依赖探针扩增（MLPA）检测。对不同熔点曲线的样品进行直接测序。结果共鉴定出5个GYP（B-A-B）基因型（201/3104,6.5%），分别为GYP*Mur杂合子（n=194）、GYP*Mur纯合子（n=3）、GYP*Bun杂合子（n=2）、GYP*HF杂合子（n=1）和一个新的GYP（B-A-B）杂交等位基因（n=1）。HRM对GYP*Mur和野生型GYPB样品的基因分型结果与MLPA一致，而HRM鉴定的GYP*Bun和GYP*HF杂合子，MLPA只能鉴定出GYPB伪外显子3的5个无活性剪接位点1个拷贝。此外，在31份样品中鉴定出10个单核苷酸多态性（SNPs），其中包括4个已知snp和6个新snp。其中一个样本同时携带GYP*Mur和GYP*Sch等位基因。结论HRM法能较好地鉴别出GYP（B-A-B）杂交等位基因。在为中国人群开发GYP（B-A-B）基因分型试剂盒时，应考虑到在GYPB基因内发现的多态性。
BACKGROUNDMNS hybrid GP(B-A-B) glycophorins are more commonly found in Southeast Asians and alloantibodies to antigens they carry are clinically significant. Detection of hybrid glycophorins by serologic techniques is limited due to lack of commercial reagents. In this study, a genotyping method for GP(B-A-B) hybrid glycophorins based on high-resolution melting (HRM) analysis was applied for genotyping analysis in the Chinese Southern Han population.STUDY DESIGN AND METHODSDNA samples from 3104 Chinese Southern Han blood donors were collected. GYP(B-A-B) genotypes were analyzed by HRM assay. Parts of samples (n=106) were also tested by multiplex ligation-dependent probe amplification (MLPA) assay. Direct sequencing was conducted in samples with variant melting curve profiles.RESULTSA total of five GYP(B-A-B) genotypes (201/3104, 6.5%) were identified, which were GYP*Mur heterozygote (n=194), GYP*Mur homozygote (n=3), GYP*Bun heterozygote (n=2), GYP*HF heterozygote (n=1), and a novel GYP(B-A-B) hybrid allele (n=1). Genotyping results for GYP*Mur and wild-type GYPB samples obtained by HRM were consistent with MLPA, while GYP*Bun and GYP*HF heterozygote identified by HRM could only be identified to have one copy of 5 inactive splice site of GYPB Pseudoexon 3 by MLPA. In addition, 10 single-nucleotide polymorphisms (SNPs) including four known and six novel SNPs were identified in 31 samples. One sample was identified carrying both GYP*Mur and GYP*Sch alleles.CONCLUSIONThe HRM assay could distinguish the GYP(B-A-B) hybrid alleles successfully. Polymorphisms identified within the GYPB gene should be taken into consideration when developing GYP(B-A-B) genotyping kits for the Chinese population.