应用分子生物学测序技术对血小板抗原HPA-3进行基因分型  

Genotyping of human platelet antigens of HPA-3 system by PCR-SBT

作　　者：廖扬勋 陈志忠 丁浩强[2] 余文潮 梁洁贞 陈超红 李结敏 LIAO Yang-xun;CHEN Zhi-zhong;DING Hao-qiang;YU Wen-chao;LIANG Jie-zhen;CHEN Chao-hong;LI Jie-min(Zhaoqing Blood Center, Zhaoqing , Guangdong 526040;Guangzhou Blood Center, Guangzhou , Guangdong 510095, China)

机构地区：[1]肇庆市中心血站,广东肇庆526040 [2]广州血液中心,广东广州510095

出　　处：《热带医学杂志》2017年第12期1597-1600,共4页Journal of Tropical Medicine

基　　金：广东省肇庆市科技计划项目(2014F006)

摘　　要：目的利用分子生物学测序技术聚合酶链反应直接测序方法(PCR-SBT))对血小板抗原(HPA)-3系统进行基因分型。方法采用本研究合成的引物,应用分子生物学测序技术对101名随机血小板捐献者进行HPA-3基因分型,随机抽样20个标本进行聚合酶链反应-序列特异性引物(PCR-SSP)基因分型作为平行对照,质控DNA进行验证。结果最终选取61℃的为理想退火温度。采用合成的引物对4份质控标本作HPA-3基因分型:与Innotrain提供的结果完全一致。101名随机汉族血小板志愿捐献者HPA基因频率为HPA-3a 0.520、HPA-3b 0.480;随机抽样20个样品与Inno-train公司的HPA-SSP分型结果一致。结论所建立的HPA基因PCR-SBT分型技术分辨率高,正反向引物测序结果一致,都可以精确分型,可以弥补PCR-SSP不足。Objective Molecular biology sequencing technology was used for genotyping of human platelet antigen-3 （HPA- 3 ） system. Methods Primers were designed for the genotyping of HPA-3 system. 101 platelet samples were collected from the donors and the HPA- 3 system genotypes were determined by polymerase chain reaction-sequence based typing （PCR- SBT）. 20 samples were genotyped by PCR-SSP for comparison. Results 61℃ was determined as the optimal annealing temperature for PCR-SBT. The results of PCR-SBT and PCR-SSP were consistent. The genotype frequencies of HPA-3a and HPA-3b were 0.520 and 0.480, respectively. Conclusions PCR-SBT assay established in our study provides a high resolution and consistent results. The assay can accurately determine the genotype of HPA- 3 system, and can make up the deficiency of PCR-SSP.

关 键 词：人类血小板抗原 HPA-3系统 PCR-SBT 基因分型 

分 类 号：Q503[生物学—生物化学]