基于双Taqman探针实时荧光PCR技术的HLA-B*5801基因检测方法建立与应用  

Development and application of a method for HLA-B~*5801 detection based on double Taqman PCR

作　　者：赵学峰 袁晓文 谭荧飞 罗丽贞 ZHAO Xue-feng;YUAN Xiao-wen;TAN Ying-fei;LUO Li-zhen(Laboratory Department of Affiliated Nanhai Hospital of Southern Medical University,Foshan,Guangdong 528200,China)

机构地区：[1]南方医科大学附属南海医院检验科,广东佛山528200

出　　处：《热带医学杂志》2020年第11期1402-1404,1437,共4页Journal of Tropical Medicine

基　　金：佛山市科技局课题(20180158)。

摘　　要：目的建立一种基于Taqman探针实时荧光PCR技术的HLA-B*5801基因检测方法;调查佛山地区HLA-B*5801基因阳性携带率。方法根据HLA-B*5801等位基因外显子2和3的基因序列设计引物和Taqman探针,建立一种实时荧光PCR检测方法,与商业试剂盒和Sanger测序进行对比检测性能评价。选取健康体检男女样本各200例,采用本方法检测并调查分析本地区汉族人群HLA-B*5801基因携带率。结果本研究建立的HLA-B*5801实时荧光PCR检测方法与2种商业试剂盒和Sanger测序的检测结果相符性为100%;佛山地区汉族人群男女HLA-B*5801基因携带率分别为8.5%和11.5%,差异无统计学意义(P=0.48)。结论本研究建立的双Taqman探针HLA-B*5801基因检测方法操作简单、结果准确,可应用于常规检测。Objective To establish a method for HLA-B~*5801 gene detection based on double Taqman real-time fluorescence PCR,and investigate the frequency of HLA-B~*5801 in Han population of Foshan.Methods Primers and Taqman probes were designed according to the exon 2 and exon 3 sequences of HLA-B~*5801 gene,subsequently a method was developed.It was evaluated by comparing with the commercial kits and Sanger sequencing.The radom selected 200 males and 200 females from physical examination were detected with this method to investige the HLA-B~*5801 gene carrying rate in Han population of this region.Results The consistence between this method and the commercial kits/Sanger sequencing were 100%.The HLA-B~*5801 gene carrying rate was 8.5% and 11.5% of male and female,respectively in Foshan Han population,which had no significant difference between male and female(P=0.48).Conclusion This double Taqman probes based method for HLA-B~*5801 gene detection is simple and accurate,which can be applied in routine detection.

关 键 词：HLA-B*5801 TAQMAN探针 实时荧光PCR 

分 类 号：R446[医药卫生—诊断学]