荧光定量聚合酶链反应染料法在HLA-B~★5801等位基因分析中的应用

Application of real-time quantitative polymerase chain reaction in analyzing HLA-B*5801 allele

目的:评估荧光定量聚合酶链反应(PCR)染料法在HLA-B*5801等位基因分析中的灵敏度、特异度、准确度和Kappa值,并与经典测序方法进行比较。方法:应用HLA-B*5801基因检测试剂盒对268例痛风、痛风性关节炎和高尿酸血症患者和50例正常人HLA-B*5801进行分析,并对318例标本HLA-B基因2、3、4外显子双向测序。结果:荧光定量PCR染料法敏感度、特异度和准确度均为100%,Kappa值为1。318例参与者HLA-B*5801等位基因发生率为9.2%(29/318)。痛风患者HLA-B*5801发生率8.51%(8/94),痛风性关节炎发生率9.52%(2/21),高尿酸血症发生率11.11%(17/153),正常人发生率4%(2/50)。结论:荧光定量PCR染料法能够快速准确检测HLA-B*5801,检测结果与测序方法完全一致。

Objective：To evaluate the sensitivity,specificity,accuracy and Kappa value of real-time quantitative polymerase chain reaction （PCR）with fluorescent dye in detecting HLA-B＊5801allele,and compare the results with the sequence based typing （SBT）.Methods：A total of 268 goat,goaty arthritis and hyperuricemia patients and 50 normal control were analyzed using HLA-B＊5801 gene kit,and both directions of exons 2,3,4 of HLA-B gene in 318 cases were sequenced.Results：The sensitivity,specificity and accuracy of real-time quantitative PCR with fluorescent dye were all 100％,and the Kappa value was 1.The frequency of HLA-B＊5801in 318 participants was 9.2％（29/318）,and the frequencies of HLA-B＊5801 in goat,goaty arthritis and hyperuricemia were 8.51％（8/94）,9.52％（2/21）,11.11％（17/153）,alternatively,and the normal control was 4％（2/50）.Conclusion：The method of real-time quantitative PCR with fluorescent dye can detect HLA-B＊5801 rapidly and precisely,and the results are consistent with the results of SBT.