荧光定量PCR染料法在HLA-B27检测中的应用  

Evaluation of fluorescence quantitative PCR method on detecting HLA-B27 gene

作　　者：富显果 唐宝佳 杨菁 曹罗元 叶作东 黄宝英 FU Xian-guo;TANG Bao-jia;YANG Jing(Central Laboratory, Ningde Hospital, Fujian Medical University;Department of Hematology and Rheumatology , Ningde Hospital, Fujian Medical University;Department of Clinical Laboratory, Ningde Hospital,Fujian Medical University ,Ningde 352100 ,China)

机构地区：[1]福建医科大学附属宁德市医院中心实验室 [2]福建医科大学附属宁德市医院检验科,福建宁德352100 [3]福建医科大学附属宁德市医院血液风湿科

出　　处：《中国实验诊断学》2018年第6期939-942,共4页Chinese Journal of Laboratory Diagnosis

基　　金：福建省宁德市科技计划资助项目(20100133)

摘　　要：目的探讨荧光定量PCR染料法检测HLA-B27基因对于诊断强直性脊柱炎的临床应用价值。方法应用流式细胞术和荧光定量PCR技术对强直性脊柱炎疑似患者170例进行HLA-B27基因及其亚型检测,对结果不一致样本,进一步应用PCR-直接测序法进行B27测序分析。结果两种方法检测符合率为92.9%,差异无统计学意义(P>0.05);5例结果不一致样本为流式细胞分析阳性,而荧光定量PCR法和PCR-直接测序分析为B*40亚型或B*40与B*58的杂合型;在158例AS相关患者中,共检出3种HLA-B27等位基因:B*2704为主要亚型,占96.8%,其次为B*2705,占1.9%,另有2例为B*2706,占1.3%,结论荧光定量PCR染料法可有效提高HLAB27检测的准确性,并能够实现HLA-B27亚型分析,可为深入探讨HLA-B27在AS中发病中的作用提供依据。Objective To investigate the clinical value of fluorescence quantitative PCR method on detecting HLAB27 gene in the diagnosis of ankylosing spondylitis.Methods HLA-B27 gene and its subtype of 170 patients with suspected ankylosing spondylitis were detected using flow cytometry and fluorescence quantitative PCR.The inconsistent samples were further analyzed by PCR-direct sequencing.Results The coincidence rate of two methods was 92.9%（P〈0.05）.Five cases positive for flow cytometry but negative for quantitative PCR,were identified as B＊40 or heterozygous allele of B＊40 and B＊58 by PCR-direct sequencing.Five B27 alleles were identified among 158 AS related patients：B＊2704,B＊2705,B＊2706,B＊2707.The frequencies for these three alleles were 96.8,1.9 and 1.3 respectively.Conclusion The quantitative PCR method can effectively improve the accuracy of HLA-B27 detection and analyze the alleles of HLA-B27,which then provide evidence for further study on HLA-B27 in the pathogenesis of AS.

关 键 词：等位基因特异性PCR 荧光定量PCR 流式细胞术 强直性脊柱炎 HLA-B27 

分 类 号：R593.23[医药卫生—内科学]