针对不同基因靶点的胃黏膜标本幽门螺杆菌PCR检测敏感性分析  

Using PCR with different primers to detect Helicobacter pylori in gastric biopsy specimens

作　　者：刘国栋[1] 何利华[1] 顾一心[1] 杨宁敏 张建中[1] 

机构地区：[1]中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室,北京102206 [2]杭州致远医学检验中心

出　　处：《中国病原生物学杂志》2011年第6期401-403,410,共4页Journal of Pathogen Biology

基　　金：国家"十一五"科技支撑计划项目(No.2007BAD4B02)

摘　　要：目的分析针对3种不同基因靶点的胃黏膜标本幽门螺杆菌PCR检测的敏感性,评价PCR方法从胃黏膜标本中检测幽门螺杆菌用于临床诊断的价值。方法采集320例13C呼气试验阳性患者的胃黏膜标本,用针对ureA、ca-gA和vacA基因的3对引物进行PCR检测,同时进行幽门螺杆菌分离培养。分别对3对引物的PCR结果相互进行Kappa一致性检验,并比较组合PCR与培养结果。结果幽门螺杆菌培养的阳性率为81.88%(262/320);ureA、cagA和vacA PCR扩增阳性率分别为56.25%(180/320)、58.13%(186/320)和81.56%(261/320),组合PCR阳性率为90.94%。ureA、cagA和vacA PCR结果经一致性检验,Kappa值分别为0.209、0.137、0.129,检测结果具有互补性。结论单一基因PCR扩增的敏感性较低且不同基因扩增效果间的一致性较差。3对引物组合使用可提高PCR方法的敏感性,幽门螺杆菌检出率高于培养法,具有一定的实际应用价值。Objective The aim of this study was to assess the sensitivity of three primers and the feasibility of using PCR to detect Helicobacter pylori in gastric biopsy specimens.Methods Gastric biopsy specimens were collected from 320 patients who tested positive on a 13C urea breath test.H.pylori was detected by culturing and PCR with three primers（ureA,cagA,and vacA）.The concordance between PCR results with the three primers was accessed using a kappa concordance test,and the difference between culturing and PCR results was also assessed.Results Cultures were positive for H.pylori at a rate of 81.88%（261/320）.PCR with the primers ureA,cagA,and vacA was positive for H.pylori at a rate of 56.3%（180/320）,58.1%（186/320）,and 81.6%（262/320）,respectively,and combined PCR was positive for H.pylori at a rate of 90.9%（291/320）.There concordance between PCR results with the three primers,but that concordance was slight.Kappa values were 0.209,0.137,and 0.129,respectively.PCR results with different primers complemented one another.Conclusion PCR using a single primer had a low sensitivity,and there was little concordance between results with each primer.Using multiple primers can improve the sensitivity of PCR detection and might help to detect H.pylori in gastric biopsy specimens.

关 键 词：PCR 胃黏膜标本 幽门螺杆菌 培养 

分 类 号：R378.99[医药卫生—病原生物学]