OmpA-VS2为靶基因的非标记实时荧光PCR检测沙眼衣原体感染  

A non-labeled real-time PCR assay targeting OmpA-VS2 for detection of Chlamydia trachomatis

作　　者：李建红[1] 尹跃平[1] 钟铭英[1] 彭锐锐[1] 郑和平[2] 陈祥生[1] 

机构地区：[1]中国医学科学院北京协和医学院皮肤病研究所中国疾病预防控制中心性病控制中心,江苏南京210042 [2]广东省皮肤性病防治中心,广州510500

出　　处：《中国艾滋病性病》2010年第2期109-112,共4页Chinese Journal of Aids & STD

基　　金：联合国开发计划署/世界银行/世界卫生组织热带病研究与培训特别规划署(TDR)项目(项目编号A70578)~~

摘　　要：目的建立一种以OmpA-VS2为靶基因的非标记实时荧光聚合酶链反应(PCR)方法,以检测沙眼衣原体。方法设计15种型别沙眼衣原体通用引物,对OmpA-VS2进行套式PCR扩增和实时荧光PCR检测,并对其敏感性、特异性进行评价,并用临床标本进行验证。结果15种型别沙眼衣原体均扩增出168bp的目的片段;敏感性为每个反应1个拷贝。特异性分析可见,与10种泌尿生殖道病原体和共生菌均没有交叉反应。临床标本验证的结果显示,该方法与以质粒为靶基因的核酸扩增检测的Amplicor CT/NG方法的检测结果一致。结论该方法具有敏感性高、特异性强、扩增后无需PCR后处理等特点,可望用于临床与防治项目中沙眼衣原体的检测。Objective To establish a non-labeled real-time PCR assay targeting OmpA-VS2 for detection of Chlamydia trachomatis （C. trachomatis）. Methods Two pairs of universal primers were designed to amplify the OmpA-VS2 region of fifteen genotypes of C. trachomatis by the nested real-time PCR. The sensitivity and specificity of the assay were analyzed and further validated with clinical positive specimens. Results All 15 genotypes of C. trachomatis were successfully amplified with a product of 168bp. The detection limit with the assay was 1 copy per reaction mixture and no cross reactions with 10 other urogenital pathogens or commensals were found. The clinical validation reveals a coincidence with the results detected by plasmid PCR. Conclusion The assay is characterized with high sensitivity and specificity, and no separated post-PCR analysis is needed. It would be an appropriate assay to be used for detection of chlamydial infection in clinical practice as well as in disease control programs.

关 键 词：沙眼衣原体 OmpA基因 实时PCR 

分 类 号：R374.1[医药卫生—病原生物学]