性病病原体多重聚合酶链反应检测方法的建立和验证

Multiplex polymerase chain reaction assay for simultaneous detection of sexually transmitted pathogens

目的建立同时检测淋球菌、沙眼衣原体、解脲支原体和单纯疱疹病毒-2致病微生物的多重聚合酶链反应(PCR)方法,用于四种性病的诊断和治疗监测。方法根据淋球菌外膜孔蛋白基因、沙眼衣原体基因、解脲支原体尿素酶基因和单纯疱疹病毒-2的DNA聚合酶基因序列,各设计一对特异性引物,优化反应体系和条件进行多重PCR反应。结果 4对引物分别扩增出327、167、426、391bp的目的条带,并且特异性强,与其他非目的病原体基因不发生反应。结论本研究建立的多重PCR方法为相关病原体的快速检测提供方法。

Objective To develop a multiplex polymerase chain reaction （PCR） method for simuhaneous detection of sexually transmitted pathogens. Methods According to specific gene sequence of four pathogens： porin protein of neisseria gonorrhoeae, chlamydia trachomatis, urease of ureaplasma urealyticum and DNA polymerase of human herpesvirus 2, four sets of specific primers were designed and a multiplex PCR assay was developed to detect 4 pathogens in one test based on the optimized reaction system and condition. Results The target DNA at 327, 167, 426 and 391 bp of 4 pathogens were specifically amplified by their specific primers. The DNA of other several common pathogens including gardnerella vaginalis, trichomoniasis, acinetobacter baumannii was not amplified by four sets of primers. Conclusion It was shown that a simultaneous, specific and rapid multiplex PCR assay method was successfully constructed to detect four sexually transmitted pathogens.