摘要
Objectives: To develop a multi-nested polymerase chain reaction in an assay to detect early Treponema pallidum and Haemophilius ducreyi DNA in the swabs of genital ulcers. Methods: Four pairs of outer and inner primers, specific to the basic membrane protein gene of Treponema pallidum and to the 16s rRNA gene of H ducreyi were synthesized. The multi-nested PCR was developed and applied to detect Treponema pallidum and Haemophilus dicreyi in clinical swabs. Result: The two samples of standard strains of Haemophilus ducreyi and one Treponema pallidum were amplified and showed 309-bp rRNA gene of Haemophilus ducreyi and 506-bp DNA of Treponema palidum, respectively. Out of 51 samples of genital ulcer detected, 29 showed Treponema pallidum positive product and no Haemophilus ducreyi DNA was found. Conclusion: The multi-nested PCR for Treponema pallidum and Haemophilus ducreyi could be useful for early detection and distinguishing diagnosis between syphilis and chancroid.
目的为了建立一种多重巢式聚合酶链反应方法用于快速检测生殖器溃疡拭子中梅毒螺旋体和杜克雷嗜血杆菌。方法根据梅毒螺旋体基础膜蛋白基因和杜克雷嗜血杆菌的16sRNA基因分别设计和合成2对梅毒螺旋体特异性引物和2对杜克雷嗜血杆菌特异性引物,建立多重巢式聚合酶链反应,同时检测临床生殖器溃疡拭子标本中的梅毒螺旋体和杜克雷嗜血杆菌。结果1株梅毒螺旋体标准株和2株杜克雷嗜血杆菌标准株分别扩增出506-bp和:309-bp基因片断。检测51例临床标本,梅毒螺旋体阳性29例,杜克雷嗜血杆菌均阴性。结论多重巢式聚合酶链反应可用于梅毒螺旋体和杜克雷嗜血杆菌的早期检测和鉴别诊断。
