抗原捕获聚合酶链反应分型检测妇女生殖器单纯疱疹病毒

目的 :　建立直接分型检测妇女生殖器单纯疱疹病毒 (HSV)的抗原捕获聚合酶链反应 (AC -PCR)。方法 :　用抗HSV型共同性糖蛋白单克隆抗体 ,包被聚苯乙烯离心管 ,捕获HSV ,同时加入 3个引物 :HSV - 1 HSV - 2型共同性上游引物及HSV - 1和HSV - 2型特异性下游引物 ,进行PCR扩增。结果 :HSV - 1和HSV - 2标准病毒株均分别扩增出与设计大小相符的 4 77bp和 399bpDNA条带。AC -PCR可检测到 10PFUHSV - 1和 1PFUHSV - 2。用AC -PCR检测了 36 5份妇女生殖器拭子标本 ,阳性 10 1例(2 7.7% ) ,2 3例为HSV - 1(占 2 2 .8% ) ,78例为HSV - 2 (占 77.2 % ) ;其中 112份标本同时用AC -PCR和分离培养法检测 ,AC -PCR的阳性率为 2 6 .8% (30 112 ) ,分离培养法的阳性率为 2 0 .5 % (2 3 112 ) ,两者差异有显著性 (χ2 =4 .5 ,P <0 .0 5 )。结论 :　AC -PCR是特异、敏感、快速分型检测妇女生殖器HSV感染的方法

Detection and typing of herpes simplex virus DNA in genital specimens from female patients by antigen capture polymerase chain reaction

Objective: To develop a method for the detection and direct typing of herpes simplex virus (HSV) DNA in genital specimens from female patients by antigen capture polymerase chain reaction (AC-PCR).Methods: HSV-1 or HSV-2 was captured by anti-HSV glycoprotein monoclonal antibodies coating to polypropylene micro-centrifugal tubes. One type-common upstream primer and two type-specific downstream primers were prepared to amplify DNA from HSV-1 or HSV-2.Results:By direct gel analysis, the products of standard HSV-1 and HSV-2 strains had the expected sizes of 477 bp and 399 bp, respectively. The sensitivity of AC-PCR was very high. 10 PFU HSV-1 or 1 PFU HSV-2 could be detected and typed. AC-PCR was used to detect HSV DNA in 365 swabs from the female genital lesions. One hundred one cases were positive, with 23 cases of HSV-1 (22.8%) and 78 cases of HSV-2 (77.2%). One hundred and twelve of the 365 swab specimens were tested for HSV by both AC-PCR and viral culture. The positivity rate of AC-PCR was higher than that of viral culture (26.8% vs 20.5%, P<0.05). Conclusion : AC-PCR is a specific, sensitive and rapid method for the detection and typing of HSV DNA in genital specimens from female patients.