关键词 单纯疱疹病毒，Ⅱ型；梅毒螺旋体；双引物聚合酶链反应；快速检测

    摘要 目的 建立双引物聚合酶链反应(PCR)用于同时快速检测梅毒螺旋体和单纯疱疹病毒Ⅱ型。方法 基于单纯疱疹病毒II型(HSV-2)的DNA聚合酶基因以及梅毒螺旋体47KD膜蛋白基因序列，自行设计2对特异性寡核苷酸引物，建立了同时检测上述2种病原体的双引物PCR。结果 各特异性引物仅扩增相应的病原体DNA，即单纯疱疹病毒Ⅱ型扩增产物为202bp以及梅毒螺旋体扩增产物为252bp。其他11种生殖道常见微生物及正常人基因组DNA不被扩增。检测了51例临床标本，梅毒螺旋体暗视野显微镜检查阳性检出率为15.7%，双引物PCR阳性检出率为19.6%；二种方法检测结果间比较差异均无统计学意义(P>0.05)。HSV阳性培养率23.6%，双引物PCR阳性率47.1%，双引物PCR阳性检出率显著高于HSV培养且差异有统计学意义(P=0.019)；上述临床标本分别进行了2种病原体的单引物PCR，与双引物PCR对照结果完全一致。结论 我们建立的双引物PCR一次可同时检测2种病原体，且有快速、敏感、特异以及简便的特点。

    SIMULTANEOUS EXAMINATION OF HERPES SIMPLEX VIRUS Ⅱ AND TREPONEMA PALLIDUM BY DOUBLE-PRIMER PCR

    Liu Aiying,Yin Yueping.

    Department of Dermatology and Venereal Disease,Coal General Hospital,Beijing 100028，China

    Key words herpes simplex virus; treponema pallidum; double-primer PCR; fast detection

    Abstract Objective To approach double-primer PCR assay for simultaneous detection of Herpes simplex virus Ⅱ(HSV-2) and Treponema pallidum. Methods According to specific gene sequence of herpes simplex virus Ⅱ DNA Polymerse gene and treponema pallidum 47 kd gene, 2 sets of specific primers were designed and a double-primer PCR assay were developed to detect 2 agents in one test. Results The target DNAs of 2 agents were specifically amplified by theirs specific primers. The DNA fragment length of amplifying HSV-2 was 202 bp, amplifying treponema pallidum was 252 bp; The DNA of other 11 common microbes of genital tract and human gene were not amplified by 2 sets of primers. 51 clinical samples were detected by dark-field microscopy and a double-primer PCR, the positive rates were 15.7% and 19.6%, respectively. And there was no statistically significant different between dark-field microscopy and a double-primer PCR (P>0.05). Detected by HSV culture and a double-primer PCR, the positive rates were 23.6% and 47.1% respectively. The sensitivity of HSV detected by PCR and a double-primer PCR was significantly higher than that of culture, and there was significant different between HSV culture and PCR and a double-primer PCR (P=0.019). All specimens were tested for HSV and Treponema pallidum by single-primer PCR respectively, the results were consistent to double-primer PCR. Conclusion Primary study indicates that the double-primer PCR assay can simultaneously, specifically, rapidly, sensitively detect 2 agents. ......