巢式聚合酶链反应检测男性尿道炎患者尿液中阴道毛滴虫

Detection of Trichomonas vaginalis in male patients with urethritis by nested PCR

目的 建立两种巢式聚合酶链反应（PCR）方法检测男性尿道炎患者尿液中阴道毛滴虫感染状况,评价两种巢式PCR在临床诊断中的应用价值.方法 2011年4月至2013年12月来我院性病门诊就诊的1 088例男性尿道炎患者为研究对象,收集尿道拭子标本做分泌物涂片镜检、阴道毛滴虫湿片检测以及淋球菌培养,同时收集尿液标本提取DNA,针对阴道毛滴虫重复基因组和β微管蛋白基因,采用两种巢式PCR法检测尿液中阴道毛滴虫.结果 湿片法检测阴道毛滴虫的阳性率为0,而两种巢式PCR法均检测出29例阳性标本,阳性率为2.67％,且两种巢式PCR法检测出的阳性标本一致.结论 与湿片法相比,巢式PCR法检测男性尿液标本阴道毛滴虫具有较高的灵敏度和特异性.

Objective To establish two nested PCR assays for detection of Trichomonas vaginalis in urine samples from male patients with urethritis,and to evaluate their diagnostic value.Methods One thousand and eighty-eight male patients with urethritis were enrolled from sexually transmitted disease （STD） clinic in the Hospital of Dermatology,Chinese Academy of Medical Sciences and Peking Union Medical College between April 2011 and December 2013.Urethral swabs were collected followed by smear testing,wet mount microscopic examination of Trichomonas vaginalis,and cultivation of Neisseria gonorrhoeae.Urine specimens were also obtained from these patients followed by DNA extraction.Two nested PCR assays were developed and performed to amplify the repeat genomic sequence and β-tubulin gene of Trichomonas vaginalis.Results Trichomonas vaginalis was detected in none of these swab specimens by wet mount microscopy,but in 29 （2.67%） of the urine specimens by either of the two nested PCR assays.Moreover,the positive specimens detected by the two nested PCR assays were completely consistent.Conclusion Compared with wet mount microscopy,nested PCR has higher sensitivity and specificity in detection of Trichomonas vaginalis in urine samples from male patients.