继发不育男性泌尿生殖道解脲支原体基因分型鉴定及其临床应用研究

Species identification and genotyping of ureaplasma in urogenital tract of male with secondary infertility and its clinical application

目的了解继发不育男性患者泌尿生殖系解脲支原体(UU)的感染状况,探讨UU各基因型与临床病原学之间的关系。方法根据UU多带抗原基因(MBA)与16S-rRNA基因和尿素酶基因结构设计10对引物,应用聚合酶链反应(PCR)技术,对继发不育男性患者572例泌尿生殖系感染的UU进行生物变种和基因型分型鉴定,并与培养结果比较。结果UU培养阳性278例,阳性率48.6% ;PCR基因扩增检测UU-DNA阳性311例,阳性率54.4%。其中生物变种1 (biovar 1)212例,占37.1%;生物变种2(biovar 2)99例,占17.3%;基因分型结果:生物变种1血清变种1 71例,占12. 4%;血清变种3/1498例,占17.2%;血清变种6 43例,占7.5%。生物变种2亚型1(subtype 1)32例,占5.6%;生物变种2亚型2(subtype 2)51例,占8.9%;生物变种2亚型3(subtype 3)16例,占2.8%。结论 UU感染是男性继发不育的主要危险因素,多带抗原基因与16S-rRNA基因和尿素酶基因分型鉴定具有简便、快速、敏感、特异之优点。

Objective： To understand the infection of ureaplasma species in urogenital tract of the males with secondary infertility, and explore the relationship between the species or subtypes and male infertility. Methods： Ten pairs of oligonucleotide primers, targeting 5 ＇ ends of the multiple-banded antigen （MBA） genes, 16S rRNA gene and the urease gene subunits, were designed to identify genotype of the ureaplasma species by PCR-based typing system. These primers could distinguish two biovars and 14 serovars of U. ureaplasma. Results：A total of 278 （48. 6%） positive ureaplasma cultures were obtained from 572 patients attending our center. PCR method revealed U. ureaplasma positive in 311 （54.4%） of 572 patients, with 212 （37.1%） U. parvum （biovar 1） and 99 （17.3%） U. ureaplasma （biovar 2）. Among 212 U. parvum （biovar 1） were 71 （12.4%） serovar 1, 98 （17.1%） serovars3/14 and 43 （7.5%） serovar 6. In 99 U. ureaplasma （biovar 2）, there were 32 （5.6%）, 51 （8.9%） and 16 （2.8%） for subtype 1, subtype 2 and subtype 3, respectively. Conclusion：The infection of ureaplasma in the urogenital tract of males could contribute to the male infertility. The PCR-based genotyping system is relatively rapid, practicable and specific for species identification and subtyping of ureaplasma.