摘要
目的应用PCR技术对解脲脲支原体(Uu)多带抗原进行分型鉴定,探讨Uu各种基因型与临床病原学之间的关系。方法根据解脲脲支原体多带抗原基因(MBA)与16S rRNA基因和尿素酶基因结构设计10对引物,采用PCR基因扩增技术,对384例非淋菌性尿道炎和其他泌尿生殖道感染患者的临床标本,进行解脲脲支原体生物变种和基因型分型鉴定,并与Uu培养法作比较。结果384例性病门诊患者临床标本中,检测Uu培养阳性218例,阳性率56.8%;PCR基因扩增检测Uu-DNA阳性208例,总阳性率为54.2%:其中生物变种1(biovar 1)143例,占37.2%,生物变种2(biovar 2)65例,占16.9%;基因分型结果:生物变种1血清变种1(serovar 1)50例,占13.0%,血清变种3/14(serovars 3/14)64例,占16.7%,血清变种6(serovar 6)29例,占7.6%;生物变种2亚型1(subtype 1)25例,占6.5%,亚型2(subtype 2)30例,占7.8%,亚型3(subtype 3)10例,占2.6%。结论Uu是性病的重要病原体,MBA多带抗原PCR基因分型鉴定具有简便、快速、敏感、特异之优点。
OBJECTIVE Ureaplasma urealyticum is a recognized cause of nongonococcal urethritis. It has also been implicated in other genitourinary syndromes. There are two biovars and 14 serovars of U. urealyticum. The MBA （multiple-banded antigen） is the predominant antigen recognized during U. urealyticum infections and is probably an important virulence determinant. The aim of this study is to identify the MBA genes of U. parvum and U.urealyticum by PCR-based typing system. METHODS Urethral and cervical swabs of 384 patients with nongonococcal urethritis and other genitourinary infection in sexually transmitted disease （STD） clinic were collected from various regional hospitals in Guangdong Province to detect genotyping methods. The cultures were performed as soon as possible after receipt of specimens in the laboratory, while biovar and subtyping of MBA gene were performed by PCR. In this study, we designed 10 pairs of oligonucleotide primers, targeting the 5＇ ends of the MBA genes, to identify and genotype these Ureaplasrna species. The 10 primer pairs could distinguish the two species, and subtypes within each species. RESULTS A total of 218 （56.8%） positive Ureaplasrna culture were obtained from 384 patients attending a STD clinic. These methods were used to identify and genotype U.urealyticurn in 208 （54.2 %） of 384 patients with genitourinary infection. Among them U. parvurn （biovar 1） was detected in 37.2% and U. urealyticurn （biovar 2） in 16.9%. A selection of 10 primer pairs used to identify and genotype, the results showed that serovar 1 was in 13.0%, serovars 3/14 in 16.7%, serovar 6 in 7.6% ; subtype 1 of serovar 2 was in 6.5%, subtype 2 in 7.8% and subtype 3 in 2.6%, respectively. CONCLUSIONS The PCR-based genotyping system will facilitate future studies of the relationship between individual Ureaplasrna species or subtypes and human disease. The methods described are relatively rapid, practicable, and specific for the detection, species identification and subtyping of Ureaplasma spe
出处
《中华医院感染学杂志》
CAS
CSCD
北大核心
2005年第8期856-858,891,共4页
Chinese Journal of Nosocomiology
基金
广州市教委99年重点专项资助(0705-B047)
关键词
解脲脲支原体
多带抗原
基因型
聚合酶链反应
Ureaplasma urealyticum Multiple-banded antigen （MBA） Genotypes Polymerase chain reaction
