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解脲脲支原体多带抗原基因分型鉴定及其临床应用研究

Multiple-Banded Antigen for Ureaplasma by Polymerase Chain Reac tion:Their Species Identification and Genotyping and Clinical Application

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【作者】 朱庆义胡朝晖刘峰林刘懿微郑建树汪凡军

【Author】 ZHU Qing-yi,HU Zhao-hui,LIU Feng-lin,LIU Yi-wei,Z HENG Jian-shu,WANG Fan-jun(Guangzhou Kingmed Center for Clinical Laboratory,Guangzh ou 510330,China)

【机构】 广州金域医学检验中心佛山市顺德区均安医院广州金域医学检验中心 广东广州510330广东广州510330广东佛山528500广东广州510330

【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。

【Abstract】 OBJECTIVE Ureaplasma u re alyticum is a recognized cause of nongonococcal urethritis.It has also been implicated in other genitourinary syndromes.There are two biovars and 14 s erovars of U.urealyticum.The MBA(multiple-banded antigen) is the predominant antigen recognized during U.urealyticum infectio ns 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 38 4 patients with nongonococcal urethritis and other genitourinary infection in se xually transmitted disease(STD) clinic were collected from various regional hosp itals in Guangdong Province to detect genotyping methods.The cultures were perf ormed as soon as possible after receipt of specimens in the laboratory,while bi ovar 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,t o identify and genotype these Ureaplasma species.The 10 primer pairs could distinguish the two species,and subtypes within each species. RESULTS A total of 218(56.8%) positive Ureaplasma culture were obtained from 384 patients attending a S TD clinic.These methods were used to identify and genotype U.urealytic um in 208(54.2%) of 384 patients with genitourinary infection.Among t hem U.parvum(biovar 1) was detected in 37.2% and U.ure alyticum(biovar 2) in 16.9%.A selection of 10 primer pairs used to id entify 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 sy stem will facilitate future studies of the relationship between individual Ureaplasma species or subtypes and human disease.The methods describ ed are relatively rapid,practicable,and specific for the detection,species id entification and subtyping of Ureaplasma species.

【基金】 广州市教委99年重点专项资助(0705-B047)
  • 【文献出处】 中华医院感染学杂志 ,Chinese Journal of Nosoconmiology , 编辑部邮箱 ,2005年08期
  • 【分类号】R375;
  • 【被引频次】26
  • 【下载频次】143
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