男性泌尿生殖道感染患者奈瑟菌属菌种的基因检测  

Gene detection of Neisseria spp.isolated from male patients with genitourinary tract infections

作　　者：王和[1] 罗振华[1] 易旭[1] 佘晓玲[1] 王丹霓[1] 王艳[2] 叶长芸[2] 

机构地区：[1]贵阳医学院微生物学教研室,贵州贵阳550004 [2]中国疾病预防控制中心传染病预防控制所,北京102206

出　　处：《中华男科学杂志》2012年第1期39-43,共5页National Journal of Andrology

基　　金：贵州省科技基金(SY[2001]3045);传染病预防控制国家重点实验室自主研究开放课题基金(2011SKLID302);贵州省卫生厅科技基金(gzwkj2009-1-028)~~

摘　　要：目的:检测从男性泌尿生殖道感染患者分离的奈瑟菌属菌种的基因并探讨非淋病奈瑟菌的致病性以及奈瑟菌属菌种感染的实验室诊断方法。方法:用聚合酶链反应及核苷酸序列测定方法,检测男性泌尿生殖道感染患者泌尿生殖道分离的奈瑟菌属菌种的淋病奈瑟菌核酸扩增荧光检测试剂的反应及其染色体上16SrRNA基因、淋病奈瑟菌DNA甲基转移酶基因orf1、隐蔽性质粒基因cppB及外膜蛋白基因nspA。结果:男性泌尿生殖道感染患者泌尿生殖道分离的14株奈瑟菌属菌种经16SrRNA基因检测鉴定为淋病奈瑟菌2株、粘液奈瑟菌3株、灰色奈瑟菌3株、干燥奈瑟菌2株、微黄奈瑟菌2株、嗜乳糖奈瑟菌1株、多糖奈瑟菌1株,其中淋病奈瑟菌核酸扩增荧光检测试剂阳性反应的9株、cppB阳性反应1株、orf1阳性反应5株、nspA阳性反应3株。16SrRNA基因检测与常规细菌学方法鉴定结果比较,两者的符合率85.7%。结论:引起男性泌尿生殖道感染的非淋病奈瑟菌缺乏cppB,大多数菌株缺乏淋病奈瑟菌毒力相关的orf1与nspA并且可在淋病奈瑟菌核酸扩增荧光检测试剂检测中显示阳性反应,从而造成淋病奈瑟菌鉴定与淋病诊断的误诊以及其他奈瑟菌感染的漏诊。在疑似奈瑟菌属菌种感染的实验室检查中联合使用常规细菌学方法与基因检测方法,可有助于提高奈瑟菌属菌种鉴定及其感染诊断的准确率。Objective： To detect the genes of Neisseria spp.isolated from patients with male genitourinary tract infections,and to study the pathogenicity of non-gonococcal strains of Neisseria and the laboratory diagnosis for the infections caused by Neisseria spp.Methods： Using polymerase chain reaction and nucleotide sequencing,we amplified and sequenced 4 genes of Neisseria spp.isolated from patients with male genitourinary tract infections,including 16S rRNA,orf1,cppB and nspA.Results： Fourteen Neisseria strains were identified through analysis of the 16S rRNA gene,including 3 N.mucosa strains,3 N.cinerea strains,2 N.gonorrhoea strains,2 N.sicca strains,2 N.subflava strains,1 N.lactamica strain,and 1 N.polysaccharea strain.Among them,9 showed positive results in gonococcal fluorescence-labeled multiplex-PCR detection,1 in cppB gene reaction,5 in orf1 gene reaction,and 3 in nspA gene reaction.The consistency rate was 85.7% between the above results from our gene detection and those from the routine bacteriological methods.Conclusion： The cppB gene is absent in the non-gonococcal strains of Neisseria spp.that can cause male genitourinary tract infection.Most of the strains not only lack virulence-associated orf1 and nspA genes,but also show positive results in gonococcal fluorescence-labeled multiplex-PCR detection,which is one of the important reasons for the misdiagnosis and missed diagnosis of gonorrhea infection.The combination of routine bacteriological methods and gene detection in laboratory examinations may help improve the accuracy rates of Neisseria species identification and clinical diagnosis of the infections caused by Neisseria spp.

关 键 词：奈瑟菌属 淋病 外膜蛋白质 基因 诊断 

分 类 号：R440[医药卫生—诊断学]