Taqman MGB荧光PCR检测生殖支原体  

Development of a Taqman MGB real time fluorescence-based PCR assay for the detection of Mycoplasma gerdtalium

作　　者：向志[1] 尹跃平[1] 施美琴[1] 王红春[1] 韩燕[1] 王宝玺[1] 陈祥生[1] 

机构地区：[1]中国医学科学院北京协和医学院皮肤病研究所,南京210042

出　　处：《中华皮肤科杂志》2012年第5期349-351,共3页Chinese Journal of Dermatology

基　　金：国家十一五“重大新药创制”科技重大项目专项（2009ZX09303-005）;“艾滋病和病毒性肝炎等重大传染病防治”科技重大专项（2008ZX10001-005）

摘　　要：目的参照文献建立一种快速、灵敏、准确的生殖支原体（Mg）的检测方法，并应用该方法调查生殖支原体在广西贺州暗娼中的流行情况。方法参考文献中以生殖支原体Pa基因为靶基因设计的引物和Taqman MGB探针进行实时定量PCR，以生殖支原体标准菌株G37制备标准品，对广西贺州暗娼人群的宫颈拭子标本进行生殖支原体的检测。结果建立的Taqman MGB实时定量PCR方法检测的线性范围好（1×10—10^6拷贝／μl，R^2=0．993，重复性好，（批内变异系数=0．7％，批间变异系数：1．09％），敏感性高，检测限（LOD）为10拷贝／μl，定量限（LOQ）为50拷贝／μl。采用该方法检测404份拭子标本，生殖支原体的检出率为12．1％。结论针对生殖支原体Pa基因的TaqmanMGB实时PCR可以快速、灵敏地对生殖支原体进行定性及定量的检测。Objective To establish a rapid, sensitive and accurate method to detect Mycoplasma genitalium, and to evaluate the prevalence of M. genitalium among unlicensed prostitutes from Hezhou city in Guangxi Zhuang Autonomous Region. Methods A pair of primers and Taqman MGB probe were designed and synthesized for the Pa gene of M. genitalium. Standard samples were prepared with the M. genitalium type strain G37. The established Taqman MGB real time fluorescence-based PCR assay was used to detect M. genitalium in the standard samples and cervical swab specimens collected from unlicensed prostitutes in Hezhou city of Guangxi Zhuang Autonomous Region. Results The established Taqman MGB real time PCR exhibited a wide linear range （1× 10 copies/μl to 1 × 10^6 copies/μl, R^2 = 0.993）, good repeatability （intra-assay variation： 0.7%, inter-assay variation： 1.09%） and high sensitivity with the limit of detection being 10 copies/μl and limit of quantification being 50 copies/μl. As the assay showed, 12.1% of the 404 cervical swab samples were positive for M. genitalium. Conculsion The Taqman MGB real time fluorescence-based PCR is a rapid and sensitive method for the quantitative and qualitative detection of M. genitalium.

关 键 词：TaqmanMGB实时PCR 生殖支原体 检测限 荧光PCR 实时定量PCR方法 宫颈拭子标本 参考文献 暗娼人群 

分 类 号：R759[医药卫生—皮肤病学与性病学]