中华皮肤科杂志 ›› 2012, Vol. 45 ›› Issue (5): 349-351.

• 研究报道 • 上一篇    下一篇

Taqman MGB荧光PCR检测生殖支原体

向志1,尹跃平2,施美琴3,王红春4,韩燕4,王宝玺2,陈祥生5   

  1. 1. 中国医学科学院皮肤病研究所
    2. 南京 中国医学科学院北京协和医学院皮肤病研究所
    3. 南京医科院皮研所
    4. 中国医学科学院北京协和医学院皮肤病研究所
    5. 医科院皮研所
  • 收稿日期:2011-06-09 修回日期:2011-08-29 出版日期:2012-05-15 发布日期:2012-05-03
  • 通讯作者: 尹跃平 E-mail:yinyp@ncstdlc.org
  • 基金资助:

    国家十一五“重大新药创制”科技重大项目专项“抗感染药物临床前药效评价技术平台”;“艾滋病和病毒性肝炎等重大传染病防治”科技重大专项“防治性病对预防艾滋病的作用研究”

Development of a Taqman MGB real time fluorescence?鄄based PCR assay for the detection of Mycoplasma genitalium

  • Received:2011-06-09 Revised:2011-08-29 Online:2012-05-15 Published:2012-05-03

PDF

46

赞

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 ~ 106拷贝/μl），R2 = 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Taqman MGB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。

关键词: 流行率

Abstract:

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 × 106 copies/μl , R2 = 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.

Key words: prevalence