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中国农学通报

中国农学通报 ›› 2010, Vol. 26 ›› Issue (10): 1-4.

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

结核杆菌实时荧光定量PCR快速检测方法的建立及应用

孟茹,陈创夫,张辉,乔军,王正荣   

  • 收稿日期:2009-12-23 修回日期:2010-02-22 出版日期:2010-05-20 发布日期:2010-05-20
  • 基金资助:

    国际科技合作重点项目

Construction of Real-time PCR quantitative rapid detection of Mycobacterium tuberculosis

Meng Ru, Chen Chuangfu, Zhang Hui, Qiao Jun, Wang Zhengrong   

  • Received:2009-12-23 Revised:2010-02-22 Online:2010-05-20 Published:2010-05-20

PDF (PC)

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摘要：

针对结核杆菌CFP10基因序列设计并合成了1对引物，以构建的含有该引物扩增序列的重组质粒作为阳性标准品，建立了检测CFP10基因的SYBR GreenⅠreal-time PCR方法。检测结果显示，该方法线性关系好，标准曲线的相关系数达到0.997，对初始模板的检出下限为1 ×101copies/μL，比常规PCR方法高100倍。表明建立的real-time PCR检测方法灵敏度高、特异性强、重复性好，可以用于结核杆菌CFP10的病原检测及定量分析。

关键词: 克伦特罗, 克伦特罗, 心肌, ATP, 抗氧化能力

Abstract:

Mycobacterium tuberculosis CFP10 gene sequence was designed and synthesized primer 1 pairs to build a primer containing the sequence of the recombinant plasmid was amplified as a positive standard, the establishment of a nucleic acid of CFP10 Detection SYBR Green Ｉreal-time PCR method. Test results showed a good linear relationship between the method, standard curve correlation coefficient reached 0.997, the initial template for the detection threshold of 1 × 101copies/μL, than the conventional PCR method of the high 100 times. Show that the established real-time PCR detection method is sensitive, specific, reproducible and can be used to detect the pathogen Mycobacterium tuberculosis CFP10 and quantitative analysis.