| 结核分枝杆菌荧光定量PCR检测方法建立 |
| |
| 引用本文： | 徐菊玲,徐伯赢,周洪昌,张慧,段劲,薛利军,邵圣文. 结核分枝杆菌荧光定量PCR检测方法建立[J]. 中国公共卫生, 2011, 27(10) |
| |
| 作者姓名： | 徐菊玲 徐伯赢 周洪昌 张慧 段劲 薛利军 邵圣文 |
| |
| 作者单位： | 1. 湖州师范学院医学院创新实验室,浙江湖州,313000
2. 南京军区南京总医院肿瘤内科 |
| |
| 摘 要： | 目的 探讨结核分枝杆菌实时定量检测方法并进行评价.方法针对结核杆菌(Mycobacterium tuberculosis,Mtb)16S rDNA基因设计引物,应用SYBR Green I建立荧光定量PCR( FQ-PCR)反应体系；提取Mtb基因组DNA,PCR扩增16S rDNA片段,构建重组质粒pMD-TB16S;检测11份肺结核患者痰标本；以甲型链球菌、大肠杆菌等基因组DNA作对照,检验方法特异性,对同一份Mtb DNA模板进行批内和批间检测,计算变异系数(CV).结果靶向Mtb 16S rDNA基因引物能够特异扩增Mtb 16S rDNA基因,对照组细菌基因未见扩增,灵敏度为(Mtb基因组DNA) 1.2 pg/μL,即(38.9 +3.54)拷贝/μL的16S rDNA基因,批内和批间Ct值变异系数分别为0.27％和1.26％.结论以16S rDNA为靶基因的FQ-PCR技术,能够对Mtb进行快速、敏感而特异的定量检测.
|
| 关 键 词： | 结核杆菌 16S rDNA 荧光定量PCR |
Establishment of fluorescence quantitative PCR assay in detection of Mycobacterium tuberculosis |
| |
| XU Ju-ling,XU Bai-ying,ZHOU Hong-chang,et al.. Establishment of fluorescence quantitative PCR assay in detection of Mycobacterium tuberculosis[J]. Chinese Journal of Public Health, 2011, 27(10) |
| |
| Authors: | XU Ju-ling XU Bai-ying ZHOU Hong-chang et al. |
| |
| Affiliation: | XU Ju-ling,XU Bai-ying,ZHOU Hong-chang,et al.Laboratory of Innovation,Medical School of Huzhou Teachers College(Huzhou 313000,China) |
| |
| Abstract: | Objective To establish and assess a rapid real-time quantifiable method for the detection of Mycobacterium tuberculosis(Mtb).Methods The specific primers targetting 16S rDNA gene in Mtb were designed and the fluorescence quantitative PCR(FQ-PCR) assay was performed with SYBR Green I.After extraction of Mtb genomic DNA,the 16S rDNA fragment was amplified by conventional PCR and used to construct recombinant pMD-TB16S plasmid.Then,the serial dilutions of pMD-TB16S plasmid were subjected to the quantitation st... |
| |
| Keywords: | Mycobacterium tuberculosis 16S rDNA fluorescence quantitative PCR |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
