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| 结核杆菌多位点环介导等温扩增检测方法的建立 | |
| 引用本文： | 王正荣,陈创夫,杨明锋,孟茹,曹旭东,张辉,乔军.结核杆菌多位点环介导等温扩增检测方法的建立[J].中国预防兽医学报,2010,32(4). |
| 作者姓名： | 王正荣 陈创夫 杨明锋 孟茹 曹旭东 张辉 乔军 |
| 作者单位： | 1. 石河子大学,动物科技学院,新疆,石河子,832003 2. 石河子大学,动物科技学院,新疆,石河子,832003;新疆地方与民族高发病教育部重点实验室,新疆,石河子,832003 3. 新疆地方与民族高发病教育部重点实验室,新疆,石河子,832003 |
| 基金项目： | 国家十一五传染病重大专项，国际科技合作项目 |
| 摘 要： | 为建立一种快速、高敏感、高特异、鉴别结核分枝杆菌和牛分枝杆菌的方法,本研究根据大多数致病性结核分枝杆菌共有序列esat-6,结核分枝杆菌和牛分枝杆菌gyrB共有特异性序列,以及结核分枝杆菌种特异序列mtp40设计6对特异性引物对痰液中的结核分枝杆菌进行检测,并与鉴别培养基分离培养结果以及常规PCR鉴定结果进行比较。实验结果表明,建立的LAMP检测方法具有很高的特异性。可区分致病性结核分枝杆菌和非致病结核分枝杆菌,也可鉴别结核分枝杆菌和牛分枝杆菌。LAMP检测技术的灵敏度比经典PCR技术高100倍左右,可检测到7拷贝/反应。另外,用3种方法同时检测样品发现,LAMP与细菌培养的符合率为90.91%,LAMP与常规PCR检测结果的符合率为100%。LAMP检测方法可以在流行病学调查及现场快速诊断方面广泛应用,并为临床治疗提供依据。 |
| 关 键 词： | 环介导等温扩增 结核分枝杆菌 牛分枝杆菌 鉴别诊断 |
Development of multi-site loop-mediated isothermal amplification (LAMP) for rapid detection of Mycobacterium tuberculosis |
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| WANG Zheng-rong,CHEN Chuang-fu,YANG Ming-feng,CAO Xv-dong,ZHANG Hui,QIAO Jun.Development of multi-site loop-mediated isothermal amplification (LAMP) for rapid detection of Mycobacterium tuberculosis[J].Chinese Journal of Preventive Veterinary Medicine,2010,32(4). | |
| Authors: | WANG Zheng-rong CHEN Chuang-fu YANG Ming-feng CAO Xv-dong ZHANG Hui QIAO Jun |
| Institution: | WANG Zheng-rong1,CHEN Chuang-fu1,2,YANG Ming-feng1,CAO Xv-dong2,ZHANG Hui1,QIAO Jun1(1. College of Animal Science , Technology,Shihezi University,Shihezi 832003,China,2. Ministry of Education Key Laboratory of Xinjiang Endemic , Ethnic Disease,China) |
| Abstract: | To develop an efficient,sensitive and specific method for detection of Mycobacterium tuberculosis and Mycobacterium bovis,specifc primers were designed for a multi-site loop-mediated isothermal amplification(LAMP) method by targeting the esat-6 gene,gyrB gene and mtp40 gene. The results of LAMP detection were compared with that of bacterium isolation and PCR detection. The LAMP is highly efficient and specific in detection of M. tuberculosis and M. bovis and could detect 7 copies/reaction,which was 100 time... |
| Keywords: | Loop-mediated isothermal amplification Mycobacterium tuberculosis Mycobacterium bovis identical dignosis |
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