| 环介导等温扩增技术快速检测结核分枝杆菌核酸 |
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| 引用本文： | 沈会平,张耀祺,杨坚,石磊,陈涛,李国周,赵红波,莫自耀. 环介导等温扩增技术快速检测结核分枝杆菌核酸[J]. 临床检验杂志, 2011, 29(5): 378-380 |
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| 作者姓名： | 沈会平 张耀祺 杨坚 石磊 陈涛 李国周 赵红波 莫自耀 |
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| 作者单位： | 华南理工大学轻工与食品学院，广州 510640;华南理工大学轻工与食品学院，广州 510640;迪澳生物科技有限公司，广州 510663;华南理工大学轻工与食品学院，广州 510640;广东省结核病防治研究所，广州 510630;东莞市慢性病防治院，广东东莞 523008;广州医学院呼吸疾病国家重点实验室，广州 510230;广州医学院呼吸疾病国家重点实验室，广州 510230 |
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| 基金项目： | 呼吸疾病国家重点实验室开放课题（2007DA780154F0904）；广州经济技术开发区科技项目（20090723000210000015）。 |
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| 摘 要： | 摘要:目的：建立一种快速准确的检测结核分枝杆菌（MTB）核酸的环介导等温扩增（LAMP）方法。 方法：以重复插入序列IS6110为目的基因，设计LAMP引物，特异检测MTB核酸。用本法与痰涂片抗酸染色镜检法、实时荧光PCR法对100例可疑患者痰标本进行对比检查。 结果：LAMP法特异性强，仅扩增MTB复合群核酸；灵敏度高，检测限达100 fg；而实时荧光PCR检测限为1 pg。对100例疑似结核病患者痰液标本检测，涂片抗酸染色法、LAMP法、实时荧光PCR法的阳性率分别为28%、39%和38%。 结论：本研究建立的LAMP方法检测MTB核酸特异性强、灵敏度高、时间短且操作简便，有望成为临床快速检测MTB的新方法。
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| 关 键 词： | 结核分枝杆菌；环介导等温扩增法；IS6110基因；实时浊度仪 |
| 修稿时间： | 2011-05-31 |
Rapid detection for Mycobacterium tuberculosis by loop-mediated isothermal amplification |
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| SHEN Hui-ping,ZHANG Yao-qi,YANG Jian,SHI Lei,CHEN Tao,LI Guo-zhou,ZHAO Hong-bo,MO Zi-yao(.College of Light Industry , Food Science,South China University of Technology,Guangzhou ,Guangdong,.Diao Bio-Technology Co.Ltd,Guangzhou ,.Guangdong Research Institute for Mycobacterium tuberculosis Control,Guangzhou . Rapid detection for Mycobacterium tuberculosis by loop-mediated isothermal amplification[J]. Chinese Journal of Clinical Laboratory Science, 2011, 29(5): 378-380 |
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| Authors: | SHEN Hui-ping ZHANG Yao-qi YANG Jian SHI Lei CHEN Tao LI Guo-zhou ZHAO Hong-bo MO Zi-yao(.College of Light Industry Food Science South China University of Technology Guangzhou Guangdong .Diao Bio-Technology Co.Ltd Guangzhou .Guangdong Research Institute for Mycobacterium tuberculosis Control Guangzhou |
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| Affiliation: | SHEN Hui-ping1,ZHANG Yao-qi1,YANG Jian2,SHI Lei1,CHEN Tao3,LI Guo-zhou4,ZHAO Hong-bo5,MO Zi-yao5(1.College of Light Industry and Food Science,South China University of Technology,Guangzhou 510640,Guangdong,2.Diao Bio-Technology Co.Ltd,Guangzhou 510663,3.Guangdong Research Institute for Mycobacterium tuberculosis Control,Guangzhou 510630,4.Dongguan Hospital for Chronic Disease,Dongguan 523008,5.The State Key Laboratory of Respiratory Diseases,Guangzhou Medical University,Guangzh... |
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| Abstract: | Objective To develop a rapid loop-mediated isothermal amplification(LAMP) method for detecting the nuclear acid of Mycobacterium tuberculosis(MTB).Methods The repetitive insertion sequence IS6110 was used as target gene to develop an efficient LAMP method by which MTB was specifically detected.MTB in 100 clinical sputum samples were detected simultaneously by direct smear,LAMP and real-time fluorescent PCR.Results The developed LAMP method showed high specificity for detecting pathogenic strains of MTB comp... |
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| Keywords: | Mycobacterium tuberculosis loop-mediated isothermal amplification IS6110 gene real-time turbidity meter |
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