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| 定量PCR快速检测炭疽芽孢杆菌的实验研究 | |
| 引用本文： | 曲识,史清海,何宁,陈永胜,周蕾,郭兆彪,周冬生,翟俊辉,杨瑞馥.定量PCR快速检测炭疽芽孢杆菌的实验研究[J].军事医学科学院院刊,2010,34(3):275-279. |
| 作者姓名： | 曲识 史清海 何宁 陈永胜 周蕾 郭兆彪 周冬生 翟俊辉 杨瑞馥 |
| 作者单位： | 1. 军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京,100071 2. 公安部消防局,北京,100054;北京科技大学土木与环境工程学院,北京,100083 3. 上海市消防总队特勤支队,上海,200127 |
| 摘 要： | 目的建立一种简便、快速、定量检测炭疽芽孢杆菌的PCR方法。方法采用TaqMan荧光探针技术,针对炭疽芽孢杆菌质粒pXO1、pXO2上的基因设计引物;用炭疽芽孢杆菌（Sterne株）污染环境土壤来模拟实际样本,比较了从土壤中提取DNA的不同方法对检测效果的影响,并与同类进口试剂进行了平行比对。结果以含有检测目的片段的线性质粒为模板,反应体系的灵敏度可达到101拷贝/μl;用Sterne株芽孢悬液污染土壤,检测灵敏度可达2.5×103cfu/g土壤。我们研制的试剂与进口试剂在敏感性和准确性方面一致,且操作简便。结论本研究建立的炭疽芽孢杆菌实时定量PCR检测方法可特异、灵敏地检测土壤标本中可疑目标菌。 |
| 关 键 词： | 炭疽芽孢杆菌 实时荧光定量PCR TaqMan探针 |
Real time quantitative PCR and its reagent product for rapid detection of Bacillus anthracis |
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| QU Shi,SHI Qing-hai,HE Ning,CHEN Yong-sheng,ZHOU Lei,GUO Zhao-biao,ZHOU Dong-sheng,ZHAI Jun-hui,YANG Rui-fu.Real time quantitative PCR and its reagent product for rapid detection of Bacillus anthracis[J].Bulletin of the Academy of Military Medical Sciences,2010,34(3):275-279. | |
| Authors: | QU Shi SHI Qing-hai HE Ning CHEN Yong-sheng ZHOU Lei GUO Zhao-biao ZHOU Dong-sheng ZHAI Jun-hui YANG Rui-fu |
| Institution: | 1.Laboratory of Analytical Microbiology,State Key Laboratory of Pathogen and Biosecurity,Institute of Microbiology and Epidemiology,Academy of Military Medical Sciences,Beijing 100071,China;2.Fire Department of Ministry of Public Security,Beijing 100054,China;3.Civil and Environmental Engineering School,Beijing University of Science and Technology,Beijing 100083,China;4.The Special Branch of Shanghai Fire Bureau,Shanghai 200127,China) |
| Abstract: | Objective To establish a simple and rapid quantitative real time PCR method for detecting Bacillus anthracis with LightCycler.Methods An assay for detection of B.anthracis was developed using TaqMan technology with the primers and probes targeting sequences from genes of the plasmids pXO1 and pXO2,respectively.The assays were performed using soil samples artificially contaminated with B.anthracis strain Sterne.Our reagent was compared with the imported one in terms of sensitivity and accuracy.Results The sensitivity of the real time PCR reached 101 copies/μl using linear recombinant plasmid as template and 2.5×103 spores per gram soil using contaminated soil samples.Our reagent was similar to the imported one in terms of sensitivity and accuracy,but took less time to treat samples.Conclusion A sensitive and specific real time PCR method based on plasmid genes is developed for rapid detection and identification of B.anthracis in clinical samples,such as soil. |
| Keywords: | Bacillus anthracis real-time quantitative PCR TaqMan probe |
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