Liu Jing-hua, Kremer Kristin, Pourcel Christine, Mulder Arnout, Liu Zhi-guang, Zhao Xiu-qin, Wan Kang-lin
State Key Laboratory for Infectious Diseases Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Zhonghua Liu Xing Bing Xue Za Zhi. 2008 Aug;29(8):801-5.
To develop a standardized IS6110-restriction fragment length polymorphism (RFLP) method, used for evaluating the capacity of genotyping.
IS6110-RFLP of 78 Mycobacterium (M.) tuberculosis strains were studied by bio-molecular techniques including DNA isolation, PCR, restriction endonuclease enzyme analysis, southern blotting, agarose gel electrophoresis, together with data analysis by software Gel-Pro analyzer 3.1 and BioNumerics (Version 5.0).
IS6110-RFLP method was established and standardized successfully, including DNA isolation, PCR, restriction endonuclease enzyme analysis, southern blotting, agarose gel electrophoresis and usage of the analysis software with standard parameters. By this method, 78 M. tuberculosis isolates were classified into 75 genotypes which belonged to 11 different clusters. Of all the isolates, 66.7% (52/78) belonged to a main cluster.
Standard IS6110-RFLP method was established successfully. This method had powerful capacity for genotyping and strain level identification and could be used for the surveillance on pathogens of M. tuberculosis in China.
建立一种标准化的IS6110限制性片段长度多态性（RFLP）方法，用于评估基因分型能力。
采用包括DNA提取、聚合酶链反应（PCR）、限制性内切酶分析、Southern印迹、琼脂糖凝胶电泳等生物分子技术，对78株结核分枝杆菌进行IS6110-RFLP研究，并使用Gel-Pro分析仪3.1和BioNumerics（5.0版）软件进行数据分析。
成功建立并标准化了IS6110-RFLP方法，包括DNA提取、PCR、限制性内切酶分析、Southern印迹、琼脂糖凝胶电泳以及使用具有标准参数的分析软件。通过该方法，78株结核分枝杆菌分离株被分为75种基因型，属于11个不同的簇。所有分离株中，66.7%（52/78）属于一个主要簇。
成功建立了标准化的IS6110-RFLP方法。该方法具有强大的基因分型和菌株水平鉴定能力，可用于我国结核分枝杆菌病原体的监测。
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