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中国结核分枝杆菌间隔区寡核苷酸分型方法标准化操作程序的探讨
  • 期刊名称:中华流行病学杂志
  • 时间:0
  • 页码:384-387
  • 语言:中文
  • 分类:R378.911[医药卫生—病原生物学;医药卫生—基础医学] R392.1[医药卫生—免疫学;医药卫生—基础医学]
  • 作者机构:[1]中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室,北京102206
  • 相关基金:基金项目:国家自然科学基金(30771853)
  • 相关项目:中国结核分枝杆菌基因多态性及其株水平鉴定技术研究
作者: 万康林|
中文摘要:

目的探讨中国结核分枝杆菌间隔区寡核苷酸分型(Spoligotyping)的标准化方法,初步评价其应用价值。方法采用核酸提取、聚合酶链反应(PCR)、反向线性点杂交等技术,结合BioNumerics(Version5.0)软件,对224株结核分枝杆菌临床分离株进行分型研究。结果使用Spoligotyping标准化方法对224株中国结核分枝杆菌临床分离菌株进行基因分型,将其分为北京家族菌株和非北京家族菌株两大簇。其中北京家族菌株129株,为主要的流行菌株。结论初步确定中国Spoligotyping标准化技术方案。该方法快速、简便、重复性好,能同时对结核分枝杆菌进行检测和分型,有利于结核病传染源的追溯和流行趋势的研究,尤其是在鉴定北京家族菌株上具有独特作用。

英文摘要:

Objective To study and preliminarily evaluate the standard spacer oligonucleotide typing (Spoligotyping) method and the application on Mycobacterium tuberculosis (M.tuberculosis). Methods Spoligotyping on 224 M.tuberculosis strains was studied by the molecular biological techniques, including DNA isolation, PCR, reverse line blot hybridization, together with data analysis software BioNumerics (Version 5.0). Results Standardization on both Spoligotyping method and parameters in result analysis and the usage of the analysis software were studied. Through this method, 224 M.tuberculosis clinical strains were classified into 2 clusters including 129 Beijing family strains and 95 non-Beijing family strains. The predominant strains belonged to Beijing family. Conclusion Standard Spoligotyping method was preliminary determined in China, showing that it was a simple, rapid, and robust method for simultaneous detection and typing of M.tuberculosis. This method can be used for tracing the source of infection and understanding the epidemic trend of M.tuberculosis. Spoligotyping can also be served as a method for simultaneous detection and typing of M.tuberculosis, and to identify Beijing family strains.

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