聚合酶链反应扩增指纹多态性快速分型技术在部队结核病分子流行病学研究中的应用

李旭东; 黄悦勤; 李立明; 王玉凤

Abstract

李旭东,黄悦勤,李立明,王玉凤.聚合酶链反应扩增指纹多态性快速分型技术在部队结核病分子流行病学研究中的应用[J].Chinese journal of Epidemiology,2003,24(5):337-380

聚合酶链反应扩增指纹多态性快速分型技术在部队结核病分子流行病学研究中的应用

Study on molecular epidemiology of Mycobacterium tuberculosis in Chinese army with PCR amplified fingerprinting methods

Received:September 10, 2002

DOI：

KeyWord: 结核杆菌 DNA指纹法流行病学,分子

English Key Word: Mycobacterium tuberculosis DNA fingerprints Epidemiology,molecular

FundProject:全军青年基金资助项目 (98Q020 )

Author Name	Affiliation
LI Xu-dong	Department of Preventive Medicine, School of Public Health, Peking University, Beijing 100083, China
HUANG Yue-qin	Department of Preventive Medicine, School of Public Health, Peking University, Beijing 100083, China
LI Li-ming	Department of Preventive Medicine, School of Public Health, Peking University, Beijing 100083, China
WANG Yu-feng	Department of Preventive Medicine, School of Public Health, Peking University, Beijing 100083, China

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Abstract:

目的探讨南方部队结核病的分子流行病学规律。方法设计一对特异性IS6110外向性引物,应用聚合酶链反应(PCR)建立检测结核分支杆菌DNA指纹多态性的方法,分析结核分支杆菌DNA多态性与流行病学的关系。结果共检测分析了154株结核分支杆菌DNA的指纹多态性。根据这些菌株的指纹多态性特征共分为8个类型,以Ⅰ型(36.4%)、Ⅱ型(31.8%)和Ⅲ型(21.4%)为主,其余各型均少于4%。以20～29岁和30～39岁组在这三型中所占比例最大,分别为31.8%和27.9%。驻城镇部队与驻乡村部队以及结核病患者有无卡介苗接种史,在这三型的分布差异无显著统计学意义(P＞0.05)。但初治与复治患者分离菌株的PCR扩增指纹类型的分布差异有显著统计学意义(P＜0.05)。所检测菌株是否具有耐药性,在这三型中的分布差异也有显著统计学意义(P＜0.05)。菌株耐药主要为单耐异烟肼和利福平,耐药菌株在Ⅰ、Ⅱ、Ⅲ型中所占比例分别是44.4%、29.6%和14.8%。结论PCR扩增指纹多态性分型技术是一种简便、快速、敏感、特异和重复性好的分型方法,可用于结核病的分子流行病学研究。南方部队结核分支杆菌的传播以Ⅰ、Ⅱ、Ⅲ型为主,应加强此三型菌株流行的监控。

English Abstract:

Objective　Typing of Mycobacterium tuberculosis strains and epidemiological studies in the army of southern China to provide scientific basis for prevention of pulmonary tuberculosis.MethodsA rapid fingerprinting of M.tuberculosis strains method by pdymerase chain reaction (PCR)with outward-directed primers that designed to the ends of the insertion sequence IS6110 was developed,and to analyzethe relationship between the polymorphism of DNA fingerprinting and epidemiology of M.tuberculosis.Results One hundred and fifty-four M.tuberculosis detected were classified into eight types according to their characters of PCR amplified fingerprints.The main types were typeⅠ(36.4%),type Ⅱ (31.8%),and type Ⅲ (21.4%),while other types w ere less than 4 percentage.In those main type groups,patients aged 20 to 29 and 30 to 39 took up 31.8% and 27.9% respectively.For those main types,the distribution of those types in the first treated patients showed significant difference compared with that in the retreated patients,and the rate of drug-resistance was also statistically different.However,the distribution was not statistically significant to history of BCG vaccination and patients living in urban or rural area.The main drug-resistant strains were only Isoniazid-resistant or Rifampin-resistant strains,while the drug-resistant strains were 44.4%,29.6% and 14.8% respectively in type Ⅰ,type Ⅱ and type Ⅲ.Conclusion　PCR fingerprinting was a rapid, precise,sensitive, specific method to type M.tuberculosis,and could be used to study the epidemiology of tuberculosis;The prevalence of tuberculosis was primarily due to the transmission of type Ⅰ,type Ⅱ and type Ⅲ in the army being studied from Southern China,to suggest that surveillance needs to be strengthened.

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