结核分枝杆菌随机扩增多态性DNA指纹分型技术及应用研究

Fingerprinting by random amplified polymorphic DNA of M tuberculosis and its application

目的 探讨重庆地区结核病分子流行病学的发展趋势。方法 建立随机扩增多态性DNA聚丙酰胺凝胶电泳 (RAPD PAGE)指纹分型法 ;采用 1对随机引物对重庆地区 2 87例肺结核患者41 6株结核分枝杆菌进行DNA指纹分型。结果 41 6株分离菌株其DNA指纹可分为 7种类型 ,以Ⅰ型 (35 .5 % )、Ⅱ型 (2 9 3 % )和Ⅲ型 (2 2 .6 % )为主 ,其中 2 0～ 39岁和 40～ 49岁年龄组的初治患者在 3型中分别占 58.0 %和 46 .2 %。在Ⅰ、Ⅱ、Ⅲ型中初治患者分别占 52 .0 %、46 .4%和 58.5 % ,至少耐利福平和异烟肼 (MDR)的初治患者又分别占的 1 1 .3 %、35 .9%和 2 4 .3 % ,至少耐利福平和其他药物的多重耐药株在 3型中占 2 1 .7% (初治 )。结论 RAPD PAGE指纹分型法 ,是一种简便、特异、敏感、重复性好的菌株分型方法 ,可用于结核病分子流行学的研究 ;以Ⅰ、Ⅱ、Ⅲ型为主的结核分枝杆菌正在重庆及周边地区的人群中传播 ;

Objective Epidemiological studies on Mycobacterium tuberculosis in Chongqing. Methods To establish a fingerprinting method by random amplified polymorphic DNA and polyacrylamide gel electrophoresis (RAPD PAGE). The fingerprints of genomic DNA of M tuberculosis strains, which were isolated from 416 sputum specimens among the 287 lung tuberculosis patients, were detected by RAPD PAGE fingerprinting with primer 1 (5′ CCGGGGCGGTTC) and primer 2 (5′ CCGCCGACCGAC). Results Seven types of DNA fingerprints were found and the main types were type Ⅰ (35.5%), type Ⅱ (29.3%) and type Ⅲ (22.6 %). In those main type groups, the first treated patients aged from 20 to 39 and from 40 to 49 were 58.0% and 46.2% respectively. For type Ⅰ, type Ⅱ and type Ⅲ, the first treated patients were 52.0%, 46.4% and 58.5%, the first treated with the anti tuberculosis chemicals, the multi drug resistant (MDR) strains were 11.3%, 35.9% and 24.3%. Conclusions The RAPD PAGE fingerprinting is an useful method to type M tuberculosis strains. The prevalence of tuberculosis resisting to rifampin in Chongqing city is primarily due to the transmission of type Ⅰ, Ⅱ and Ⅲ strains.