结核分枝杆菌对氨基水杨酸耐药相关基因的筛选及鉴定  

Screening and identification of the PAS resistance related genes

作　　者：郑晓静[1,2] 杜博平 贾红彦[1,2] 侯继增 杨超 高汉青 杜凤娇[1,2] 邢爱英[1,2] 李自慧[1,2] 曹廷明[1,2] 张宗德[1,2] 李琦[4,5] 

机构地区：[1]首都医科大学附属北京胸科医院 [2]北京市结核病胸部肿瘤研究所结核病分子生物学研究室,101149 [3]北京市通州区结核病防治所检验科 [4]首都医科大学附属北京胸科医院结核三科 [5]北京市结核病胸部肿瘤研究所

出　　处：《北京医学》2012年第9期783-786,共4页Beijing Medical Journal

基　　金："艾滋病和病毒性肝炎等重大传染病防治" 科技重大专项(2012ZX10003002-003);北京市卫生系统高层次卫生技术人才培养计划(2009-3-51)

摘　　要：目的筛选二线抗结核药物对氨基水杨酸(para-aminosalicylic acid,PAS)的耐药相关基因。方法构建H37Rv转座子文库,筛选PAS耐药克隆,随机引物PCR扩增转座子插入突变基因。在PAS耐药菌株中鉴定耐药相关基因的突变情况。结果构建了较完整的H37Rv转座子文库,并筛选出201个PAS耐药克隆,经测序找到4个可能的PAS耐药相关基因,分别是Rv1534、pknF、ndh、papA1。papA1基因检测到一个无义突变,为papA1基因339位C→T点突变。结论酰基转移酶基因papA1可能为PAS耐药相关基因。Objective To screen the PAS （para-aminosalicylic acid） resistant-related genes. Methods The H37Rv transposon-insertion library was constructed. The PAS resistant clone was screened with random primer PCR, and mutant gene with transposon-insertion was identified. Furthermore, the mutants of the PAS resistant-related genes were examined in the PAS clinical resistant strain. Results The H37Rv transposon-insertion library was constructed successfully, and 4 probably PAS resistant-related genes （Rv1534,pknF.ndh.papA1） were identified. A nonsense mutation of papa 1 was found in the PAS clinical resistant strain. Conclusion papA 1 is probably the PAS resistant-related gene.

关 键 词：结核分枝杆菌 耐药 对氨基水杨酸 基因 

分 类 号：R440[医药卫生—诊断学]