奶牛结核分枝杆菌rpoB和katG基因突变与多重耐药的相关性  

Cows Mycobacterium tuberculosis rpoB and katG Gene Mutations Associated with Multi-drug Resistance

作　　者：赵莉[1] 纳玮[1] 周学章[1] 王玉炯[1] 

机构地区：[1]宁夏大学西部特色生物资源保护与利用教育部重点实验室,宁夏银川750021

出　　处：《中国畜牧兽医》2011年第1期47-50,共4页China Animal Husbandry & Veterinary Medicine

基　　金：国家重点基础研究发展计划"973"项目(2006CB504400);高等学校科技创新工程重大项目(706057)

摘　　要：本试验旨在研究奶牛结核分枝杆菌rpoB和katG基因突变与耐药性之间的关系。将临床分离的30株牛源结核分枝杆菌,利用噬菌体生物扩增法(phage amplified biologically assay,PhaB)检测其对利福平(RFP)和异烟肼(INH)的药物敏感性,采用聚合酶链式反应—DNA直接测序法(PCR-DS)扩增rpoB和katG基因并测序,分析rpoB和katG基因突变及其耐药相关性。15株表型耐药菌株中4株单耐RFP,PCR产物直接测序后发现3株在rpoB基因531位点发生突变,突变率为75.00%(3/4);9株单耐INH菌株中7株发生突变,6株katG基因315位点基因突变,突变率为85.71%(6/7),1株发现463位点密码子基因突变,突变率为14.29%(1/7);2株耐RFP的同时也对INH耐药,测序发现2株均在rpoB基因的531位点和katG基因的315位点发生突变。所有菌株均未发现rpoB和katG基因的缺失。rpoB基因531位点和katG基因315位点的突变是RFP和INH产生耐药性的主要机制;检测牛源结核分枝杆菌RFP耐药性的同时可以初步筛选耐多药结核分枝杆菌。The study of cows Mycobacterium tuberculosis rpoB and katG gene mutations and the relation between drug resistance.Take 30 strains of clinical isolates of M.tuberculosis from cattle,use of phage amplified biologically assay on the drug sensitivity of Rifampicin and isoniazid,polymerase chain reaction-DNA direct sequencing rpoB and katG gene were amplified and sequenced,analysis of rpoB and katG gene mutations and drug resistance associate.The results showed that 15 resistant strains in four were resistant to RFP,three of them found mutations at condon 531 occurred in 75.00%（3/4）;9 single INH-resistant strains of seven mutations,6 isolates mutation at condon 315 occurred in 85.71%（6/7）,1 isolate mutation at condon 463 occurred in 14.29%（1/7）;2 isolates were resistant to RFP and INH,sequencing showed that two were both mutations at condon 531 in the rpoB gene and condon 315 in the katG gene.All strains were not found the deletion of rpoB and katG gene.The main mechanism of RFP and INH drug resistance is mutations at condon 531 in the rpoB gene and condon 315 in the katG gene;while detection of bovine tuberculosis RFP-resistance,it can be initial screening MDR M.tuberculosis.

关 键 词：结核分枝杆菌 利福平 异烟肼 RPOB基因 KATG基因 多重耐药 

分 类 号：S855.2[农业科学—临床兽医学]