牛结核分支杆菌临床分离株rpsL,rpoB,KatG基因突变分析

Mutation Analysis on rpsL,rpoB,KatG Gene of Clinical Isolates Mycobacterium bovis

采用噬菌体生物扩增法对本实验室分离鉴定的25株奶牛结核分支杆菌进行药敏分析,检测出14株耐链霉素、利福平、异烟肼菌株.用PCR扩增耐药株的rpsL基因片段(370 bp)、rpoB基因片段(213 bp)和KatG基因片段(458 bp),并对目的片段进行测序分析,其中5株耐链霉素菌株的rpsL基因在43位点发生突变,均由赖氨酸密码子(AAG)突变为精氨酸密码子(AGG);5株利福平耐药株的rpoB基因在531位点、526位点、511位点发生突变,其中有3株分离株的rpoB基因在531位点发生点突变,1株分离株的rpoB基因在526位点发生突变,1株分离株的rpoB基因在531位点和511位点发生了较为少见的联合突变;4株异烟肼耐药菌株,其中2株在315位点发生碱基突变,2株在463位点发生碱基突变.

Drug susceptibility analysis of the 25 Mycobacterium boris which was isolated and identified in the laboratory was hold by phage biological amplified assay. 14 streptomycin, rifampicin, isoniazid-resistant strains was detected. The resistant strains of the rpsL gene fragment （370 bp）, rpoB gene fragment （213 bp） and KatG gene fragment （458 bp） was amplified by PCR, and the sequencing fragments of purpose was analyzed. There＇re 5 streptomycin-resistant strains＇ rpsL gene mutate in 43 site ,they＇ve all changed from lysine codon （AAG） to arginine codon （AGG） ; there＇re 5 rifampicin-resistant strains＇ rpoB gene mutate in 531, 526, 511 sites, three of which mutate in 531 site, one of which mutate in 526 site, the other mutate jointly in 531 site and 511 site which rarely occur; there＇re 4 isoniazid-resistant strains,two of which mutate in 315 site, the others mutate in 463 site.