结核分枝杆菌常见突变位点的克隆及应用  

Mycobacterium tuberculosis common mutant clones and application site

作　　者：马苗苗[1] 钟春蕾[1] 石亚萍[1] 彭文锋[1] 刘倩倩[1] 刘梅[1] 李娜娜[1] 林伟[1] 陈玲[1] 

机构地区：[1]遵义医学院附属医院呼吸二科,呼吸医学研究室,遵义贵州563003

出　　处：《临床医药文献电子杂志》2014年第13期1478-1479,共2页Electronic Journal of Clinical Medical Literature

基　　金：贵州省省长资金临床应用课题专项研究[黔省专合字(2012)123];贵州省结核病综合防治2011协同创新中心(培育)项目

摘　　要：目的克隆结核分枝杆菌rpoB、rpoC、katG、inhA基因突变位点,为核酸薄膜导流杂交技术快速检测提供阳性对照。方法将结核分枝杆菌基因12个单突变位点,包括rpo B基因526、516、511、522、550、572、1001位点,rpoC基因483位点,KatG基因315位点及inhA基因5、8、15位点进行TA克隆。结果成功克隆了rpo B、rpoC、katG、inhA基因上述12个单突变位点片段。结论结核分枝杆菌基因常见突变位点的成功克隆,为核酸薄膜导流杂交技术快速检测临床标本提供了阳性对照,有助于判断临床标本有无结核分枝杆菌感染并可提示有无耐药。Objectives To clone high frequent mutations of rpoB, rpoC, kat G and inh A genes in Mycobacterium Tuberculosis for positive controls in rapid detection of Drug-resistant Mycobacterium Tuberculosis isolates with oligonucleotide membrance-based flow-through hybridization technology. Methods A total of 12 mutation points which include 526,516,511,522,550,572 and 1001 codons of the rpoB gene, codon 483 of the rpo C gene, codon 315 of the kat G gene and codons 5,8,15 of the inhA gene were cloned into pMD19-T Vector. Results The 12 mutation genes of rpoB, rpoC, katG and inh A were successfully cloned. Conclusions cloning of the high frequent mutations in rpoB, rpoC, kat G and inh A genes could be applied in oligonucleotide arrays as positive control to detect mycobacterium tuberculosis infection and identify the drug resistance of the strains.

关 键 词：结核分枝杆菌 突变位点 克隆 

分 类 号：R378.911[医药卫生—病原生物学]