致病性猪沙门菌四环素耐药基因tetB的PCR检测  

PCR amplification of tetB gene for pathogenic Salmonella isolated from diseased pigs

作　　者：刘维红[1] 徐引弟[1] 郭爱珍[1] 贾爱卿[1] 刘军发[1] 陈焕春[1] 

机构地区：[1]华中农业大学农业微生物学国家重点实验室,武汉430070

出　　处：《中国抗生素杂志》2007年第1期51-55,共5页Chinese Journal of Antibiotics

基　　金：国家自然科学基金资助项目(30571386);中国教育部留学回国人员科研基金(教外司留[2004]176号)

摘　　要：目的检测临床分离猪源致病性沙门菌四环素的耐药基因,确定沙门菌四环素耐药基因类型。方法用KB法测定分离株对四环素等21种抗生素的耐药情况,用PCR方法检测四环素耐药基因tetA、tetB、tetC、tetD、tetE、tetG及tetK,并对扩增产物进行测序。结果临床分离株全部对四环素耐药,33株沙门菌中有29株扩增出tetB特异性片段,基因序列与参考菌的同源性为99%,tetB检测与药敏试验阳性符合率为87.9%。结论tetB的PCR检测对四环素耐药性具有较高的特异性,tetB基因是决定本试验中临床分离株四环素耐药的主要基因,为四环素耐药性的分子流行病学监测提供了依据。Objective To detect the tetracycline resistance genes for 33 Salmonella isolates from diseased pigs and define the tetracycline resistance determinants. Methods The resistance of Salmonella isolates were detected by drug susceptibility tests with Kirby-Bauer （KB） diffusion method. PCR was designed to detect tetA, tetB, tetC, tetD, tetE, tetG and tetK. The amplified products were sequenced and alignment analysis was performed. Results The detection of tetracycline-resistance of 33 Salmonella isolates showed the resistance rate was 100%, but only the tetB was amplified from 29 of 33 isolates. The gene fragment shared 99% homology in nucleotides with the reference sequence. The result of PCR amplification was 87. 9R% identical with that of the drug resistance test. Conclusion The tetB was the main determinant responsible for tetracycline resistance in this＇study and PCR detection of tetB is a specific and reliable method for molecular epidemiology of tetracycline resistance in Salmonella.

关 键 词：沙门菌 四环素耐药基因 tetB PCR 猪 

分 类 号：R378.22[医药卫生—病原生物学]