沙门菌；_四环素耐药基因；_tetB；_PCR；_猪

致病性猪沙门菌四环素耐药基因tetB的PCR检测

http://www.100md.com 《中国抗生素杂志》 2007年第1期

沙门菌；,四环素耐药基因；,tetB；,PCR；,猪,,沙门菌；,四环素耐药基因；,tetB；,PCR；,猪,1材料与方法,2结果与分析,3讨论,参考文献

     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。

    关键词：沙门菌；四环素耐药基因； tetB； PCR；猪

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

    Liu Weihong, Xu Yindi, Guo Aizhen, Jia Aiqing,Liu Junfa and Chen Huanchun

    (State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070)

    ABSTRACT 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9% 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. ......

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