山东泰安地区致病性猪沙门菌氨基糖苷类耐药基因aph(3)′-Ⅱa的PCR扩增与序列分析  

PCR amplification and nucleotide sequences analysis of aph(3′)-Ⅱa gene of pathogenic Salmonella from swine in Taian area

作　　者：王俊红[1] 牛钟相[1] 张文娟[1] 李晓静[1] 张永宁[1] 裴宗飞[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018

出　　处：《中国抗生素杂志》2008年第9期557-560,共4页Chinese Journal of Antibiotics

基　　金：山东省财政支持项目(SDGP-2004-54-0)

摘　　要：目的研究沙门菌对氨基糖苷类抗生素的耐药性并分析其耐药基因aph(3′)-Ⅱa的序列。方法对19株致病性猪沙门菌的氨基糖苷类抗生素耐药性进行检测;对其质粒上的aph(3′)-Ⅱa基因进行PCR扩增,选取其中任意一株(WWS171)的PCR阳性产物进行耐药基因aph(3′)-Ⅱa的序列分析。结果沙门菌对氨基糖苷类抗生素产生了广泛的耐药性,耐药率达100%;耐药基因aph(3′)-Ⅱa经PCR扩增后在13株沙门菌的质粒上出现582bp的特异性产物,与药敏试验结果的阳性符合率为68.4%,具有较高的检出率;与GenBank上发表的AF078924.1、AF188331.1、AY333434.1、AY598820.1、DQ842000.1的耐药基因aph(3′)-Ⅱa的序列完全相同。结论aph(3′)-Ⅱa的PCR检测对氨基糖苷类抗生素耐药性具有较高的特异性,aph(3′)-Ⅱa基因是决定本试验中临床分离株氨基糖苷类抗生素耐药的主要基因,为氨基糖苷类抗生素耐药性的分子流行病学监测提供了依据。Objective To study the aminoglycosides-resistance condition and aph （3′）-Ⅱa gene sequences of Salmonella. Methods Susceptibility tests of aminoglycosides-resistance were performed in 19 pathogenic Salmonella from swine. The aph（3′）-Ⅱa gene of 19 Salmonella strains was amplified by polymerase chain reaction. Choosing Salmonella WWS171 strain, which was one of the PCR positive products, its aph （3′）-Ⅱa gene sequences were analyzed. Results Nineteen Salmonella isolates showed highly resistance against aminoglycosides, the resistance rates was 100%. With PCR anplification, there was a 582bp fragment in the plasmids of 13 strains. Compared to the results of the drug-susceptibility tests, the method of PCR showed 68.4% concordance on positive rate. Detected aph（3′）-Ⅱa gene of Salmonella WWS171 strain had the same gene sequences as the genes （AF078924. 1, AF188331.1, AY333434. 1, AY598820. 1, DQ842000. 1） published in GenBank. Conclusions The aph（3′）-Ⅱa was the main determinant responsible for aminoglycosides-resis- tance in this study, and PCR detection of aph（3′）-Ⅱa was a specific and reliable method for molecular epidemiology of aminoglycosides resistance in Salmonella.

关 键 词：沙门菌 氨基糖苷类抗生素耐药基因 aph(3′)-Ⅱa PCR 序列分析 

分 类 号：R378.22[医药卫生—病原生物学]