PCR和核酸探针检测猪源沙门氏菌四环素耐药基因tetC的研究  

Research on Detection of Tetracycline Resistance Gene(tetC) of Pathogenic Salmonellafrom Swine by PCR and Nucleic Acid Probe

作　　者：代敏[1] 王红宁[1] 吴琦[1] 

机构地区：[1]四川农业大学动物科技学院

出　　处：《畜牧兽医学报》2005年第5期482-485,共4页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家"十五"科技攻关课题(2002BA514A 17 11)

摘　　要：对沙门氏菌的四环素耐药性进行了检测,结果表明,沙门氏菌对四环素类抗生素产生了广泛的耐药性,耐药率达100%,对其四环素耐药基因tetC进行了扩增,结果获得以质粒为模板的特异性产物,与药敏试验结果阳性符合率75%,具有较高的检出率。用光生物素对PCR产物进行标记,制备核酸探针,采用菌落原位杂交的方法对沙门氏菌进行检测,结果表明13株阳性、3株阴性,杂交结果与PCR结果阳性符合率为93.75%,具有较高的特异性。通过条件的优化,建立的四环素耐药基因tetC的PCR和核酸探针检测技术,为四环素耐药性的分子流行病学监测提供了有效的途径。The detection of tetracyclines-resistance of 16 Samonella isolates showed the resistance rate was 100%. The tetC gene of 16 salmonella strains was amplified by polymerase chain reaction. 12 strains with this gene were located on plasmids. Compared with the result of the drug-susceptibility tests, the method of PCR showed 75% concordance in positive rate. The probe of tetC gene was labelled with photobiotin,and 16 pathogenic Salmonella strains from swine were detected by colony hybridization in situ. The result indicated that 13 strains were positive, 3 strains were negative. Compared with the result of PCR, the method of the tetC probe showed 93.75% concordance in positive rate. In this study, two methods for detecting tetC gene were established by using PCR and nucleic acid probe, and they could be applied to the molecular epidemiology surveillance of tetracyclines-resistance.

关 键 词：沙门氏菌 TETC 耐药基因 核酸探针 PCR 四环素类抗生素 流行病学监测 PCR产物 耐药性 试验结果 原位杂交 检测技术 特异性 符合率 猪 耐药率 阳性 检出率 生物素 药敏 

分 类 号：S858.315.1[农业科学—临床兽医学] S859.796[农业科学—兽医学]