PCR和DNP标记核酸探针检测禽源沙门氏菌四环素耐药性基因tetC的研究 (pdf)

    【摘要】 目的 观察利用PCR和DNP标记核酸探针检测禽源沙门氏菌四环素耐药性基因tetC。方法 通过对临床分离并经生化实验和血清学试验鉴定的11株禽源沙门氏菌和1株标准株C79-13进行药敏试验。结果 有11株有高耐药性，另外1株有低耐药性，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，与药敏试验符合率为100%；用DNP标记其PCR产物，制成DNA探针，采用菌落杂交方法对12株沙门氏菌菌落进行杂交。结果显示：与PCR结果的阳性符合率为90.2%，具有很高的特异性。结论 利用PCR和用DNP标记的这种探针检测四环素耐药基因tetC具有很高的敏感性和特异性，从而为禽类沙门氏菌的耐药性检测提供了高效敏感的手段。

    【关键词】 沙门氏菌；四环素耐药基因；TetC； PCR；核酸探针

    Study on detection of tetracycline resistance gene(tetC) of avian pathogenic Salmonella by PCR and DNP labeled nucleic acid probe

    ZHANG Zheng-Qing , LUO Wei , LIU Ya-Gang ,et al.College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041,China

    【Abstract】 Objective By using clinical isolates and biochemical and serologic tests detection on 11 avian Salmonella strains and one standard Salmonella C19-13, the drug-susceptibility test shows that: 11 strains are with high resistance, another 1 is low.Methods Results The drug-susceptibility tests also shows that the rate is 100%. After amplifying the tetracycline-resistance gene, it produces specific material by plasmids complete and the concordance rate to drug-susceptibility test is 100%. By using the method of probing the tetC gene labeled with DNP and use colony hybridization to 12 strains, the method of PRC showed 90.2% concordance in positive rate and with high speciality. Conclusion The result indicates that the probe of tetC gene with PCR to detect tetracycline-resistant gene has the character of susceptibility and speciality. The paper will provides effective and exact methods to detect the drug-resistance of avian Salmonella. ......