运用UP-PCR技术进行食源性感染常见致病菌Hsp60基因的扩增与酶切鉴定

The Amplification and Identification of Hsp60 Gene of Foodborne Pathogens with UP-PCR Method

目的 扩增、鉴定食源性感染常见致病菌Hsp60基因。 方法 选取 13种食源性感染常见致病菌 ,采用UP -PCR法扩增其Hsp60基因 ,并对PCR产物进行酶切鉴定分析。 结果 13种食源性感染常见致病菌均可以扩增出约 1.1KB的基因片段 ,蜡样芽孢杆菌PCR产物经HindIII酶切后形成约 684bp和 470bp片段 ,小肠结肠炎耶尔森菌PCR产物经BglII酶切后形成 861bp和 2 96bp片段。 结论 UP -PCR技术能同时扩增出食源性感染常见致病菌Hsp60基因 ,为进一步进行致病菌的种属鉴定和分型研究奠定基础。

Objective To amplify and identify Hsp60 gene fragment of foodborne pathogens. Methods Hsp60 gene fragment of thirteen representative foodborne pathogens were amplified with UP-PCR method. PCR products were purified and identified by restriction enzyme digestion. Results 1.1 Kbp gene fragment were amplified from all target pathogens. After being digested with restriction enzyme Hind III, 684 bp and 470 bp fragment were got from Bacillus cereus PCR product, and 861 bp and 296 bp fragment from Yersinia enterocolitica PCR product with restriction enzyme Bgl II digestion. Conclusion Hsp60 gene fragment of foodborne pathogens could be amplified with UP-PCR method, which has laid a good basis for further species and serotype identification of food-borne pathogens.