摘要
目的探讨运用PCR-RFLP和PCR-SSCP技术建立肠杆菌科食源性感染常见致病菌快速鉴定方法的可行性。方法选取肠杆菌科食源性感染14种常见致病菌的23 S rRNA基因作为鉴别靶基因,采用通用引物PCR(UP-PCR)进行扩增,并对PCR产物的酶切片段进行限制性酶切片段长度多态性分析(RFLP)和单链构象多态性分析(SSCP)。结果针对23S rRNA基因片段的Hpa II酶切产物的RFLP分析表明,不同种属的致病菌表现出相似的RFLP图谱。SSCP分析显示,14种肠杆菌科常见致病菌SSCP图谱变异性很大,有利于进行细菌鉴定。结论运用PCR-SSCP技术可进行肠杆菌科食源性感染致病菌的快速鉴定。
Objective To study the feasibility of establishment a method for the identification of Enterobacteriaceae foodborne infectious bacteria with polymerase chain reaction-restricted fragment length polymorphism （PCR- RFLP） and PCR - single strand conformation polymorphism （PCR - SSCP） techniques. Methods By using the universal primer PCR, the 23S rRNAs from the 14 bacteria cultures were amplified. The PCR products were analyzed by RFLP and SSCP and evaluated respectively. Results The RFLP results of endonuclease Hpa Ⅱ digested product of the 23S gene rRNAs of the different Enterobacteriaceae bacteria manifested in similar patterns, but the SSCP analysis showed differential patterns among the 14 Enterobacteriaceae bacteria species and were valid for identification of the target bacteria. Conclusion PCR-SSCP can rapidly identify Enterobacteriaceae foodborne infection bacteria.
出处
《实用预防医学》
CAS
2005年第6期1263-1265,共3页
Practical Preventive Medicine
基金
广东省自然科学基金资助项目(53201420202850111)
