PCR-RFLP和PCR-SSCP技术对常见食源性感染致病菌23S rDNA基因的鉴定

Application of PCR-RFLP and PCR-SSCP technique for identification of 23 S rDNA gene of foodborne pathogenic bacteria

目的 探讨应用PCR RFLP和PCR SSCP进行食源性感染常见致病菌鉴定的可行性。方法 选择细菌 2 3SrDNA基因作为靶基因 ,设计合适的通用引物进行PCR扩增 ,对 13种食源性感染常见致病菌扩增产物的酶切片段进行RFLP和SSCP分析。结果 通用引物可以扩增出 13种食源性感染常见致病菌的 2 3SrDNA基因 ,HpaⅡ酶切产物的RFLP分析表明 ,不同种属的致病菌表现出不同的RFLP图谱 ;SSCP分析表明 ,13种食源性感染常见致病菌SSCP图谱变异性更大 ,不利于种属间鉴别 ,但有利于种内的鉴定。结论 运用PCR RFLP和PCR

Objective To study the feasibility of PCR-RFLP and PCR-SSCP technique in analysis of foodborne pathogenic bacteria.Methods 23 S rDNA gene was selected as target fragment and universal oligonucleotide primers were designed and synthesized.PCR product of 13 species from foodborne pathogenic bacteria were enzymolyzed with restriction enzyme Hpa Ⅱ and subsequent PCR-RFLP and PCR-SSCP were performed.Results 23 S rDNA gene fragment was amplified from all the 13 species of foodborne pathogenic bacteria.RFLP analysis of enzyme product manifested that different species bacteria have different RFLP enzymogram.With respect to SSCP analysis much more mutante fragment displayed in different species of foodborne pathogenic bacteria and it might be beneficial to subspecies discrimination,but not beneficial to genra/species discrimination.Conclusion PCR-RFLP and PCR-SSCP could be used in the discrimination diagnosis of foodborne pathogenic bacteria.