PCR-RFLP和PCR-SSCP在食源性感染致病菌检验中的应用

摘要

目的 探讨PCR-RFLP和PCR-SSCP在食源性感染致病菌检验中的应用.方法 将鉴别靶基因设定为食源性感染14种常见致病菌的23S rRNA基因,扩增时运用通用引物PCR(UP-PCR),并运用单链构象多态性分析(SSCP)和限制性酶切片段长度多态性分析(RFLP)分析PCR产物的酶切片段.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的RFLP图谱相似;SSCP分析显示,SSCP图谱具有较大的变异性,能够为细菌鉴定提供有效依据.结论 PCR-SSCP技术能够快速鉴定食源性感染致病菌,值得在临床推广应用.%Objective To investigate the application of PCR-RFLP and PCR-SSCP in the inspection of foodborne pathogens.Methods 23S rRNA gene of 14 kinds of common foodborne pathogens were set as target genes,universal primers PCR (UP-PCR) was used in amplification,and restriction fragment length polymorphism (RFLP) analysis and single strand conformation polymorphism (SSCP) analysis were used to analyze PCR products digested fragments.Results RFLP for Hpa Ⅱ digestion products 23S rRNA gene fragment analysis showed that RFLP pattern of different species of bacteria had similar performance;SSCP analysis showed that SSCP patterns had greater variability,so could provide an effective basis for the identification of bacteria.Conclusion PCR-SSCP technique can rapidly identify foodbome pathogens,so is worthy of promotion and application in the clinical.