Establishment and evaluation of a real-time IAC-PCR for the detection of Salmonella
添加有扩增内标的沙门氏菌荧光定量PCR 检测体系的建立与评价

Objective] The aim of this study was to establish a new EvaGreen real-time IAC-PCR for the rapid detection of Salmonella.Methods] We used Salmonella genomic comparison analysis to mine Salmonella-specific targets,and Primer Premier 5.0 to design primers which were evaluated by specificity and sensitivity tests.Results] We obtained a Salmonella-specific gene that encodes putative type Ⅲ secretion protein(ssaQ),and specific primers(SsaQ6L/SsaQ6R) were designed based on this gene.Then we established IAC-PCR and EvaGreen real-time IAC-PCR assays,which showed 100% inclusivity and 100% exclusivity on all strains tested.Their detection limits of purified Salmonella genomic DNA were 14.9 copies/PCR and 2.76 copies/PCR respectively.Artificial contamination assays showed that Salmonella could be detected after 10 hours and 8 hours enrichment when the original bacterial concentration was 4.2 cfu/10 mL.Conclusion] A new EvaGreen real-time IAC-PCR with high specificity and sensitivity was successfully developed for the rapid detection of Salmonella.