%0 Journal Article %T 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 %A Xianlong Dan %A Bin Liu %A Lida Zhang %A Xiaoling Li %A Chunlei Shi %A Min Zhou %A Xianming Shi %A
但现龙 %A 刘斌 %A 李小玲 %A 张利达 %A 施春雷 %A 周敏 %A 史贤明 %J 微生物学报 %D 2011 %I %X 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. %K Salmonella %K Target mining %K EvaGreen real-time PCR %K Internal amplification control(IAC)
沙门氏菌 %K 靶点筛选 %K EvaGreen荧光定量PCR %K 扩增内标 %U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=EEC77E274DE7318A9B8735C21460ECA3&yid=9377ED8094509821&vid=987EDA49D8A7A635&iid=5D311CA918CA9A03&sid=2625CEFACC964DE3&eid=FB318F788ECEF1B2&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=28