%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