肉类研究Issue(11)：34-37,4.
沙门氏致病菌标准阳性模板的构建及实时荧光定量聚合酶链式反应检测
Construction of Standard Positive Template and Development of a Real-Time Fluorescence Quantitative Polymerase Chain Reaction (FQ-PCR) Assay for PathogenicSalmonella spp.
摘要
Abstract
Objective: To construct recombinant plasmids for use as standard positive template and establish a real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) assay for determining pathogenicSalmonellaspp. in foods. Methods: Primers and Taqman probe were designed and synthesized with the specific fragment of InvA gene as the target sequence. Recombinant plasmids were constructed by inserting the target gene into PGM-T carriers. Real-time fluorescent quantitative PCR method was established and its sensitivity, specificity, repeatability and accuracy were investigated. Results: The recombinant plasmids constructed using the specific sequence of pathogenicSalmonella spp. could be used as a standard positive template for fluorescent quantitative PCR. The standard curve wasY=-3.151 lgX+42.86 (R2=0.999), and the sensitivity of the method was 80 copies per reaction. It was specific to detect Salmonellaspp. with good repeatability (the inter-batch and intra-batch coefficients of variation were both less than 5%). Conclusion: The FQ-PCR method allows qualitative and quantitative detection of pathogenicSalmonellaspp.关键词
沙门氏致病菌/标准阳性模板/实时荧光定量聚合酶链式反应/TaqMan探针Key words
Salmonella spp./standard positive template/fluorescent quantitative polymerase chain reaction (FQ-PCR)/TaqMan probes分类
生物科学引用本文复制引用
陈晨,邵彪,陈刚,黄伟东..沙门氏致病菌标准阳性模板的构建及实时荧光定量聚合酶链式反应检测[J].肉类研究,2014,(11):34-37,4.基金项目
2013年度国家星火计划项目（2013GA690155）；南通市科技计划项目 （）
