Construction of Standard Positive Template and Development of a Real-Time Fluorescence Quantitative Polymerase Chain Reaction (FQ-PCR) Assay for PathogenicSalmonella spp.
CHEN Chen;SHAO Biao;CHEN Gang;HUANG Weidong
1. Nantong Products Quality Supervision and Inspection Institute, Nantong 226011, China;
2. School of Life Science and Technology, China Pharmaceutical University, Nanjing 211198, China
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.
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, 28(11): 34-37.
CHEN Chen;SHAO Biao;CHEN Gang;HUANG Weidong. Construction of Standard Positive Template and Development of a Real-Time Fluorescence Quantitative Polymerase Chain Reaction (FQ-PCR) Assay for PathogenicSalmonella spp.. Meat Research, 2014, 28(11): 34-37.
2014, Vol. 28 
