肉类研究2017，Vol.31Issue(2)：45-50,6.DOI:10.7506/rlyj1001-8123-201702009
低温肉制品中3种食源性致病菌多重聚合酶链式反应快速检测方法的建立
Multiplex Polymerase Chain Reaction Assay for Rapid Detection of Three Foodborne Pathogens in Low-Temperature Meat Products
摘要
Abstract
Salmonella spp.,Listeria monocytogenes (L.monocytogenes) and Staphylococcus aureus (S.aureus) are 3 species of bacteria which can easily contaminate low-temperature meat products.The objective of this study was to develop a rapid and efficient method for the simultaneous detection of the bacterial pathogens using multiplex polymerase chain reaction (m-PCR).The specific primers for the invA gene of Salmonella spp.,the PrfA gene ofL.monocytogenes and the nuc gene of S.aureus were designed and their specificity was determined.After enrichment culture,the 3 strains were inoculated to lowtemperature meat products and incubated overnight for evaluating the sensitivity of the detection method.Optimization of the PCR system and its application to the detection of real samples were carried ont.Our experimental results showed that the optimum PCR conditions were determined as follows:using a reaction system containing 2.5 μL of 10 × PCR buffer,2.0 μL of 2.5 mmol/L dNTPs,0.5 μL of 2.5 U/μL DNA polymerase,0.5 μL of upstream primer,0.5 μL of downstream primer,1.0 μL of L.monocytogenes template,1.0 μL of S.aureus template or 2.0 μL of Salmonella spp.template brought with ddH2O up to 25 μL,and an annealing temperature of 64 ℃.The multiplex PCR method was applicable to detect Salmonella spp.in meat products with the advantages of rapidity,high sensitivity and specificity and simplicity.关键词
低温肉制品/多重PCR/快速检测/食源性致病菌Key words
low-temperature meat products/multiplex polymerase chain reaction (m-PCR)/rapid detection/foodborne pathogens分类
轻工纺织引用本文复制引用
李新福,王周平,李聪,方伶俐,赵颖,徐宝才..低温肉制品中3种食源性致病菌多重聚合酶链式反应快速检测方法的建立[J].肉类研究,2017,31(2):45-50,6.基金项目
国家自然科学基金面上项目(31571909) （31571909）
“十三五”国家重点研发计划重点项目(2016YFD0400703-05) （2016YFD0400703-05）
