摘要
根据GenBank公布的单增李斯特菌的hlyA基因序列和金黄色葡萄球菌的nuc基因序列各设计一对引物和一条探针,建立基于TaqMan探针的单增李斯特菌和金黄色葡萄球菌双重荧光定量聚合酶链式反应(PCR)检测方法。该方法对所有目标菌株均产生特异性扩增曲线,其他非目标菌均不产生扩增曲线,具有较强的特异性。且该法对阳性重组质粒pMD18-hlyA和pMD18-nuc同时定量扩增的敏感度分别为19.5拷贝/μL和18.7拷贝/μL。同步检测人工染菌肉样中单增李斯特菌和金黄色葡萄球菌的最低检出限均为102CFU/g。
A TaqMan duplex fluorescence quantitative PCR assay for detecting Listeria monocytogenes and Staphylococcus aureus was presented.Specific primers and probes were designed according to the hlyA gene sequence of Listeria monocytogenes and the nuc gene of Staphylococcus aureus published in GenBank.The developed assays detected only the two pathogens,respectively,but not other bacteria,indicating the high specificity of the duplex PCR.When the positive recombinant plasmids pMD18-hlyA and pMD18-nuc were simultaneously amplified,the sensitivity for them was 19.5 copy/μL and 18.7 copy/μL,respectively,and the limit of detection was 102 CFU/g for both Listeriamonocytogenes and Staphylococcus aureus in artificially inoculated meat.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2013年第16期169-172,共4页
Food Science
基金
"十二五"国家科技支撑计划项目(2012BAD28B02)
