摘要
目的:建立沙门菌实时荧光定量PCR快速检测方法,探讨沙门菌在食源性感染快速诊断中的应用价值。方法:采用沙门菌fimY基因序列设计特异引物和探针,建立实时荧光实时定量PCR技术检测沙门菌的检测方法。通过特异性、敏感性、稳定性和重复性,以验证方法的可行性。结果:成功构建了沙门菌重组质粒标准品,为方法学建立奠定基础;通过特异性、敏感性、稳定性和重复性验证,结果表明具有较好的特异性,最低检测限为103拷贝/ml,相当于100 cfu/ml的菌细胞,并有良好的稳定性和重复性。结论:沙门菌实时荧光定量PCR检测方法的建立,不仅为食源性沙门菌污染的快速检测提供依据,而且还可直接用于临床粪便标本检测。
Objective：To establish a real-time fluorescence quantitative PCR method for rapid detection of Salmonella,and to explore food-borne Salmonella infection in rapid diagnosis.Methods：Primer and probe were designed based on femB gene sequence of Salmonella,to establish real-time fluorescence quantitative RT-PCR detection method for Salmonella.Through specificity,sensitivity,stability and repeatability,to verify the feasibility of the method.Results：Standard reconstructed plasmid and Salmonella-specific fluorescence quantitative RT-PCR method were established successfully.After laboratory evaluation on stability,specificity,repeatability and sensitivity.The lowest limitation for detecting Salmonella by real-time fluorescence quantitative PCR was 10^3 copy/ml,equivalent to the bacteria cells 100 cfu/ml.Conclusion：The establishment of real-time fluorescent quantitative RT-PCR methods specific for Salmonella will provide the basis for rapid detect food-borne contamination of Salmonella,also direct detect clinical stool specimens.
出处
《中国卫生检验杂志》
CAS
2010年第6期1302-1304,共3页
Chinese Journal of Health Laboratory Technology
关键词
沙门菌
实时荧光定量PCR
食源性检测
Salmonella
Real-time fluorescence quantitative PCR
Detection of food-borne
