荧光定量RT-PCR检测沙门氏菌方法的建立  

Real-time fluorescence quantitative PCR detection of Salmonella in experimental study

作　　者：杨柳[1] 苏明权[1] 马越云[1] 焦刚 常亮[1] 肖凤静[1] 李明 彭年才 郝晓柯[1] 

机构地区：[1]第四军医大学西京医院全军临床检验医学中心,陕西西安710032 [2]西安天隆科技有限公司,陕西西安710043

出　　处：《中国实验诊断学》2011年第1期17-20,共4页Chinese Journal of Laboratory Diagnosis

摘　　要：目的建立沙门氏菌实时荧光定量PCR的快速检测方法 ,探讨该方法的可行性和应用价值。方法根据沙门氏菌fimY基因序列设计引物和探针,采用基因重组技术构建用于沙门氏菌检测的定量标准品,建立实时荧光定量PCR检测沙门氏菌的方法。结果成功构建了沙门氏菌重组质粒标准品和沙门氏菌实时荧光定量PCR方法 ;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、敏感性、稳定性和重复性;对模拟标本与分离培养对比,二者符合率为100%。结论沙门氏菌实时荧光定量PCR检测方法的建立,为食源性沙门氏菌污染的快速检测提供依据,可应用于食品卫生监管、商品检验检疫以及临床诊断等。Objective To establish real-time fluorescent quantitative PCR methods for rapid detecting Salmonella and investigate the feasibility and application of value.Methods Primer and probe were designed based on fimY gene sequence of Salmonella,quantitative standards for Salmonella were set up with molecular recombination methods,and established real-time fluorescence quantitative PCR for Salmonella.Results Standard of reconstructed plasmid and real-time fluorescence quantitative PCR method were established successfully.It was show that this method has good specificity,sensitivity,stability by the verification experiment.To compared simulation specimens with culture strains,coincidence rate was 100%.Conclusion The establishment of real-time fluorescent quantitative PCR detection of Salmonella provide the basis for rapid detect food-borne contamination of Salmonella.It can be applied to food hygiene regulation,quarantine and clinical diagnosis.

关 键 词：沙门氏菌 实时荧光定量PCR 

分 类 号：R440[医药卫生—诊断学]