摘要
[目的]建立一种快速、灵敏、准确的检测方法,预防沙门菌食物中毒。[方法]根据沙门菌保守基因序列fimY基因设计合成了引物和TaqMan探针,建立快速检测沙门菌的实时荧光定量方法,并对反应条件进行了优化,组装成快速检测试剂盒。[结果]其检测灵敏度达到4．5efu,比常规PER高100倍,而且稳定性良好,冷冻至少可以保存9个月。[结论]通过临床样品的检测,该试剂盒具有操作简便快速、灵敏度高、特异性强、重复性好、稳定性强等优点,很适合大量样品的检测。
Objective To establish a simple, sensitive, accurate and rapid detection method screening salmonella in food.Method The Taqman probe and primers were designed and sythesized according to the conserved gene fimY of salmonella available in GenBank, and then reac- tion parameters were optimized to develop a real time Taqman quantitative PCR detection kit. Result The developed quantitative PCR kit could detect 4.5cfu/25uL（a reaction system）, its sensitivity was 100 times higher than that of the routine PCR, and it could be stored over nine months in - 20℃. Conclusion The developed kit has many advantages in detecting some clinic samples, such as simple, sensitive, specific, accurate, easy to duplication stable for rapid detection in salmonella and is apphed to a large number detection.
出处
《中国国境卫生检疫杂志》
CAS
2006年第B08期111-114,共4页
Chinese Journal of Frontier Health and Quarantine
关键词
沙门菌
探针
实时荧光定量
PCR
检测
Salmonella
TaqMan probe
Real- time fluorescence quantitative PCR
Detection
