摘要
根据沙门菌保守的fimY基因序列设计合成了引物和TaqMan探针,建立了快速检测沙门茵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。通过对临床样品的检测,该试剂盒的检测灵敏度达4．5 CFU(25μL反应体系),比常规PCR高100倍,而且稳定性良好,至少可以冷冻保存9个月。结果表明,所建立的沙门菌实时荧光定量PCR检测试剂盒具有操作简便、快速、特异性强、灵敏度高、重复性好、稳定性强等优点,适于大量样品的检测。
The Taqman probe and primers were designed and synthesized according to the conserved gene fimY of Salmonella available in GenBank, and then reaction parameters were optimized to develop a real time Taqman quantitative PCR detection kit. The developed quantitative PCR kit could detect 4.5 CFU in the 25 μL-reaction-system. Its sensitivity was 100 times higher than that of the routine PCR, and it could be stored over nine months at -20 ℃. It was confirmed through detection of some clinic samples that the developed kit was the simple, sensitive, specific, accurate, good repeated and stable method for the rapid detection of Salmonella and could be applied to detection of a large number of samples.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2006年第8期625-629,共5页
Chinese Veterinary Science
基金
厦门市科技计划项目(3502Z20042021)
