摘要
为建立一种快速、灵敏、特异的实时荧光定量PCR法用于沙门菌的检验,根据GenBank上登录的编号为AE016841的沙门菌序列,应用生物学软件在fimY基因的保守区设计引物和TaqMan探针,同时应用BLAST程序进行网上序列比对,并进行筛选、优化。用鼠伤寒标准菌和60份食品样本进行本检测方法的特异性、敏感性和重复性试验,并与常规法和科玛嘉平板分离法做比较。本方法对沙门菌的检测具高度的特异性,检测的灵敏度达102CFUml,从增菌至完成检测仅需24h左右,是一种快速检测沙门菌的敏感、特异的新方法。
To establish a rapid, sensitive and specific method of TaqMan-based real-time PCR assay for detection of salmonella, based on the gene sequence of salmonella from GenBank （ accession no, AE016841 ）, the specific primers and probes were designed in the conserved regions of the fimY gene by using the biologic software. The specificity of the sequences were tested by BLAST search in GenBank. This assay was applied to the S. typhimurium and 60 food samples to test its specificity, sensitivity and repeatability, compared with those of the regular bacterial culture and the culture on CH ROMager plate. The assay was proved to be highly specific to salmonella, the sensitivity was 102 CFU/ml, and it only took 24 hours to complete the whole assay. It is concluded that the TaqMan-based real-time PCR assay is a rapid, sensitive and specific method for the detection of salmonella.
出处
《中国食品卫生杂志》
2006年第4期311-313,共3页
Chinese Journal of Food Hygiene
