荧光定量PCR快速检测沙门菌方法的建立及应用

目的:建立沙门菌实时荧光PCR的快速检测方法,探讨其可行性和应用价值。方法:根据沙门菌的特异性DNA作为靶序列,以沙门菌菌株提取核酸DNA作为模板进行荧光检测。结果:本研究在7种相关菌株的检测中,除沙门菌出现很好的阳性外,其余菌株均为阴性;在纯菌条件下,定量检测低限为30 cfu/ml;同一样品检测3次Ct值的变异系数均小于5%;对模拟标本与分离培养对比,二者符合率为100%。结论:该方法的建立,不仅为食源性沙门菌污染及食物中毒的快速检测提供依据,而且还可直接用于临床粪便标本的检测。

Establishment and application of real-time fluorescence quantitative PCR for detection of Salmonella

Objective:To establish a real-time fluorescence quantitative PCR method for rapid detection of Salmonella and investigate the feasibility and application value.Methods: Based on Salmonella specific gene,DNA extracted from Salmonella strain was used as template for fluorescence detection.Results: Results of all 7 bacteria strains were negative except for strains of Salmonella.The detection limit of the method was 30 cfu/ml with pure cultures of Salmonella.Coefficient variables were all less than 5% in 3 different detections of the same sample.To compare simulation specimens with culture strains,coincidence rate was 100%.Conclusion: This method can provide the basis for rapid detection of food-borne contamination of Salmonella and direct clinical stool specimens.