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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。
Objective: To establish a rapid detection method of Salmonella real-time fluorescence PCR and to explore its feasibility and application value. Methods: According to the specific DNA of Salmonella as the target sequence, DNA extracted from the Salmonella strain was used as a template for fluorescence detection. Results: In this study, the detection of 7 related strains, in addition to Salmonella appeared very positive, the other strains were negative; in pure conditions, the quantitative detection limit of 30 cfu / ml; the same sample detected 3 times The coefficient of variation of Ct value was less than 5%. The coincidence rate of the two was 100% for the comparison between simulated and isolated culture. Conclusion: The establishment of this method not only provides a basis for the rapid detection of food-borne Salmonella contamination and food poisoning, but also directly for the detection of clinical stool specimens.