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目的建立快速、准确检测食品中沙门菌、志贺菌、金黄色葡萄球菌和溶血性链球菌的多重荧光PCR方法。方法将多重荧光PCR技术与HANDS系统结合,采用同源加尾的方式,构建四重荧光PCR体系,分析该体系的特异性,并进行模拟实验,该方法与国家标准检测方法进行方法比对实验。结果该系统检测出了4种目标致病菌。初始菌浓度分别为5、9、10、2、50、3cfu/ml的沙门菌、志贺菌、金黄色葡萄球菌和兰氏A群、C群、G群链球菌增菌液,在增菌18h后都被体系检出。该方法与国家标准方法对样品的检测结果一致。结论首次结合HANDS系统构建了同时检测4种致病菌的多重荧光PCR方法,该方法特异性强、灵敏度高,是检测沙门菌、志贺菌、金黄色葡萄球菌和溶血性链球菌的快速、准确方法。
Objective To establish a rapid and accurate multi-fluorescence PCR method for the detection of Salmonella, Shigella, Staphylococcus aureus and hemolytic streptococcus in food. Methods The multiplex fluorescence PCR technique was combined with HANDS system. By using homologous tailed method, a four-fluorescence system was constructed. The specificity of the system was analyzed and a simulation experiment was carried out. The method was compared with the national standard detection methods . Results The system detected four kinds of target pathogens. Salmonellae, Shigella, Staphylococcus aureus and Rank A, C and G streptococci with initial bacterial concentrations of 5, 9, 10, 2, 50 and 3 cfu / ml, After 18h were detected by the system. The method and the national standard method for the detection of the same sample. Conclusions The multiplex fluorescent PCR method for simultaneous detection of four pathogenic bacteria was constructed for the first time in combination with HANDS system. The method is characterized by strong specificity and high sensitivity, and is a rapid, sensitive and sensitive method for the detection of Salmonella, Shigella, Staphylococcus aureus and Streptococcus hemolyticus. Accurate method