三重PCR(聚合酶链式反应)检测鸡肉中常见病原菌的研究

Multiple PCR Rapid Detection for Three Pathogenic Bacteria in Chicken

建立鸡肉中常见3种致病菌即小肠耶尔森氏菌、大肠杆菌O157:H7和沙门氏菌的多重PCR(聚合酶链式反应)检测方法。方法:分别选择小肠耶尔森氏菌、沙门氏菌和大肠杆菌O157:H7基因组中高特异性的ail基因、inv A基因和ECs3032,设计并筛选具有高特异性的引物用于PCR(聚合酶链式反应)扩增。建立一种此3种致病菌的三重PCR检测方法,并通过人工污染实验对该法进行验证。结果:3对引物能特异地扩增出251、347、469 bp的目的条带;灵敏度:对这3种致病菌的单菌培养物检测灵敏度均为101CFU/m L。同时检测3种菌的灵敏度为103 CFU/m L。结论:该检测方法高效、精确、特异性强、灵敏度高,6 h^8 h内可同时快速检测多种致病菌,节省了大量时间。在实际检测中具有很好的实用价值和开发前景,可在食品检测和临床检验等领域大力推广。

A multiplex PCR method to detecte the Yersinia enterocolitica,E. coli O157： H7 and Salmonella in chicken meat was Established. The ail gene sequence of Yersinia enterocolitica,the ECs3032 gene sequence of E. coli O157： H7 and the inv A gene sequence of Salmonella were designed for primers and chose the high specific primers. A multiplex PCR method and demonstrated the method by artificial contamination experiments was Established. The primers amplified 251 bp, 347 bp, 469 bp target band specifically. The sensitivity of single bacterial detection was 101CFU/m L. The sensitivity of all bacterial was 103CFU/m L. The method was efficient, accurate, high specificity and sensitivity. The method which detect a variety of pathogens costs 6 h-8 h and saves a lot of time. The method had good practical value and development future in the actual testing. It should promote vigorously in the field of food testing and clinical examination.