【摘要】 建立了同步快速检测食品中志贺氏菌、沙门氏菌、变形杆菌的多重聚合酶链式反应(multiplex polymerase chain reaction,mPCR)的方法。通过已报道基因和序列比对确定3种致病菌的特异性基因-志贺氏菌的ipaH基因、沙门氏菌的invA基因和变形杆菌的atpD基因并设计引物,优化反应条件,建立3种致病菌的多重PCR检测体系,测定方法特异性和灵敏度。结果表明,建立的多重PCR方法灵敏度为102CFU/mL,人工模拟样品的灵敏度为103CFU/mL。初步建立了能同步、简便、快速、灵敏地检测食品中志贺氏菌、沙门氏菌、变形杆菌的三重PCR方法,可为卫生检验检疫、食品中致病菌的监测和监督提供较为实用的方法。更多还原

【Abstract】 To develop a rapid multiplex polymerase chain reaction(m-PCR) assay for simultaneous detection of Shigella spp.,Salmonella spp.and Proteus vulgaris in food.The genomic alignment method was used to identify specific PCR target sequences.Design the three pairs of specific primers.IpaH gene of Shigella spp.,invasive protein gene(invA) of Salmonella spp.,and atpD gene of Proteus vulgaris Multiplex PCR was established by optimizing the reaction system.The specificity and sensitivity of the new multiplex PCR system were also evaluated.The result showed that the sensitivity of the multiplex PCR method was 102 CFU/mL,and the sensitivity of artificial pollution was 103 CFU/mL.The conclusions indicated that a triple PCR assay has been established for the simultaneous,sample,rapid and sensitive detection of Shigella spp.,Salmonella spp.and Proteus vulgaris in food which also offers a convenient and rapid alternate microbiological tool for the detection and surveillance of sanitary inspection quarantine and food microbiological safety.更多还原