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1、多重PCR快速检测水产品中3种食源性致病菌方法的建立夏慧丽1基金项目：浙江省质监系统科研计划项目（20090237）*第一作者简介：夏慧丽（1978-），女（汉），高级工程师，博士，研究方向：食品质量检测与分析。联系电话：0576-88320885；E-mail：huili2001@163.com。郑辉台州市质量技术监督检测研究院浙江台州318000摘要：本文基于沙门氏菌(Salmonellatypli，St)侵袭性抗原保守基因invA、副溶血性弧菌(Vibrioparahaemolyticus,Vp)目的基因irgB和单核细胞增生李斯特菌(Listeriamonocyt
2、ogenes,Lm)调控蛋白基因prfA，设计了3对特异性引物，建立了快速检测水产品中3种主要食源性致病菌的多重PCR方法。结果显示，沙门氏菌、副溶血性弧菌和单核细胞增生李斯特菌的多重PCR扩增片段产物大小分别为213、369和517bp。反应条件优化后，3种目的菌在103CFU/mL均可同时扩增出较清晰条带，彼此之间无交叉反应，具有良好的特异性和灵敏度。该方法应用于3份人工污染水产样品的检测，并与国标方法对比验证，3种致病菌的整体符合率均达100%。该方法操作简单，检测周期短，灵敏度高、特异性强，能够实现对水产品中沙门氏菌、副溶血性弧菌和单核细胞增生李斯特菌3种食源性致
3、病菌的快速诊断检测和监控。关键词：多重PCR；沙门氏菌；副溶血性弧菌；单核细胞增生李斯特菌；水产品Establishmentofmultiple-PCRfordiagnosingthreefoodbornepathogenicbacteriainaquaticproductsXiaHui-liZhengHuiResearchInstituteofQuality&TechnicalSupervisionandInspection,Taizhou,Zhejiang,318000,ChinaAbstract:Arapidmultiplex-PCRmethodwasestabli
4、shedinordertodetectthreefoodbornepathogenicbacteriaincludingSalmonellatypli，VibrioparahaemolyticusandListeriamonocytogenesinaquaticproducts.Threepairsofoligonucleotideprimersweredesignedformultiplex-PCRamplificationaccordingtogenecodinginvasionproteinAofSalmonellatypli，genecodingimmuneres
5、ponseofVibrioparahaemolyticusandregulationhistonegeneofListeriamonocytogenes.geneofShigellaspp．TheamplifiedfragmentsizesofthesethreebacteriawerelocatedintheTomb,DongShenJiabang,deferthenextdayfocusedontheassassination.Linping,Zhejiang,1ofwhichliquorwinemasters(WuzhensaidinformationisCarpe
6、nter),whogotAfewbayonets,duetomissedfatal,whennightcame7213bp,369bpand517bp,respectively．Undertheoptimizedconditions,thespecificityofthemultiplex-PCRwashigh,andtheminimumdetectionlimitwas103CFU/mL.Themultiplex-PCRmethodwasusedtoanalyzethreeaquaticproductsamplescomparedwithnationalstandard
7、methods,thecoincidencerateoftwomethodsreached100%.Themethoddevelopedinthisstudyhadhighsensitivityandspecificity,whichcouldbeappliedfortherapiddetectionandmolecularepidemiologysurveyoffoodbornepathogenicbacteriainaquaticproducts．Keywords:Mmultiple-PCR,Salmonellatypli
