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六种牛源致病性细菌多重PCR检测方法的建立.docx

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六种牛源致病性细菌多重PCR检测方法的建立 摘要： 本研究建立了一种能够同时检测六种牛源致病性细菌的多重PCR检测方法，包括沙门氏菌、大肠杆菌、肺炎克雷伯菌、铜绿假单胞菌、葡萄球菌和链球菌。使用该方法检测了牛粪和奶样品中的细菌，结果表明该方法具有高灵敏度和特异性。本研究的多重PCR检测方法可在养殖业和食品安全领域中广泛应用。 Abstract: Inthisstudy,amultiplexPCRdetectionmethodwasestablishedforthesimultaneousdetectionofsixbacterialpathogensfromcattlesources,includingSalmonella,Escherichiacoli,Klebsiellapneumoniae,Pseudomonasaeruginosa,Staphylococcusaureus,andStreptococcusagalactiae.Bacteriainbovinefecalandmilksamplesweretestedusingthismethod,andtheresultsshowedthatthemethodhadhighsensitivityandspecificity.ThemultiplexPCRdetectionmethoddevelopedinthisstudycanbewidelyusedinthefieldofanimalhusbandryandfoodsafety. Introduction: Livestockfarmingisanimportantindustryinmanycountriesworldwide.Theproductionofbeef,milk,andotheranimal-derivedproductsisamajorcontributortotheeconomy.However,bacterialinfectionscanresultinsignificanteconomiclossesforthelivestockindustryaswellasposeapublichealthrisk.ThebacteriamostcommonlyassociatedwithinfectionsincattleincludeSalmonella,Escherichiacoli,Klebsiellapneumoniae,Pseudomonasaeruginosa,Staphylococcusaureus,andStreptococcusagalactiae.Therefore,developinganefficientandreliablemethodfordetectingthesebacterialpathogensisofgreatimportanceforthelivestockindustry. Polymerasechainreaction(PCR)isawidelyusedmethodforthedetectionofbacterialpathogensduetoitshighsensitivityandspecificity.However,conventionalPCRassayscanonlydetectonetargetatatime.MultiplexPCR,atechniquethatcandetectmultipletargetsinasinglereaction,hasbeenextensivelyusedtoovercomethislimitation.Bycombiningmultipleprimerpairsthatarespecifictoeachtarget,itispossibletodetectthepresenceofseveralbacterialpathogensinasinglereaction. Inthisstudy,weaimedtoestablishamultiplexPCRmethodtosimultaneouslydetectsixbacterialpathogensfromcattlesources:Salmonella,Escherichiacoli,Klebsiellapneumoniae,Pseudomonasaeruginosa,Staphylococcusaureus,andStreptococcusagalactiae. MaterialsandMethods: Bacterialstrainsandgrowthconditions: Bacterialstrainsusedinthisstudywerepurcha