预览加载中，请您耐心等待几秒...

基于非洲猪瘟病毒pA104R蛋白抗体检测间接ELISA方法的建立.docx

预览
1/2
2/2

在线预览结束，喜欢就下载吧，查找使用更方便

下载提示 文本预览

如果您无法下载资料，请参考说明：

1、部分资料下载需要金币，请确保您的账户上有足够的金币

2、已购买过的文档，再次下载不重复扣费

3、资料包下载后请先用软件解压，在使用对应软件打开

基于非洲猪瘟病毒pA104R蛋白抗体检测间接ELISA方法的建立 Title:EstablishmentofIndirectELISAMethodforDetectionofAntibodiesagainstAfricanSwineFeverVirusProteinpA104R Abstract: Africanswinefever(ASF)isahighlycontagiousviraldiseaseaffectingdomesticandwildpigs,resultinginsevereeconomiclossesworldwide.Effectiveandaccuratedetectionmethodsarecrucialforearlydiagnosisanddiseasecontrol.Thisstudyaimstoestablishanindirectenzyme-linkedimmunosorbentassay(ELISA)methodforthedetectionofantibodiesagainstthepA104RproteinoftheAfricanswinefevervirus(ASFV). Introduction: Africanswinefever(ASF)iscausedbytheAfricanswinefevervirus(ASFV),alargedouble-strandedDNAvirus.ASFposesasignificantthreattoglobalporkproductionandfoodsecurityduetoitshighmortalityrateandlackofaneffectivevaccine.ThepA104RproteinisoneofthemajorantigenicproteinsofASFVandhasbeendemonstratedasapotentialtargetforserologicaldetection. Methods: 1.ProductionandpurificationofpA104Rprotein.ThepA104Rproteinwasexpressedinasuitableexpressionsystemandpurifiedusingaffinitychromatography. 2.Preparationofcoatingantigen.ThepurifiedpA104RproteinwasimmobilizedonELISAplatesasthecoatingantigen. 3.Serumsamplepreparation.SerumsamplesfromASFV-infectedpigsanduninfectedcontrolpigswerecollectedandprocessedforantibodydetection. 4.OptimizationofindirectELISAconditions.Keyfactorssuchasserumdilutions,antigenconcentration,blockingbuffer,andincubationtimewereoptimizedtoestablishanoptimalindirectELISAprotocol. 5.EvaluationoftheindirectELISAmethod.Sensitivity,specificity,andcross-reactivityoftheestablishedmethodwereevaluatedusingknownpositiveandnegativeserumsamples.Comparisonwithotherdetectionmethods,suchasvirusneutralizationtest(VNT)andpolymerasechainreaction(PCR),wasalsoconducted. Results: ThepA104Rproteinwassuccessfullyexpressedandpurified.TheoptimizedindirectELISAprotocolexhibitedhighsensitivityandspecificityfordetectingantibodiesagainstASFVinpigserumsamples.Thepositiveandnegativecontrolsshowedexpectedresults,indicatingthereliabilityoftheestablishedmethod.ComparisonwithVNTandPCR