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1、JournalofVisualizedExperimentswww.jove.comVideoArticleTitrationofHumanCoronavirusesUsinganImmunoperoxidaseAssay1111FrancineLambert,HeleneJacomy,GabrielMarceau,PierreJ.Talbot1LaboratoryofNeuroimmunovirology,INRS-InstitutArmand-FrappierURL:https://www.jove.com/video/751DOI:doi:10.3791/751Keywords:M
2、icrobiology,Issue14,SpringerProtocols,Humancoronavirus,HCoV-229E,HCoV-OC43,cellandtissuesample,titration,immunoperoxidaseassay,TCID50DatePublished:4/28/2008Citation:Lambert,F.,Jacomy,H.,Marceau,G.,J.Talbot,P.TitrationofHumanCoronavirusesUsinganImmunoperoxidaseAssay.J.Vis.Exp.(14),e751,doi:10.3791
3、/751(2008).AbstractDeterminationofinfectiousviraltitersisabasicandessentialexperimentalapproachforvirologists.Classicalplaqueassayscannotbeusedforvirusesthatdonotcausesignificantcytopathiceffects,whichisthecaseforprototypestrains229EandOC43ofhumancoronavirus(HCoV).Therefore,analternativeindirecti
4、mmunoperoxidaseassay(IPA)wasdevelopedforthedetectionandtitrationofthesevirusesandisdescribedherein.Susceptiblecellsareinoculatedwithseriallogarithmicdilutionsofvirus-containingsamplesina96-wellplateformat.Afterviralgrowth,viraldetectionbyIPAyieldstheinfectiousvirustiter,expressedas'TissueCultureI
5、nfectiousDose50percent'(TCID50).Thisrepresentsthedilutionofavirus-containingsampleatwhichhalfofaseriesoflaboratorywellscontaininfectiousreplicatingvirus.ThistechniqueprovidesareliablemethodforthetitrationofHCoV-229EandHCoV-OC43inbiologicalsamplessuchascells,tissuesandfluids.Thisarticleisbasedonwo
6、rkfirstreportedinMethodsinMolecularBiology(2008)volume454,pages93-102.VideoLinkThevideocomponentofthisarticlecanbefoundathttps://www.jove.com/video/751/ProtocolThecompletetextprotocolforthisexperimentalapproachisavailableinSpringerProtocols.DisclosuresTheauthorshavenothingtodisclose.Copyright©200
7、8JournalofVisualizedExperimentsandSpringerProtocolsApril2008
8、14
9、e751
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