简介：ThepurposeofthisstudywastotesttheeffectivenessofsourcevirusstrainforthemanufactureoftheinactivatedSARSvirusvaccine,andestablishanexperimentalmethodandpreliminarystandardforpotencyevaluation.MiceweredividedintogroupsforbeingimmunizedwithcorrespondingseriallydilutedexperimentalSARSvirusinactivatedvaccine.Andtherabbitswereimmunizedwithundilutedvaccine.ChallengeassaywasconductedwithaheterologousSARSvirus.Andtheneutralizationantibodywasdeterminedwithplaquereductionneutralizationtest(PRNT),towhichtheneutralizationantibodyintheconvalescentserumofSARSpatientswascompared.Theexperimentalvaccineviralstrainswereprovedtobesuitableformanufacturingthevaccine.Miceimmunizedbyvaccinesofserialdilutionswereabletoelicitneutralizingantibody.Theantibodytiterfrommiceimmunizedwiththeundilutedvaccinecouldreachupto1:495.2,whilethoseofrabbitsimmunizedwiththeundilutedvaccinecouldreachaGMTof55.0-79.9.ThecapabilityoftheantibodytoneutralizethevirusfromGuangdongismoreefficientthanthatfromBeijing.TheGMTofneutralizingantibodyinSARSconvalescentslivinginsouthandnorthChinarangedfrom50.12to54.95,andthetitersofconvalescentsfromnorthChinawerehigherthanthosefromsouthChina.Miceandrabbitsusedasthemodelforevaluationofpotencyareofsensitivity,andthetestisofreproducibility.Thecandidatechallengeviralstrainsshowedarelativelyconsistenteffectonevaluatingantibodiesproducedbyvariousbatchesanddifferentvaccine-sourcestrains,hencetheycanbeusedtoevaluatepotencyofthevaccine.Themethodfortestingthevaccinepotencyandtheevaluationstandardwasestablishedpreliminarily.

简介：Toestablishasensitiveandspecificmethodforseroepidermiologicaldetectionofhumanher-pesvirus8(HHV-8)infection,threepotentantigenicproteinsencodedbyopenreadingframes(ORFs)K8.1,65and73CingenomeofHHV-8wereproducedasglutathioneS-transferasefusionproteinintheprokaryoticexpressionsystemandwasusedasantigenfortesting.Therecombinantfusionproteinex-pressedintheprokaryoticexpressionvectorE.coliBL21waspurifiedbyglutathioneSepharose4Baffin-itychromatographyandwasquantitatedwithSDS-PAGE.Allthese3fusionproteinsproducedinthepro-karyoticexpressionsystemshowedgoodimmunogenicityasdemonstratedbyWesternblottingandcouldberecognizedbymixedseraofpatientswithKaposi′ssarcoma(KS).Theimmuno-reactivitiesofthesingleorcompoundfusionproteinweredeterminedbymeansofELISAandcomparedwiththetraditionalimmu-nofluorescenceassay(IFA)todeterminetheirsensitivityandspecificityofthetest.Itwasdemonstratedthatthesensitivityofmixed-antigenELISAmethodwassignificantlyhigherthanthatofIFA(81.8%vs34.4%),whilethespecificityoftheformerwasdemonstratedtobe97.9%.Thecoincidenceofthede-tectionratebetweenthesetwomethodswasconsiderablyhigh,approachingupto90.0%.TheseresultssuggestthatthemixedantigenELISAassayappearstobeasensitiveandspecificmethodforsero-epide-miologicaldetectionofhumanherpesvirus8infection.