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1、J.gen.ViroL(I98o),49,83-8983PrintedinGreatBritainEnzyme-linkedImmunosorbentAssayforCoronavirusesHCV229EandMHV3ByCORNELISA.KRAAIJEVELD,M.HILARYMADGEANDMALCOLMR.MACNAUGHTONDivisionofCommunicableDiseases,ClinicalResearchCentre,Harrow,MiddlesexHA13UJ,U.K.(AcceptedIFebruaryI98O)SUMMARYTheantigenicrelat

2、ionshipbetweenhumancoronavirusstrainz29E(HCVza9E)andmousehepatitisvirusstrain3(MHV3)wasstudiedbymeansoftheindirectformofenzyme-linkedimmunosorbentassay(ELISA).Across-reactionwasfoundwithhyperimmunerabbitserabetweenHCV229EandMHV3whichmaybeduetotheadherenceofbovineserumcomponentsfromtissueculturemed

3、ia,whichwerepresentonvirusparticlesevenafterextensivepurification.Nocross-reactionwasobservedwithimmuneseraabsorbedwithbovineserum,orwithHCV229Egrownintissueculturewithoutserum.ThisindirectELISAwithHCV229Emayprovetobeusefulforstudieswithhumansera.INTRODUCTIONTheenzyme-linkedimmunosorbentassay(ELIS

4、A)wasfirstdescribedbyEngvall&Perlmann(I97I)andvanWeemen&Schuurs(I970.Sincethenvariationsofthetesthavefoundwideapplicationinthedetectionofantibodiesandsmallamountsofantigen(Bidwelletal.I977).Indiagnosticserology,enzyme-immunoassaysarehighlysensitiveandrelativelyeasytoperformandseemtobeespeciallysui

5、tedtolarge-scaleepidemiologicalsurveys.However,thehighsensitivityofELISAmeansthatthespecificityoftheantigensusedisofmajorimportance.Detectionofantibodiesagainstcoronavirusesbyneutralization,complementfixation,immunofluorescenceorimmunodiffusioniscumbersomeandgenerallynotveryreliable(Mclntosh,I974)

6、.Therefore,weattemptedtoseeifanELISAtechniquewithhumancoronavirusstrain299E(HCV299E)andmousehepatitisvirusstrain3(MHV3)couldbeusedasasensitive,specifictestfordetectingantibodiesinanimalhyperimmunesera.Bradburne(197o)foundacross-reactionbetweenHCV229EandMHV3withhyper-immuneanimalseraincomplementfix

7、ationandimmunodiffusiontests.Cross-reactionsbetweentheseviruseswerenotfoundbyotherworkers(Mclntoshetal.1969;Pedersenetal.~978).AntibodiesreactingwithMHVhavebeendetectedinhumanseraandareprobablyresponsestorelatedh