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1、VeterinaryMicrobiology,7(1982)295--306295ElsevierScientificPublishingCompany,Amsterdam--PrintedinTheNetherlandsENZYME-LINKEDIMMUNOSORBENTASSAYFORTHEDETECTIONOFTHECORONAVIRUS-LIKEAGENTANDITSANTIBODIESINPIGSWITHPORCINEEPIDEMICDIARRHEAP.CALLEBAUT,P.DEBOUCKandM.PENSAERTLaboratoryofV
2、irology,FacultyofVeterinaryMedicine,StateUniversityofGent,Casinoplein24,B-9000Gent(Belgium)(Accepted15February1982)ABSTRACTCallebaut,P.,Debouck,P.andPensaert,M.,1982.Enzyme-linkedimmunosorbentassayforthedetectionofthecoronavirus-[ikeagentanditsantibodiesinpigswithporcineepidemic
3、diarrhea.Vet.Microbiol.,7:295--306.Anenzyme-linkedimmunosorbentassay(ELISA)wasdevelopedforthedetectionofthecoronavirus-likeagentinfecesofpigsnaturallyaffectedwithporcineepidemicdiarrhea(PED)orexperimentallyinfectedwiththeCV777isolate.Theassaywasspecificandmoresensitivethanelectr
4、onmicroscopy.AnELISAblockingassayisdescribedforthedetec-tionandtitrationofantibodies.SpecificantibodyformationwasdemonstratedinpigsexperimentallyinfectedwithCV777andinswinenaturallyaffectedwithPED.INTRODUCTIONAcoronavirus-likeagent(CVLA)isassociatedwithdiarrheaaffectingswineofal
5、lages.Thistypeofdiarrheahasbeencalled"porcineepidemicdiarrhea"(PED)(Pensaert,1981).Basedonitselectronmicroscopicappearance,theagent(isolateCV777)hasbeenproposedasamemberoftheCoronaviridaefamily(PensaertandDebouck,1978).Todate,infectionsbythisagenthavebeendiagnosedbythedemonstrat
6、ionofthevirusinintestinalepitheliumusingimmunofluorescence(IF)ongutsectionsandinfecesusingelectronmicroscopy(EM)(Deboucketal.,1981b).AntibodiestoCV777havebeendemonstratedbyindirectIFstainingandbyimmunoelectronmicroscopy(Pensaertetal.,1981).Allthesemethodsarerathercumbersomeandti
7、meconsumingforlargescalediagnosticandserologicalpurposes.Theepizootiol-ogyofCVLAhas,therefore,notbeenexaminedindetail.TheELISAhasfoundwideapplicationinthedetectionofantibodiesandsmallamountsofantigenandseemstobeparticularlysuitableforwidespreadepizootiologicalstudies.Theassayhas
8、beenusedforthedetectionofbovinecoronavirusinfec
