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1、veterinarymicrobiologyELSEVIERVeterinaryMicrobiology42(1994)349-359Detectionofporcinerespiratorycoronavirusandtransmissiblegastroenteritisvirusbyanenzyme-linkedimmunosorbentassayE.Cornaglia*,N.Chr6tien,S.Charara,Y.ElazharyVirologySection.FacultyofVeterinaryMedicine,Uni
2、versityofMontreal,P.O.Box5000,SaintHyacinthe,Que..J2S7C6,CanadaReceived24November1993;accepted17May1994AbstractAnenzyme-linkedimmunosorbentassay(ELISA)forthedetectionoftransmissiblegastroenteritisvirus(TGEV)andporcinerespiratorycoronavirus(PRCV)wasdeveloped.AbovineTGEV
3、-specificpolyclonalantibodywaspurifiedbyaffinitychromatographywiththeTRIOBioprocessingSystemandwasusedasthecaptureantibody,ataconcentrationof1.5txg/well.TheF5.39monoclonalantibodywasobtainedbythefusionofspleenlymphocytesfromTGEVimmunizedmicewithNS-Imyelomacells.ThismAb
4、wasusedasasecondantibodyfortheELISA.TheELISAdetected40ngofTGEVand407ngofPRCV.TostudytheabilityofELISAtodetectTGEVinfieldcases,53intestinalsamplesweretakenfrompigsexhibitingclinicalsignsoftransmissiblegastroenteritis.AllthepositivesamplesdetectedbytheELISAwereconfirmeda
5、spositivebyimmunofluorescenceorcellcultureimmunofluorescence.TostudytheabilityofthisELISAtodetectPRCVinnasalswabsandlungsamples,20seven-day-oldpigletswereinoculatedwithaQuebecstrainofPRCV.TheELISAwasabletodetectPRCVinbothkindsofsamples.Keywords:Porcinerespiratorycorona
6、virus;Pig;Porcinetransmissiblegastroenteritisvirus;Diagnosis,virus;ELISA1.IntroductionTransmissiblegastroenteritis(TGE)wasfirstreportedintheU.S.in1945(DoyleandHutchings,1946)andsincethenithasbeenshowntohaveanalmostworldwidedistribution(SaifandHeckert,1990).TGEischaract
7、erizedbyvomiting,aprofusewaterydiarrhea,andahighmortalityrate(often100%)inpigletsunder2weeksofage,andmanifestsitself*Correspondingauthor.0378-1135/94/$07.00©1994ElsevierScienceB.V.AllrightsreservedSSD10378-1135(94)00076-9350E.Cornagliaetal./VeterinaryMicrobiology42(199
8、4)349-359astransitoryenteritisinadultpigs(SaifandHeckert,1990;Moxleyetal.,1993).Someserologicalsurveysrevealedthatsam
