冠状病毒Coronavirusl论文-1990 Enzyme-linked immunosorbent assay for the detection of porcine epidemic diarrhea coronavirus antibodies in swine se.pdf

冠状病毒Coronavirusl论文-1990 Enzyme-linked immunosorbent assay for the detection of porcine epidemic diarrhea coronavirus antibodies in swine se.pdf

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1、VeterinaryMicrobiology,21(1990)263-273263ElsevierSciencePublishersB.V.,Amsterdam--PrintedinTheNetherlandsEnzyme-linkedImmunosorbentAssayfortheDetectionofPorcineEpidemicDiarrheaCoronavirusAntibodiesinSwineSeraMARTINHOFMANNandROBERTWYLER*InstituteofVirology,U

2、niversityofZurich,Winterthurerstr.266a,CH-8057Zurich(Switzerland)(Acceptedforpublication21July1989)ABSTRACTHofmann,M.andWyler,R.,1990.Enzyme-linkedimmunosorbentassayforthedetectionofporcineepidemicdiarrheacoronavirusantibodiesinswinesera.Vet.Microbiol.,21:2

3、63-273.Anenzyme-linkedimmunosorbentassay(ELISA)fordetectingserumantibodiestothepor-cineepidemicdiarrheacoronavirus(PEDV)wasestablishedbyusingcellculture-grownPEDVasantigenforcoating.Ultracentrifugationthrough20and45%(w/w)sucrosecushionsprovedtobethebestanti

4、genpurificationmethod.Examinationof1024swineserashowedahighspeci-ficityandagreatersensitivityoftheELISA,whencomparedwithindirectimmunofluorescence.ReferenceserawithhighantibodytiterstoPEDVoriginatedfromtwopigsexperimentallyin-fectedwithPEDV.Threedifferentan

5、tigenpurificationmethodsandtheadvantagesoftheELISAcomparedwithanimmunofluorescencetestarediscussed.INTRODUCTIONPorcineepidemicdiarrheavirus(PEDV)(PensaertandDebouck,1978;De-bouckandPensaert,1980;Deboucketal.,1981;Witteetal.,1981)isidentifiedasthecausativeag

6、entleadingtodiarrheathroughdestructionoftheepithelialcellsofthesmallintestine.AlthoughithasbeenpossibletodemonstratePEDVinfectionsinmanycountries,extensiveserologicalstudieshavebeenhinderedbythelackofasimple,rapidandsensitivetesttodetectPEDVantibodiesinporc

7、inesera.ForPEDVserology,threetestshavebeendescribedintheliter-ature,namelyanindirectimmunofluorescencetest(IIFT)(PragerandWitte,1981a,b;HofmannandWyler,1987),animmunofluorescenceblockingtest(IFBT)(WitteandPrager,1987)andanenzyme-linkedimmunosorbentas-say(EL

8、ISA)blockingtest(Callebautetal.,1982).Allthesetestsrequirean-*Authortowhomallcorrespondenceshouldbeaddressed.0378-1135/90/$03.50©1990ElsevierSciencePublishersB.V.264M.HOFMANNANDR.WYLERtigenfromPEDV-inf

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