冠状病毒Coronavirusl论文-1999 Evaluation of the Baculovirus-Expressed S Glycoprotein of Transmissible Gastroenteritis Virus (TGEV) as Antigen in.pdf

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1、JVetDiagnInvest11:205±214(1999)Evaluationofthebaculovirus-expressedSglycoproteinoftransmissiblegastroenteritisvirus(TGEV)asantigeninacompetitionELISAtodifferentiateporcinerespiratorycoronavirusfromTGEVantibodiesinpigsKarolSestak,ZhongfangZhou,DavidI.Shoup,Lind

2、aJ.SaifAbstract.Thespike(S)glycoproteinoftheMillerstrainoftransmissiblegastroenteritisvirus(TGEV)wasrecentlyclonedandexpressedinbaculovirus.TherecombinantSproteinwasusedasthecoatingantigeninacompetition(blocking)enzyme-linkedimmunosorbentassay(ELISA)incombinat

3、ionwithmonoclonalanti-bodiestotheSproteinepitopeA(conservedonTGEVandporcinerespiratorycoronavirus[PRCV])orepitopeD(presentonTGEVonly)todifferentiatePRCV-fromTGEV-inducedantibodies.Oneset(setA)of125serumsampleswerecollectedatdifferenttimesafterinoculationofcaes

4、arean-derived,colostrum-deprived(n552)andconventionalyoungpigs(n573)with1ofthe2porcinecoronavirusesoruninoculatednegativecontrols(TGEV/PRCV/negative575/30/20).Asecondset(setB)of63serumsamplesoriginatedfromadultsowsinoculatedwithPRCVandtherecombinantTGEVSprotei

5、norwithmock-proteincontrolandthenexposedtovirulentTGEVafterchallengeoftheirlitters.SerafromsetAwereusedtoassesstheaccuracyindicators(sensitivity,speci®city,accuracy)ofthe®xed-cellblockingELISA,whichusesswinetesticularcellsinfectedwiththeM6strainofTGEVastheanti

6、gensource(ELISA1)andthenewlydevelopedELISAbasedontherecombinantSproteinasantigen(ELISA2).TheserafromsetB(adults)weretestedforcomparison.Theplaquereductionvirusneutralizationtestwasusedasacon®rmatorytestforthepresenceofantibodiestoTGEV/PRCVinthetestsera.Theaccu

7、racyindicatorsforbothELISAssuggestthatdifferentialdiagnosiscanbeofpracticaluseatleast3weeksafterinoculationbytestingthedual(acute/convalescent)samplesfromeachindividualinconjunctionwithanothercon®rmatory(virusneutralization)antibodyassaytoprovidevalidandcomple

8、tedifferentiationinformation.Moreover,whereasELISA1had10±20%falsepositiveresultstoepitopeDforPRCV-infectedpigs(setAsamples),nofalse-positiveresultstoepitopeDoccurredusingELISA2,ind