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1、JVetDiagnInvest11:15±19(1999)SensitivitycomparisonfordetectionofrespiratorybovinecoronavirusesinnasalsamplesfromfeedlotcattlebyELISAandisolationwiththeGcloneofHRT-18cellsManuelReisdaSilva,KathyL.O'Reilly,XiaoqingLin,LisaStine,JohannesStorzAbstract.Amonoclonalant
2、ibody-basedcaptureenzyme-linkedimmunosorbentassay(ELISA)wasdevel-opedtodetectrespiratorybovinecoronavirus(RBCV)antigensinnasalswabscollectedfromcattleshowingsignsofrespiratorytractdiseasefollowingshipping.ThesesampleshadbeenpreviouslytestedforRBCVbyinoculationof
3、Gcloneculturesofhumanrectaltumorcells(HRT-18G)andforbovineherpesvirus1,para-in¯uenzavirus3,bovineadenovirus,bovinerespiratorysyncytialvirus,andbovineviraldiarrheavirusonotherspeci®callypermissivecellcultures.RBCVhasnotpreviouslybeenrecognizedasanimportantetiolog
4、icalfactorinthebovinerespiratorydiseasecomplexoffeedlotcattle.Thirtyof100samplestestedpositiveforRBCVantigenbycaptureELISAincontrastto38of100samplesthatyieldedRBCVisolatesinGclonecells.SamplesyieldingotherbovinerespiratoryvirusesintheabsenceofRBCVwerenegativeint
5、hecaptureELISA,whichwasbasedontheuseofasinglemonoclonalantibodythatrecognizesoneRBCVepitopeontheSglycoproteinwiththebroadestreactivitywithdifferentstrainsofRBCVtested.SomeRBCVstrainsmaynotbedetectedbythisELISA,whichmayaccountforthehigherpercentageofRBCV-infected
6、cattledetectedbyRBCVisolation.However,theELISAwassimpletoperform,sensitive,andspeci®candwasmorerapidthanvirusisolation.Thisassaywillbeusefulforprocessinglargenumbersof®eldsamplesinfutureepidemiologicanddiagnosticstudiesofRBCVinfectionsofcattle.Bovinerespiratoryd
7、iseaseisthesinglemostim-RBCVshavenotbeenisolatedpreviouslyfromfeedlotportantsyndromeaffecting6±8-montholdbeefcattleorotheradultcattlebyemployingconventionalcellafterentryintofeedlotsinNorthAmerica.7Bacteria,culturetechniques,yetthesevirusesmaybeacontrib-mycoplas
8、ma,andvirusessuchasbovineherpesvirusutingfactorinrespiratorytractdiseaseincattle.The1,parain¯uenzavirus3(PI-3),bovinerespiratorysyn-ELISAcanfacilitateprocessingoflarg
