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1、ArchVirol(1999)144:167–175ExperimentalinoculationofadultdairycowswithbovinecoronavirusanddetectionofcoronavirusinfecesbyRT-PCRBriefReportH.Tsunemitsu1;2,D.R.Smith1;,andL.J.Saif11FoodAnimalHealthResearchProgram,OhioAgriculturalResearchandDevelopmentCenter,TheOhioStateUniversity,Wooster,Ohi
2、o,U.S.A.2HokkaidoResearchStation,NationalInstituteofAnimalHealth,Sapporo,Hokkaido,JapanAcceptedAugust5,1998Summary.AreversetranscriptasePCR(RT-PCR)targetinga407bpfragmentofthenucleocapsidgeneofbovinecoronavirus(BCV)wasdevelopedfordetec-tionofBCVRNAinfecesofexperimentallyinoculatedcattle.Th
3、esensitivityandspeci?cityoftheRT-PCRwerecon?rmedusingtissueculture-adaptedBCVstrainsandfecesof2calvesinoculatedwithBCV.Tennonpregant,BCVseropos-itive,adultdairycowswereinoculatedwithwinterdysentery(WD)(nD8)orcalfdiarrhea(CD)(nD2)strainsofBCVintranasallyandorally(nD2)orthroughasurgically-pl
4、acedduodenalcatheter(nD8)withandwithoutdexamethasonetreatmentorfeedingicewater.The6cowsinoculatedwithBCVintranasallyandthroughaduodenalcatheter(2of2cowsgivenCDBCVand4of6cowsgivenWDBCV)developedmilddiarrhea,andBCVwasdetectedindiarrhealfecesbyRT-PCR,ELISAorimmuneelectronmicroscopy.Theseresul
5、tssuggestthatCDandWDstrainsofBCVcancausediarrheainadultcowsinconjunctionwithhostorenvironmentalfactorsandthatRT-PCRmightbeusefultodiagnoseBCVinfectionsincalvesandadultcows.Bovinecoronavirus(BCV)isanimportantpathogenofneonatalcalfdiarrhea(CD)[5].Thevirusalsohasbeenisolatedfromadultcattleex
6、periencingacutedi-arrheaduringoutbreaksofwinterdysentery(WD)[19].Inarecentstudyindivid-Presentaddress:DepartmentofVeterinaryandBiomedicalSciences,UniversityofNebraska,Lincoln,NE68583,U.S.A.168H.Tsunemitsuetal.ualsinherdsexperiencingWDweremorelikelytohaverecentlyserorespondedtoBCVthanindiv
7、idualsinnon-affectedherds[22].However,theroleofBCVintheWDsyndromeremainsunclear.AfewattemptsweremadetoexperimentallyreproduceWDincowsusingfecesfromcowswithWDaschallengeinocula,butresultshavebeeninconsistent[3,28].CertainriskfactorscombinedwithBCVinfectionmight
