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1、CELLCULTUREPROPAGATIONOFBOVINECORONAVIRUSLindaJ.Sail,RobertA.Heckert,KathyL.Miller,andMohamedM.TarekFoodAnimalHealthResearchProgram,OhioAgriculturalResearchandDevelopmentCenter,OhioStateUniversity,Wooster,Ohio44691,Ohio:andDepartmentofParasitologyandMicrob
2、iology(M.M.T.),FacultyofMedicine,ZagazigUniversity,Zagazig,EgyptSUMMARY:Althoughmostfieldstrainsofbovinecoronavirus(BCV)growpoorlyincellcultureandfailtoproducecytopathiceffects{CPE)untilafterblindpassage,primarycalfkidney{PCK)andVerocellshavepermittedprima
3、ryisolationofvirus.Cellculture-adaptedstrainsofBCVreplicateinPCK,bovineembryoniclung,bovinefetalthyroid,bovinefetalbrain,bovineskincells,ovinefetalkidneycells,andthecelllinespigkidneyK3and15,Vero,humanembryoniclungfibroblasts,HRT-18,MDBKandBEK-1,withtrypsi
4、nusefulforenhancingreplication.Organcultureaswellassucklingmouse,rat,andhamsterbrainsalsosupportthegrowthofcellculture-adaptedBCVstrains.ViralgrowthismostcommonlydetectedbyCPE,im-munofluorescence,hemagglutination,andhemadsorptionassaysorelectronmicroscopyo
5、fsupernatantsfrominfectedcells.Inthisreport,theoptimalconditionsforthegrowthandplaqueassayoftheNCDVstrainofBCVinMDBKcellsaredescribed.Keywords:bovinecoronavirus;cellculture;propagation.I.INTRODUCTIONInthisreport,theoptimalcellcultureconditionsforthegrowtha
6、ndplaqueassayoftheNCDV{Mebus)Thecoronaviruseswerefirstrecognizedandmor-strainofBCVinMDBKcellsaredescribed.Alsoin-phologicallydefinedasagroupbyTyrrelletal.,(39-41jcludedisareviewoftheliteraturepertainingtothebuthavehavebeenknownbyothernamesforalmost5cultiva
7、tionofBCVsincellandorgancultures.decades(33).TheCoronaviridaeisamonogenericfamilycomprising11virusesthatinfectvertebrates(29).II.MATERIALSStairetal.(30)firstdescribedtheisolationandpartialcharacterizationofaeoronavirus-likeagentfromcalvesA.Equipmentwithdia
8、rrhea.Theviralparticleswere107to160nm,VortexMixer,modelK-550-G,ScientificIndustries1polymorphic,andhadanll-nmpetal-shapedfringe.Incubator,32fP,NationalAppliance2Sharpeeetal.{28)furthercharacterizedthisIncubat
