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1、ArchVirol(2001)146:2401–2419Cross-protectionstudiesbetweenrespiratoryandcalfdiarrheaandwinterdysenterycoronavirusstrainsincalvesandRT-PCRandnestedPCRfortheirdetectionK.-O.Cho∗,M.Hasoksuz∗∗,P.R.Nielsen,K.-O.Chang,S.Lathrop∗∗∗,andL.J.SaifFoodAnimalHealthResearchProgram,DepartmentofVeterinaryPreventi
2、veMedicine,OhioAgriculturalResearchandDevelopmentCenter,TheOhioStateUniversity,Wooster,Ohio,U.S.A.AcceptedJanuary17,2001Summary.A1-stepRT-PCRassay,targetinga730bpfragmentofthenucleo-capsid(N)geneofbovinecoronavirus(BCV),andanestedPCRassay,targetinga407bpfragmentoftheNgene,weredevelopedtodetectBCVi
3、nnasalswabandfecalsamplesofcalvesexperimentallyexposedtoBCV.Both1-stepRT-PCRandnestedPCRrecognizedcellculturepassagedisolatesof10bovinerespiratorycoronavirus(BRCV),5calfdiarrhea(CD)and8winterdysentery(WD)strainsofBCV,butnottransmissiblegastroenteritiscoronavirusorbovinerotavirus.Thesensitivityofth
4、e1-stepRT-PCRandnestedPCRwascomparedtothatofanantigen-captureELISA.Thelowestdetectionlimitofthe1-stepRT-PCRandnestedPCRasdeterminedbyusingtenfoldserialdilutionsoftheBRCV255and440strainsinBCVnegativenasalswabsuspensionsfrompreexposuregnotobi-oticcalveswas2×104and2×102TCID/0.1mlforeachstrain,respect
5、ively.50Thelowestdetectionlimitoftheantigen-captureELISAasdeterminedbyusingthesameseriallydilutedsampleswas1×106TCID/0.1mlforeachstrain.50Therefore,the1-stepRT-PCRandnestedPCRassayswere50and5000times,respectivelymoresensitivethantheantigen-captureELISAtodetectBRCVinnasalswabsuspensions.Toinvestiga
6、teinvivocross-protectionbetweentheBRCVandCDorWDstrainsofBCVandtodetectnasalandfecalshedding∗Presentaddress:CollegeofVeterinaryMedicine,ChonnamNationalUniversity,Kwangju500-757,SouthKorea.∗∗Permanentaddress:IstanbulUniversity,VeterinaryFaculty,DepartmentofMicro-biology,Avcilar,34851,Istanbul,Turkey
7、.∗∗∗Presentaddress:EISO,BacterialZoonosesBranch,CentersofDiseaseControlandPrevention’sNationalCenterforInfectiousDiseases,Division,FortCollins,CO,U.S.A.2402K.-O.Choetal.ofBCVusingthe1-stepRT-PCR,nestedPCRandantig
