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1、AdvancePublicationTheJournalofVeterinaryMedicalScienceAcceptedDate:1Nov2016J-STAGEAdvancePublishedDate:14Nov20161Virology2Fullpaper34Title:CharacterizationofcaninecoronavirusspreadamongdomesticdogsinVietnam56Authors:DungVanNguyen1,YukataTerada1,ShoheiMinami1,KenzoYonemitsu1,7NaoNagata1,Th
2、anhDinhHaLe2,RyuseiKuwata1,HiroshiShimoda1andKen8Maeda1*9101LaboratoryofVeterinaryMicrobiology,JointFacultyofVeterinaryMedicine,11YamaguchiUniversity,1677-1Yoshida,Yamaguchi753-8515,Japan122VeterinaryDiagnosticLaboratory,Sub-DepartmentofAnimalHealthofHoChiMinh13City,151LyThuongKiet,Distri
3、ct11,HoChiMinhCity,Vietnam1415*CorrespondingAuthor:KenMaeda,Ph.D.,D.V.M.16Tel&Fax:+81-83-933-588717E-mail:kmaeda@yamaguchi-u.ac.jp18RUNNINGHEAD:VIETNAMESECANINECORONAVIRUSES19120ABSTRACT.Caninecoronavirus(CCoV)isanimportantpathogenthatcauses21enteritisindogs,butthereisnoinformationonCCoVi
4、nfectioninVietnam.To22examinetheprevalenceofCCoVinfectionamongVietnamesedogs,201serum23sampleswereanalyzedbyvirus-neutralization(VN)test.Theresultsshowedthat24antibodyagainstCCoV-IIwaspresentin87dogs(43.3%).TodetectgenesofCCoV,25fecalsamplescollectedfrom30diarrheicand50healthydogswereexam
5、inatedbyRT-26PCR,confirmingthat2diarrheicdogsand5healthydogswerepositiveforCCoV.27NucleotidesequencesofN-terminalregionofspike(S)geneindicatedthatCCoV28strainsweredividedintotwosubgenotypes,CCoV-IIaand-IIb,respectively.29Furthemore,wesucceededinisolatingCCoV/dog/HCM47/2015,theisolatewas30
6、plaque-purifiedthreetimes,and3'-terminalone-thirdofthegenomewasanalyzed.31Interestingly,theplaque-purifiedvirushadalargedeletioninORF3abcandEgenes32(1,165nt),andashortdeletioninORF7bgene(60nt),suggestingthattheseregions33arenotnecessaryforinvitroreplicationofCCoV.Next,theantigenicitybetwe
7、enthe34isolatedCCoV-IIbandtheotherCCoV-IIawascomparedbyVNtest,revealingthat35antigenictyoftheisolatedCCoVisequalorhigherthanthatoftheotherCCoV.In36summary,twosubgenotypesofCCoV-IIarespreadingamongVietnamesedogs.The37isolatedviruswithalargedeletionafterinvitropassage
