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1、Jpn.J.Infect.Dis.,65,270-272,2012LaboratoryandEpidemiologyCommunicationsDetectionofHumanCoronavirusNL63andOC43inChildrenwithAcuteRespiratoryInfectionsinNiigata,Japan,between2010and2011MiyakoKon1,KaoriWatanabe1,TakashiTazawa1,KanakoWatanabe1,TsutomuTamura1,HiroyukiTsukagoshi2,MasahiroNoda3,HirokazuKi
2、mura3,andKatsumiMizuta4*1NiigataPrefecturalInstituteofPublicHealthandEnvironmentalSciences,Niigata950-2144;2GunmaPrefecturalInstituteofPublicHealthandEnvironmentalSciences,Gunma371-0052;3InfectiousDiseaseSurveillanceCenter,NationalInstituteofInfectiousDiseases,Tokyo208-0011;and4YamagataPrefecturalIn
3、stituteofPublicHealth,Yamagata990-0031,JapanCommunicatedbyMakotoTakeda(AcceptedMarch1,2012)Humancoronavirus(HCoV)isamemberoftherespiratoryviruses,includinginfluenzavirus,para-respiratoryvirusesthatincludesHCoV-229E,HCoV-influenzavirus,RSvirus,humanmetapneumovirus,OC43,andthesevereacuterespiratorysyn
4、dromerhinovirus,adenovirus,andenterovirus,from376(SARS)CoV(1).Recently,newtypesofHCoVs,suchspecimensusing6celllines(MDCK,LLC-MK2,CaCo-asNL63andHKU-1,havealsobeendescribed(1).2,HEp-2,Vero9013,andRD-18S).Therfore,weinves-Historically,HCoVresearchhasbeenhamperedbytigatedthepresenceofHCoVinthe131specime
5、nsfrompoorgrowthandlackofcytopathiceffectincellculturewhichnootherrespiratoryviruswasisolated.(1).ThedevelopmentofpolymerasechainreactionViralnucleicacidwasextractedfromthespecimens(PCR)technologyhadallowedthefieldofcoronavirol-usingtheQIAampViralRNAMiniKit(Qiagen,Ger-ogytodevelopwidelyandrapidly(1)
6、.However,accord-mantown,Md.,USA),suspendedinAVEbuffer,andingtotheInfectiousAgentsSurveillanceReport,onlyappliedtoRTreactionsusingaPrimeScriptTMRTRe-54and59HCoV-positivecases,including18casesfromagentKit(TakaraBio,Shiga,Japan)accordingtotheNiigata,werereportedinJapanin2010and2011,re-manufacturer'sins
7、tructions.Then,wescreenedforthespectively(2).Althoughwehavesucceededinisolatingamplificationof4HCoVs(HCoV-229E,HCoV-OC43,HCoV-229EvirusesfromchildrenwithnasopharyngitisNL63,andHKU-1)byperformingmultip
