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1、JournalofMedicalVirology31:165-172(1990)CharacterizationofaNucleicAcidProbefortheDiagnosisofHumanCoronavirus2293InfectionsS.Myint,D.Harmsen,T.Raabe,andS.G.SiddellMRCCommonColdUnit,SalisburySP28BW,England(S.M.);fnstitutfurVirologieandImmunbiologiederUniuers
2、itatWurzburg,0-8700Wurzburg,FederalRepublicofGermany(D.H.,T.R.,S.G.S.)AcDNAcopyoftheHCV229Enucleocapsidpro-ThereplicationofHCV229Einvolvesthesynthesisteingenewasisolatedandcharacterized.se-ofa3'co-terminalsetofsixsubgenomicRNAs[Weissquenceanalysispredictsa
3、nucleocapsidpoly-andLeibowitz,19811.ItisassumedthattheseRNAspeptideof389aminoacidswithamoleculararesynthesizedinthecytoplasmofinfectedcellsbyaweight(mol.wt.)of43,450.SinglestrandRNAprocessofleader-primeddiscontinuoustranscription,probesderivedfromthecDNAco
4、pyhybridizeashasbeendescribedforthemurinehepatitisvirus,specificallytoHCV229ERNAandapproximatelyMHV[Baricetal.,1985;Makinoetal.,1986a;Shiehet50pgofintracellularviralRNAcanbereadilyal.,19871.Recently,theHCV229Egenesencodingthedetected.Theapplicationofnuclei
5、cacidhybrid-nucleocapsid,membrane,andspikeproteinshavebeenizationasaroutineprocedureforthediagnosisclonedandsequencedandtheirorderonthegenomeofHCV229Einfectionisdiscussed.hasbeendeterminedas5'S-M-N3'[RaabeandSid-dell,1989a;Raabeetal.,1990;Schreiberetal.,19
6、891.KEYWORDS:Ngenesequence,hybridizationRaabeetal.[1990]haveproposedthatthesubgenomicanalysis,coronavirusRNAs2,6,7functionasthemRNAsfortheS,M,andNproteins,respectively.Basically,threeapproachestothediagnosisofHCVINTRODUCTIONinfectionshavebeenused.Firstly,t
7、hereistheisolationandidentificationofHCVsincellandorganculture.Humancoronaviruses(HCVs)arecausativeagentsHowever,thisapproachistimeconsumingandre-ofrespiratoryillnessinman.Inhealthyadultstheyarequiresconsiderableexpertise.OnlyHCV229E-likevi-associatedwithc
8、ommoncoldsofmildtomoderatese-rusescanbedirectlyisolatedincellcultureandotherverity,usuallyof6-8daysduration.Thetypicalsymp-HCVshavetobeisolatedinorgancultureand,ifpos-tomsarenasalcatarrhandsorethroat,although