采用基因芯片检出一例丙型肝炎病毒3b基因亚型的报告

3b1,2,1,2,1,13(11,430071;21,430080):R373121:A:1000-8721(2003)03-0274-04:2002-10-31;:2003-05-20:(1977-),,,,(1954-),,,,,,E-mail:x1zhang@pentium1whiov1ac1cnTel/Fax:[phone],HCVHCV,HCVHCVChan5NCR,HCV123[1],CNS3NS5,,1a1b2a[2,3]Simmonds[4]HCV4,Chan[5]Houghton[6]1a1b;Enomoto[7]2a2b;Cha[8]3,HCVVOkamoto[9]Mori[10]2,3,ChanSim2mondsHCV,PCR(type-specificprimersPCR)[9],(restrictionfragmentlengthpolymor2phism,RFLP)[4],(lineprobeassay,Li2PA)[7,11],Livache[12]Bidan[13]HCV1HCVTable1ThetypespecificprobesofHCVNameSequenceTd()Length(bp)Typediscription15-ATTTGGGCGTGCC-3521813A1125-CAAATCTCCAGGCAT-34812141(1a/c,1b),635-CCCGCAAGACTGC-34914131a/c,645-CAGTCTCGCGGG-34513121b55-AAGGACCCAGTCTTCC-35210152(2a/c,2b)65-CAGTCTTCCCGGC-34719132a/c75-GAATTACCGGAAAGAC-35015162b85-CGAGTAGCGTTGGG-34919142(2a/c,2b),695-GCTTTCCGCGTGA-35015133(3a,3b)105-AAAGCGCCTAGCC-34912133a115-AAATTTCCGGGCAT-35117143bDNA,HCV,,GeneBank171HCV5NCR,Megalign(DNAstar)HCVHCV,,HCV5NCRCNS5E2/NS15NCR,,[1],5NCR,(1)19320039CHINESEJOURNALOFVIROLOGYVol.19No.3Sept12003©1995-2005TsinghuaTongfangOpticalDiscCo.,Ltd.Allrightsreserved.()50l,,-PCR,M-MLV(Promega)RNAPCRDNA,1:5-GGGCGACACTCCACCA-3;5-TGAGGTTTAGGATTCGTGCT-3:5-TCACTCCCCTGT2GAGGAA-3;5-AGCACCCTATCAGGCAGTA-3DNA260bp,PCR,dUTP(Biotin-11-dUTP,)dTTP13DNA,Roger[14],(),DNA1SSC9410min,4,,PCR1l,3523h,21PCRHCV5NCRDNAFigure1HCV5NCRDNAfragmentamplifiedbynested-PCRMW1DL2000(2,000bp,1,000bp,750bp,500bp,250bp,100bpfromtoptothebottom);1-71Resultsofpositivesera,4wasthesampletobetested;81Positivecontrol;91Negativecontrol12Figure2ResultstestedbyoligonucleotidearrayDNAarraysrepresentedrespectivelythespecificfragment:thegenotype1,subtypes1a/1c,1b;thegenotype2,subtypes2a/2c,2b;thegenotype3,subtypes3a,3b;thegenotype6andthenegativecontrol,allweretestedtwice111ArrayhybridizationresultsofHCVpositivesample;21ArrayhybridizationresultsofHCVgenotype3;31ArrayhybridizationresultsofHCVsubtype3b17,51,1b;13,3b;1143,PCRDNA,170(1)(3)3171HCV5NCR()Figure3PhylogenetictreeofpartialsequencesofHCV5NCRDNAfragmentsof171HCVcasesfromGeneBank1911004161P11seqisderivedfromse2quencing15723:3b©1995-2005TsinghuaTongfangOpticalDiscCo.,Ltd.Allrightsreserved.,4HCV3b(D4937D26556)HCV,113%113%,373HCV5NCR(4),HCV3b,4373HCV5NCRFigure4PhylogenetictreeofHCV5NCRDNAfrag2mentsofgenotype3of37HCVcasesandthederivedsequenceHCV,HCV,HCV33b,:[1]ChanSW,SimmondsP,McOishF,etal1SerologicalreactivityofblooddonorsinfectedwiththreedifferenttypesofhepatitisCvirus[J]1Lancet,1991,338:13911[2]ChanSW,McOmishF,HolmesEC,etal1Analysisofanewhepati2tisCvirustypeanditsphylogeneticrelationshiptoexistingvariants[J]1JGenVirol,1992,73:1131-11411[3]ChanSW,HolmesEC,McOmishF,etal1Phylogeneticanalysisofanew,highlydivergentHCVtype(type3):effectofsequencevariabil2ityonserologicalresponsestoinfection[A]1Venice:HepatitisCVirusandRelatedVirus:MolecularVirologyandPathogenesis[C].1992,D5:721[4]SimmondsE,McOmishE,YapPL,etal1Sequencevariabilityinthe5noncodingregionofhepatitisCvirus:Identificationofanewvirustypeandrestrictionsonsequencediversity[J]1JGenVirol,1993,74:661-6681[5]SimmondsP,AlbertiA,AlterHJ,etal1AProposedsystemforthenomenclatureofhepatitisCviralgenotypes[J]1Hepatology,1994,19:1321-13241[6]HoughtonM,WeinerA,HanJ,etal1Molecularbiologyofthehep2atitisCviruses:implicationsfordiagnosis1Developmentandcontrolofviraldisease[J]1Hepatology,1991,14:381-1881[7]EnomotoN,TakadaA,NakaoT,etal1TherearetwomajortypesofhepatitisCvirusinJapan[J]1BiochemBiophysResComm,1990,170:1021-10251[8]ChaTA,BeallE,IrvineB,etal1Atleastfiverelated,butdistinct,hepatitisCviralgenotypesexist[J]1ProcNatlAcadSciUSA,1992,89:7144-71481[9]OkamotoH,SugiyamaY,OkadaS,etal1TypinghepatitisCvirusbypolymerasechainreactionwithtype-specificprimers:Applicationtoclinicalsurveysandtracinginfectioussources[J]1JGenVirol,1992,73:673-6791[10]MoriS,KatoN,YangyuA,etal1AnewtypeofhepatitisCvirusinpatientsinThailand[J]1BiochemBiophysResComm,1992,183:334-3421[11]StuyverL,WyseurA,ArnhemW,etal1SecondgenerationlineprobeassayforhepatitisCvirusgenotyping[J]1JClinMicrobiol,1996,34(9):2259-22661[12]LivacheT,FouqueB,RogetA,etal1PolypyrroleDNAchiponasil2icondevice:exampleofhepatitisCvirusgenotyping[J]1AnalBiochem,1998,255(2):188-1941[13]BidanG,BillonM,GalassoK,etal1Electropolymerizationasaver2satilerouteforimmobilizingbiologicalspeciesontosur2faces1ApplicationtoDNAbiochips[J]1ApplBiochemBiotechnol,2000,89(2-3):183-1931[14]RogersYH,Jiang-BaucomP,HuangZJ,etal1Immobilizationofoligonucleotidesontoaglasssupportviadisulfidebonds:AmethodforpreparationofDNAmicroarrays[J]1AnalBiochem,1999,266:23-30167219©1995-2005TsinghuaTongfangOpticalDiscCo.,Ltd.Allrightsreserved.AGenotype3,Subtype3bofHepatitisCVirusFoundinWuhanbyUsingGeneChipWENJi-kai1,ZHANGXiang-ling2,ZHOUYa-feng1,WANGJian-chun2,ZHANGZhi-ping1ZHANGXian-en1(11WuhanInstituteofVirology,ChineseAcademyofSciences;Wuhan430071,China;21AssociateHospitalofWuhanIronandSteel(Group)Corporation;Wuhan430080,China)Abstract:1band2aHCVsubtypesweremainlyreportedinChina1Herein,a3bsubtypestrainofHCVwasfoundwithgenechipmethod1Firstly,the5NCR(non-codingregion)was