Dynamics of Hepatitis B Virus Quasispecies in Association with Nucleos(t)ide Analogue Treatment Determined by Ultra-Deep Sequencing 英文参考文献.docVIP

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Dynamics of Hepatitis B Virus Quasispecies in Association with Nucleos(t)ide Analogue Treatment Determined by Ultra-Deep Sequencing 英文参考文献.doc

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Dynamics of Hepatitis B Virus Quasispecies in Association with Nucleos(t)ide Analogue Treatment Determined by Ultra-Deep Sequencing 英文参考文献

DynamicsofHepatitisBVirusQuasispeciesin AssociationwithNucleos(t)ideAnalogueTreatment DeterminedbyUltra-DeepSequencing NorihiroNishijima1,HiroyukiMarusawa1*,YoshihideUeda1,KenTakahashi1,AkihiroNasu1, YukioOsaki2,TadayukiKou3,ShujiroYazumi3,TakeshiFujiwara4,SokenTsuchiya4,KazuharuShimizu4, ShinjiUemoto5,TsutomuChiba1 1DepartmentofGastroenterologyandHepatology,GraduateSchoolofMedicine,KyotoUniversity,Kyoto,Japan,2DepartmentofGastroenterologyandHepatology, OsakaRedCrossHospital,Osaka,Japan,3DepartmentofGastroenterologyandHepatology,TazukeKofukaiMedicalResearchInstitute,KitanoHospital,Osaka,Japan, 4Department ofNanobioDrug Discovery, Graduate School ofPharmaceutical Sciences,Kyoto University, Kyoto, Japan, 5DepartmentofSurgery,Graduate School of Medicine,KyotoUniversity,Kyoto,Japan Abstract Background and Aims: Although the advent of ultra-deep sequencing technology allows for the analysis of heretofore- undetectableminorviralmutants,alimitedamountofinformationiscurrentlyavailableregardingtheclinicalimplications ofhepatitisBvirus(HBV)genomicheterogeneity. Methods: To characterize theHBV genetic heterogeneity in association withanti-viral therapy, weperformed ultra-deep sequencingoffull-genomeHBVintheliverandserumof19patientswithchronicviralinfection,including14therapy-na?¨ve and5nucleos(t)ideanalogue(NA)-treatedcases. Results: Most genomic changes observed in viral variants were single base substitutions and were widely distributed throughouttheHBVgenome.Fourofeight(50%)chronictherapy-na?¨veHBeAg-negativepatientsshowedarelativelylow prevalenceoftheG1896Apre-core(pre-C)mutantinthelivertissues,suggestingthatothermutationswereinvolvedin theirHBeAgseroconversion.Interestingly,livertissuesin4of5(80%)ofthechronicNA-treatedanti-HBe-positivecaseshad extremely low levels of the G1896A pre-C mutant (0.0%, 0.0%, 0.1%, and 1.1%), suggesting the high sensitivity of the G1896Apre-CmutanttoNA.Moreover,variousabundancesofclonesresistanttoNAwerecommoninboththeliverand serumoftreatmen