二代测序技术与临床应用-高超

蛐,,··【】。LifeTechIonTorrent。。【】ClinicalApplicationoftheNext-GenerationSequencingTechnologyGAOChao，SHENJian-dong，CUIYu-gui.CenterofReproductiveMedicine，theFirstAffiliatedHospitalofNanjingMedicalUniversity，Nanjing210029，ChinaCorrespondingauthor：CUIYu-gui，E-mail：[e-mail]【Abstract】Inrecentyears,thesequencingtechnologyhasbeeninnovatedfollowingthebreakthroughoftheengineeringmaterialbottleneck.Thehigh-throughputgene-sequencingtechnologymaturedgraduallymadethetransformationfromroutineandbasicresearchlaboratorytoclinicalpractice,becausethenext-generationsequencing(NGS)technologymettheneedsofindividualdiagnosisandtreatment.Inthispaper,thedevelopingprogressofNGStechnology,andoperationalprocess,wereintroduced.Thosemainstreamproductsintheinternationalmarketatpresentwerealsointroduced.TheIonTorrentsequencingplatformofLifeTechCompanywasintroducedindetail,includingitsbasicprinciple,laboratoryprotocol,coefficientofperformanceandclinicalapplication.ThispaperalsoprovidedsomeexamplestoshowthespecificsignificanceinclinicalapplicationandpotentialinsufficientofNGStechnology.【Keywords】Sequenceanalysis；Moleculardiagnostictechniques；Genetictechniques（JIntReprodHealth/FamPlan，201433154-156171）[e-mail]20SangerDNA21、next-generationsequencing,NGSDNA、DNA、。DNA“”。IlluminaLifeTech。LifeTechIonTorrentIonTorrentIonTorrent。NGS。1NGS454LifeSciences454Roche2005NGS2006SequencingbySynthesisSolexa1992LynxTherapeuticsMPSSSolexaIllumina154··蛐,,2003GeorgeChurchPolonyABISOLiD2007。[1]。DNA2010DNADNANGS。454RothbergJM2007IonTorrentABILifeTech2010IlluminaIonTorrentDNA。2IonTorrent2.1IonTorrentDNA1DNA5′3′1。DNA。。IonTorrent。。IonTorrent。IonTorrentSangerDNAhomopolymersIonTorrent。2.2IonTorrentPersonalGenomeMachinePGM10GProton。DNARNAIonTorrentDNAPCRPCRwater-in-oilemulsionPCR[2-4]。DNADNApolyclones。155··蛐,,DNA。2.3IonTorrentPGM10M、100M、1G314、316、318DNAIonTorrent。IonTorrentIonTorrent。3NGSSanger。NGS。IlluminaMiSeqLifeTechIonTorrent。3.1NGS。NGS[5][6-7]。NGS[8]。NGS。3.2NGS。NGS。3.3NGSRNARNA。DNANGSDNA。4、。NGS。。p171156··蛐,,。NGSNGS。DNA。[1]GlennTC.Fieldguidetonext-generationDNAsequencers[J].MolEcolResour2011115759-769.[2]ShaoKDingWWangFetal.EmulsionPCR:ahighefficientwayofPCRamplificationofrandomDNAlibrariesinaptamerselection[J].PLoSOne201169e24910.[3]WilliamsRPeisajovichSGMillerOJetal.AmplificationofcomplexgenelibrariesbyemulsionPCR[J].NatMethods200637545-550.[4]XuMYAragonADMascarenasMRetal.DualprimeremulsionPCRfornext-generationDNAsequencing[J].Biotechniques2010485409-412.[5]MorrissyASMorinRDDelaneyAetal．Next-generationtagse－quencingforcancergeneexpressionprofiling[J]．GenomeRes200919101825-1835．[6]PanQShaiOLeeLJetal．Deepsurveyingofalternativesplicingcomplexityinthehumantranscriptomebyhigh-throughputsequencing[J]．NatGenet200840121413-1415．[7]PfiferGPBesaratiniaA．Mutationalspectraofhumancancer[J].HumGenet20091255/6493-506．[8]FanHCBlunebtekdYJChitkaraUetal．NoninvasivediagnosisoffetalaneuploidybyshotgunsequencingDNAfrommaternalblood[J]．ProcNatlAcadSciUSA20081054216266-16271．2014-04-09[]p156[1].[M].7.2008.[2]SuzumoriNSugiura-OgasawaraM.Geneticfactorsasacauseofmiscarriage[J].CurrMedChem201017293431-3437.[3].139[J].198382129-131.[4].2720[J].2006163173-174.[5].1780[J].2005303258-260.[6].1016[J].200924101374-1377.[7]DuttaURRajithaPPiduguVKetal.Cytogeneticabnormalitiesin1162coupleswithrecurrentmiscarriagesinsouthernregionofIndiareportandreview[J].JAssistReprodGenet2011282145-149.[8]EibenBBartelsIB覿hr-PorschSetal.Cytogeneticanalysisof750spontaneousabortionswiththedirect-preparationmethodofchorionicvillianditsimplicationsforstudyinggeneticcausesofpregnancywastage[J].AmJHumGenet1990474656-663.[9]McAuliffeMEWilliamsPLKorrickSAetal.Theassociationbetweenspermsexchromosomedisomyandsemenconcentrationmotilityandmorphology[J].HumReprod201227102918-2926.[10]Sugiura-OgasawaraMOzakiYKatanoKetal.Abnormalembryonickaryotypeisthemostfrequentcauseofrecurrentmiscarriage[J].HumReprod20122782297-2303.[11]GhazaeySMirzaeiFAhadianMetal.PatternofchromosomalaberrationsinpatientsfromnorthEastiran[J].CellJ2013153258-265.[12]TeoSHTanMKnightLetal.Pericentricinversion9--incidenceandclinicalsignificance[J].AnnAcadMedSingapore1995242302-304.[13]CaglayanAOOzyazganIDemiryilmazFetal.Areheterochromatinpolymorphismsassociatedwithrecurrentmiscarriage?[J].JObstetGynaecolRes2010364774-776.[14].[J].200729111362-1366.[15]Akba鬤HIsiHOralDetal.Chromosomeheteromorphismsaremorefrequentincoupleswithrecurrentabortions[J].GenetMolRes20121143847-3851.2014-02-19[]171··