【】分子生物学技术在临床检验中的应用PPT课件VIP

-,1,分子生物学技术在临床检验中的应用Molecularbiologytechniquesinaclinicallab,杨正林四川省医学科学院四川省人民医院,-,2,为什么要用分子生物学技术诊断人体疾病?(Whydoweneedmolecularbiologytechniquesforclinicaldiagnosis?),-,3,不同层次检验分析,分子水平(molecular)生化水平(biochemistry)细胞水平(cell)组织水平(tissue)个体水平(individual)群体水平(population),-,4,致病有机体,人体内部系统自身紊乱,（微生物寄生虫等）,（如：malaria）,原核,真核,胞膜、胞浆,胞核（染色体等）,蛋白、糖、脂核酸（DNA、RNA等）,分子水平,组织,胞膜、胞浆（包括线粒体等），胞核（染色体等）,蛋白、糖、脂核酸（DNA、RNA等-）,化学物、空气等,机体水平,(组织）细胞水平,亚细胞结构,细胞,人体疾病HumanDisease,microRNA,siRNA,外伤Trauma,-,5,什么是分子生物学？Whatdoesmolecularbiologymean?,Molecularbiologyisthestudyofbiologyatamolecularlevel.Thefieldoverlapswithotherareasofbiologyandchemistry,particularlygeneticsandbiochemistry.MolecularbiologychieflyconcernsitselfwithunderstandingtheinteractionsbetweenDNA,RNAandproteinbiosythesisandlearninghowtheseinteractionsareregulated,-,6,ExpressioncloningPolymerasechainreaction(PCR)GelelectrophoresisSouthernblottingNorthernblottingWesternblottingSequencingLinkageanlysisArraysAllelespecificoligonucleotideAbandonedtechnology,什么是分子生物学技术？Whataretechniquesofmolecularbiology?,-,7,分子生物学技术字在临床检验中的应用,1.感染性疾病中应用2.人类疾病相关基因的检查3.肿瘤的检测(differentiate)、预防和治疗（包括疗效观察）4.pharmacogenomics5.组织配型HLA2（DR、Q）6.亲子鉴定7.法医鉴定,-,8,-,9,ClevelandClinicalFoundation,-,10,ARUP,-,11,遗传性疾病的类型,1.Chromsomaldisease2.Single-genedisease,monogenicdiseaseAutosomaldominantinheritance,ADAutosomalrecessiveinheritance,ARX-linkeddominantinheritance,XDX-linkedrecessiveinheritance,XRY-linkedinheritance,-,12,3.Polygenicdisease,Multifactorialdisease(1)A.Majorgene+othergenes+otherfactorsB.Manyminorgene+otherfactors(2)Veryimportant,includingalotofcommondiseases(3)ResearchmethodsPopulationgenetics,STRlinkageanalysis,SNPs4.Mitochondrialgenicdisease,mtDNA5.Somaticcellgeneticsdisease,遗传性疾病的类型（续）,-,13,DNA分子结构改变-基因突变,1.Chromosomeaberration2.GenemutationPointmutationDeletionInsertionTandemrepeat,-,14,基因突变的类型,1.transitionortransversionsamesensemutationmissensemutationnon-sensemutationterminationcodonmutation2.Frameshiftmutationa(includingsomesplicingmutations)3.Condonmutation(Condoninsertionorinsertion)4.Mispairedsynapsisandunequalcrossing-over-Fusiongene5.Dynamicmutation,-,15,3.Condonmutation(Condoninsertionorinsertion)4.Mispairedsynapsisandunequalcrossing-over-Fusiongene5.Dynamicmutation,基因突变的类型(续）,-,16,疾病相关基因的检查,细胞遗传学（Cytogenetics）分子遗传学（MolecularGenetics）,-,17,细胞遗传学（Cytogenetics）,是遗传学的一个分支主要研究染色体和细胞分裂(Chromosomesandcelldivision)常用的分析技术包括G带染色(G-banded)等和分子细胞遗传学技术(MolecularCyrogenetics)如FISH(Fluorescentinsituhybridization)和CGH(Comparativegenomichybridization),-,18,Humanmalekaryotype,-,19,Ametaphasecellpositiveforthebcr/ablrearrangementusingFISH,-,20,Interphasecellspositiveforat(9;22)rearrangement,-,21,分子遗传学（Cytogenetics）,Moleculargeneticsisthefieldofbiologywhichstudiesthestructureandfunctionofgenesatamolecularlevel.Thefieldstudieshowthegenesaretransferredfromgenerationtogeneration.Moleculargeneticsemploysthemethodsofgeneticsandmolecularbiology.Itisso-calledtodifferentiateitfromothersubfieldsofgeneticssuchasecologicalgeneticsandpopulationgenetics.,-,22,Animportantareawithinmoleculargeneticsistheuseofmolecularinformationtodeterminethepatternsifdescent,andthereforethecorrectscientificclassificationoforganisms:thisiscalledmolecularsystematics.Alongwithdeterminingthepatternofdescendants,moleculargeneticshelpsinunderstandinggeneticsmutationsthatcancausecertaintypesofdisease.Throughutilizingthemethodsofgeneticsdiscoversthereseasonswhytraitsarecarriedonandhowandwhysomemaymutate.,分子遗传学（Cytogenetics）(续）,-,23,分子遗传学技术（Techniquesofmolecularbiology）,ForwardgeneticsReversegeneticsGeneTherapyTechniquesinmoleculargeneticsAmplification:PCRandcloningDNAinbacteriaSeparationanddetection:cellculture,DNAIsolation,mRNAisolationThemoleculargeneticsproject基因表达定量,-,24,检查突变的方法,SSCPDGGEDHPLCDigestionPrime3-ASODirectSequencingGenotyping(STR)Snapshot,-,25,-,26,Pharmacogenomics简介,-,27,疾病相关基因检查的质量控制,1.检查灵敏度2.检查特异性3.准确性4.质量控制,-,28,疾病相关基因检查的未来展望,-,29,遗传基因几乎与所有人类疾病相关,医学科学的发展使感染性疾病和营养性因素所致疾病的发病率越来越低，并被逐渐控制。医学工作者的注意力已转向遗传性疾病的防治。随着人类遗传学和医学遗传学的发展，人们已认识到遗传因素存在于除外伤以外几乎所有的