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ID:20866674
大小:11.22 MB
页数:89页
时间:2018-10-17
《乙型肝炎病毒拉米夫定耐药突变及其基因型的研究》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、乙型肝炎病毒拉米夫定耐药突变及其基因型的研究ahighdegreeofautomation,andituseslesstimeforhybridization,therefore,itissuitaleforlarge—scaleHBVgenetyping,However,therearemanyfactorsintheprocessofthiSmethod,anditsoperationrequireacertaintechnique,inadditiontothenormalreactioninclini
2、calexamination,therearealwayssomeerrorswecanseeresults(falsepositiveorfalsenegativeresults).Inaddition,inrecentyearsthedevelopmentofgenechiptechnologytodetectHBVgenotypebecomesmoresimple,efficientandaccurateaspossible,therearemanypotentialadvantagesforthiSm
3、ethodgenotypingHBV,butintherealizationprocesstherearemanyissuestoberesolved.ObjectiveTooptimizethreemethodsofRFLP,quantitativePCRanddenaturinghigh—performanceliquidchromatography(DHPLC)fordetectinglamivudineresistanthepatitisBvirusmutationandgenotypingHBV,t
4、ocompareandevaluatethedetectionresultsofthethreemethodsforgettingamethodwhichcandetectlamivudineresistantmutationsandhaveHBVgenotypingquickly,accuratelyandstability:alsoitisapplicabletoclinicallydetectlamivudineresistantmutationsinHBVandhaveHBVgenotypingina
5、largescale.Toinvestigatetherelationshipbetweenthelamivudineresistantmutations,genotypesofHBVandclinicaltherapybyanalyzingtheresultsofdetectionandclinicaldata.Methods1.usingRFLPmethodtodetectlamivudine—resistantmutationsinHBVandhaveHBVgenotyping:Firstly,desi
6、gnprimersusedfordetectingmutationsandgenotypingaccordingtotheanalysisofblastandreportedintheliterature,andthen,optimizethedigestionsystemforgettingthebestone:Finally,thepurifiedPCRproductswereconductedthroughelectrophoresis,anddeterminemutationsandgenotypes
7、ofHBVthrough7山东省医学科学院硕士论文observingthegelunderthegelimagingsystem.2.Real—timePCRwasusedtolamivudine—resistantmutationsandgenetypeofHBV:Firstly,designsevenpairsofprimersusedfordetectinghepatitisBvirusLamivudine——resistantmutationinaccordancewiththemutationpoi
8、ntinHBV;hepatitisBvirusisdividedintoeightkindsofgenotypes,ChinamainlyhaveB,C,Dthreegenotypes,SOdesignprimersdesignfordetectingthethreegenotypesofB,CandD:Then,optimizethereal-timefluorescencePCRreaction
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