乙型肝炎病毒拉米夫定耐药突变及其基因型的研究

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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