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1、TheHybridCapture2(hc2)System HPVDNATest by DigenebyBrendaPalaciosObjectivesStatethe2mainlow-riskHPVDNAtypesStatethe2mainhigh-riskHPVDNAtypesStatewhattypeoftestmethodisusedforthedetectionofHPVStatetheminimumsamplevolumerequiredfortestingStatewhatthemolecularsandwichconsistofHu
2、manPapillomavirus(HPV)Primaryetiologicalagentincervicalcancer2ndmostcommontypeofcancerinwomenworldwide3rdleadingcauseofcancer-relateddeathsinwomenworldwideMostcommonviralsexuallytransmittedinfection,thatgoesundiagnosedduetonosymptomsdevelopedHPVCharacteristicsNon-envelopeddou
3、ble-strandedDNAvirusEpitheliotrophic-hasgreataffinityforepithelialcellsObligatoryintracellularparasitesthatdelivertheirgenomeandaccessoryproteinsintohostcellsforviralreplicationHPVInfectionE6oncogenebindstop53proteininhostcellp53proteinisanegativeregulatorE6proteinmutatesp53p
4、roteinremovingitsprotectivefunctionMutationdisablesp53geneswitch,permittingcelltomultiplyuncontrolledHPVTypesTypes6&11–mostcommonlow-riskHPVAssociatedwithgenitalwartsRarelyfoundincervicalcancerTypes16&18-mostcommonhigh-riskHPVAssociatedwithcancersofthecervix,vagina,vulva,anus
5、,andpenisHPVDetectionTraditionalscreeningbyPapanicolau(papsmear)testUseduniversallyforinitialdetectionofintraepithelialabnormalitiesIncaseofatypicalsquamouscellsofundeterminedsignificanceHPVDNAdetectionisrecommendedwww.unsw.edu.au/.../sep/cells_image_inside.jpgMolecularTestin
6、gHybridCapture2(hc2)SystemHPVDNATestApprovedbyFDANucleicacidhybridizationassayNotargetDNAamplificationSingleamplificationusingmicroplatechemiluminescenceforqualitativedetectionof18typesofHPVwww.clpmag.com/graphics/mags/0409/sl03.jpgMolecularTestingcont.Differentiatesbetweenlo
7、wandhighriskHPVLow-riskHPV:6,11,42,43,44High-riskHPV:16,18,31,35,39,45,51,52,56,58,59,68DoesnotdeterminespecificHPVgenotypeControlsandReagentsControlsHigh-riskcontrol:clonedHPV16DNALow-riskcontrol:clonedHPV6DNANegativecontrolCarrierDNACalibrators(runintriplicate)Low-riskCalib
8、rator:ClonedHPV11DNAHigh-riskCalibrator:clonedHPV16DNAEnsurethatther
