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:1007-4287(2005)03-0477-041,1,2(1.,100730;2.),TRAP,RT2PCR,,,1(Telomereextensionassay)Morin[1]1989,:-70S2100,RNA,RNA,RNA,(TTAGGG)3,TTAGGG,2mMdATP,2mMdTTP,3mMa232PdGTP,30,60,011mgPmlRNARNARNA,,,,,,;,,,,,2(TRAP)Kim[2]1994,PCR:1)S2100;2):dNTPsTS(52AATCCGTCGAGCAGAGGT23),TS18bp,TS,RNA(hTP),TS3TTAGGG;3)PCR:,CX(52CCCTTACCCTTACCCTTACCCTAA23)Taq,TS,CXPCR6bp,PCR[32P]2dATP,15%,,,[3]PCR,10310,:1);2)Taq,3TRAPTRAP,Taq,:,;311TRAPWright[4]1995,150bp,TSCX,PCR,53TSCXTRAP,,186bp,,PCR,3122(TRAP2SPA)ErickaSavoyshy[5]TRAP(Scintillationproximityassay,SPA),TRAP2SPAKimTRAP:CX5(Bio2CX),dATPdCTPdGTP50MdTTP2M,2Ci[Me23H][232P]dGTP,T4g32,(40l)(50l)3710min,3H,,2h,,:,96,313Ohyashiki[6]TS,,8%,DNA,(fragmentmanagersystem),,90min,,1997OhyashikiTRAP,31[S]UTPRNA,314TRAP2ELISA1996,(),TS5,,7742005693PCR,,,PCR,,TMD,,TRAP,PCR,,68h[7]TRAP2ELISA,,22,,4(hybridizationprotectionassay,HPA)DavidBruceLackey[8]1998(HPA),,,,(NaOH),,,,H2O2,,,,PCR,TRAP,,,1h,Taq,:a1(TRAP)b1,dNTP,(TTAGGG)2,(),c1(AE)(52AA(CCCTTA)423),,(TTAGGG)4,(TTAGGG)8(TTAGGG)42(NaOH),H2O2d1(TTAGGG)4HPA,P,:30,301fmol(TTAGGG)41U5P(Transcription2mediatedam2plificationandhybridizationprotectionassay,TMAPHPA)1998Hirose[9]:(TMA)HPAHPAa1:,TRAP:,,3TTAGGG,T7RNA,T7RNA,;b1DNA,RTaseDNA;c1T7RNADNA,,RNA;d1RNA,,cDNA,DNAPRNA,RNaseH,RNADNA,,DNA;e1DNA1001000RNA100,;f1HPA:,HPA,HPAHPATMAPHPA,,GEN2PROBE,,,,,6PCRhTRhTERTmRNA3,RNA(hTR),(TP1)(hTERT),hTR,;TP1,Kyo[10]35,5,1112,hTERT,hTRTP1,hTERT,hTRTP1hTERTSumida[11]26hTERT,hTERTRT2PCRTRAPTchirkov[12],hTERTmRNAGSM,[1315]hTRhTERTPCRPCR1996AppliedBiosys2temsPCR,PCR,PCR,PCRSYBR,(TaqMan)611hTRYajima[16]TapManhTRPCR,874ChinJLabDiagn,June,2005,Vol9,No.3,,PCR,Taq5233,,,,DNA,,PCRhTR612hTERTmRNAYi[17]Schrader[1821]hTERTmRNA,RocheLight2CyclerTeloTAGGGhTERTQuantificationKit:RT2PCR,hTERTmRNA,,198bpcDNA,,,3,5LightCycler2Red640,3,DNA,,(FRET)FRET,LightCycler,LightCycler2Red640,,LightCyclerhTERTmRNA,,45LightCyclerTeloTAGGGhTERTQuantificationKit,10mRNA,102106,,,Morin1989,14,10,,TRAP,RT2PCR(hTERT)mRNA,,,,,,,:[1]MakarovVL,HiroseY,LangmoreJP.LongGtailsatbothendsofhu2manchromosomessuggestaCstanddegradationmechanismfortelomereshortening[J].Cell,1997,88:657.[2]KimNW,PlatyszekMA,ProwseKR,etal.Specificassociationofhu2mantelomeraseactivitywithimmotalcellsandcancer[J].Science,1994,266:2011.[3]WenJM,SunLB,ZhangM,etal.Anon2isotopicmethodforthedetec2tionoftelomeraseactivityintumortissues:TRAP2Silverstainingassay[J].JClinPathol:MolPathol,1998,51:110.[4]WrightWE,ShayJW,PiatyszekMA.Modificationsofatelomericrepeatamplificationprotocol(TRAP)resultinincreasedreliability,linearityandsensitivity[J].NucleicAcidsResrarch,1995,23:3794.[5]ErickaSavoyshy,Ken2ichiAkamatsu,MasayukiTsuchiya,etal.DetectionoftelomeraseactivitybycombinationofTRAPmethodandscintillationproximityassay(SPA)[J].NucleicAcidsResearch,1996,6(24):1175.[6]OhyashikiJH,OhyahikiK,SanoT.Non2radioisotopicandsemi2quanti2tativeprocedureforterminalrepeatamplificationprotocol[J].JpnJCan2cerRes,1996,87:329.[7],,,.2[J].,1999,22:24.[8]DavidBruceLachey.AHomogeneousChemiluminescentAssayforTelom2erase[J].Analyticalbiochemistry,1998,263:57.[9]MinoruHirose,JunkoAbe2Hashimoto,HidetoshiTahara,etal.Newmethodtomeasuretelomeraseactivitybytranscription2mediatedamplifica2tionandhybridizationprotectionassay[J].ClinicalChemistry,1998,44:2446.[10]SatoruKyo,TaroKanaya,MasahiroTakakuba.Expressionofhumante2lomerasesubuniteinovarianmalignant,borderlineandbeningtrmur[J].IntJCanaer,1999,80:804.[11]TomokiSumida,HiroyukiHamakawa,KenichiSogawa,etal.Telomer2asecomponentsasadiagnostictoolinhumanorallesions[J].IntJCan2cer,1999,80:1.[12]TchirhovA,RolhionC,KemenyJL,etal.Clinicalimplicationsofquantitativereal2timeRT2PCRanalysisofhTERTgeneexpressioninhu2mangliomas[J].BrJCancer,2003,88(4):516.[13]ParadisV,BiecheI,DargereD,etal.hTERTexpressioninsporadicrenalcellcarcinomas[J].JPathol,2001,195(2):209.[14]DeKokJB,SchalkenJA,AalersTW,etal.Quantitativemeasurementoftelomerasereversetranscriptase(hTERT)mRNAinurothelialcellcar2cinomas[J].IntJCancer,2000,15(2):217.[15]LiouMJ,ChanEC,LinJD,etal.Humantelomerasereversetran2scriptase(hTERT)mRNAgeneexpressioninFNAsamplesfromthyroidneoplasms[J].CancerLett,2003,19(2):223.[16]YajimaT,YagihashiA,KameshimaH,etal.Quantitativereversetranscription2PCRassayoftheRNAcomponentofhumantelomeraseus2ingtheTaqManfluorogenicdetectionsystem[J].ClinicalChemistry,1998,44(12):2441.[17]XiaomingYi,JerryWShay,WoodringEWright.Quantitationoftelom2erasecomponentsandhTERTmRNAsplicingpatternsinimmortalhumancells[J].NucleicAcidResearch,2001,29:4818.[18]SchraderM,MullerM,HeicappellR.etal.QuantificationofhumantelomeraseRNA(hTR)andhumantelomerasereversetranscriptase(hTERT)mRNAintesticulartissueofinfertilepatients[J].AsianJAndrol,2001,3:263.[19]SchraderM,BurgerAM,MullerM,etal.Quantificationofhumante2lomerasetranstriptasemRNAintesticulargermcelltumorsbyquantita2tivefluorescenceteal2timeRT2PCR[J].OncolRep,2002,9(5):1097.[20]SchraderM,MullerM,SchulzeW,etal.Quantificationoftelomerase9742005693activity,porphobilinogendeaminaseandhumantelomerasereversetran2scriptasemRNAintesticulartissue2newpa