滚环扩增原理

DNA肖乐义1*祁自柏2姜俊1张子宽3(145000321000503450003)DNA(rollingcircleDNAamplification,RCA),105,109,()DNARCA(on2chip),,RCA,DNAphi29DNA:2004203222:2004206228:[e-mail],,DNA(rollingcirclereplication,RCR)[1]RCA,()DNA,RCA(on2chip),,(microarray),,PCRT7[2],(),,,(off2chip),RCA1DNA,DNA,,,DNA,dNTPRCA109[1],105[3]RCA,DNA()DNA,DNA,DNA,dNTPDNA,DNA(1)RCA(multiply2primedRCA)DNADNARCA(2),RCA,DNA,,RCARCA()RCA,DNADNA(phi29DNARNAbstDNA[4~6]),DNADNAphi29DNA:(1)phi29DNA,;(2)DNA70kb,[7],mRNA,249CHINABIOTECHNOLOGY20049RNA[8]RCA,,DNADNAdNTP,DNADNA,DNA,(3)RCARCARCA,PCR(on2chipamplification),:(1);(2);(3),;(4)(off2chip),;(5)1Fig.1Schemeforgenerationofconcatemerusingrollingcirclereplication2DNA211RCA,,DNA,RCADNA,:DNA,RCAphi29DNA(wholegenomeamplification,WGA)DNAMDA30e4~6,,,DNA5logs(100fg~10ng),,100LlDNA20~30Lg,,DNA10kb,MDA10WGA3424,DNA[9]2Fig.2Multiply2primedrollingcircleamplification(multiply2primedRCA)3Fig.3Schematicrepresentationofimmuno2RCAapproach,MDAPCR10kb,,MDAphi29DNA106~107[10],TaqDNAPCR1043[11],,PCR20900[12]DNA,DNA,10000,MDADNA,(24),,,TempliphiDNAphi29DNA,,(4)43~5Lg4TempliphiDNA212DNADNA(padlockprobes)cDNA,DNARNA,WilsonATP7B[13]RCA,RCA21211液相RCAdNTPs,phi29DNA,80DNA20,25000phi29DNA,T7DNARCA,phi29DNA50ntPs,301000,T7DNA(300ntPs),T7DNA,359:DNAphi29DNA1021212玻片微阵列上RCA,,DNA,phi29DNA3,8125,521213多孔基质上RCAPEG2200,phi29DNA,1000;,phi29DNARCA1000,BstDNAT7DNARCA5~10[14],RCAPCR,213RCARCA(singlenucleotidepolymorphisms,SNPs)RCADNA,,SNPDNA,3c,DNA;,,DNA,,RCARCA,DNA,(5),,[15]RCASNP(L2RCASNP)PCR(~50ng)DNADNA,,,phi29DNADNACybr2gold,PCRRCA,3624[16]214RCAIgE,ELISAIgE011ngPml,RCAIgE,RCAIgEELISAPSA,RCA011pgPmlIgE,RCACy3,,5logs,1ngPmlIgE,RCA1pgPmlIgE,5logs[17]2,,,,,RCA,RCA75,femtomolar,3,RCA1000RCA[18]RCA2150,,,RCA215RCARCARCA,RCA,[1,19],21511一般局部化抗原的检测RCA,B(CD20),RCA,,,RCA,4CD20,EMA,,RCA421512免疫RCA用于流式细胞仪CD3RCARCA,CD3Jurkat40,CD360IgERCA(microspheres)(bead),,RCA,TheUpstateBiotechnologyAssay5,IL22IL24A2TheBiosourceInternationalAssay10,5IL21BIL25IL28IL210RCA,100,,1533[20]3RCA,RCA,SNPRCAPCR,,RCA,,,,RCA379:DNARCA,RCA-RCA,RCA[1]LizardiPM,HuangX,ZhuZ,etal.Mutationdetectionandsingle2moleculecountingusingisothermalrolling2circleamplification.NatGenet,1998,19(3):225~232[2]PhililpsJ,EberwineJH.AntisenseRNAamplification:AlinearamplificationmethodforanalyzingthemRNApopulationfromsinglelivingcells.Methods,1996,10(3):283~288[3]YuriyG,JasonS,ArumughamR,etal.Rollingcircleamplificationanewapproachtoincreasesensitivityforimmunohistochemistryandflowcytometry.AmericanJournalofPathology,2001,159:63~69[4]VoiseyJ,HafnerGJ,MorrisCP,etal.InterrogationofmultimericDNAamplificationproductsbycompetitiveprimerextensiveusingbstDNApolymerase(largefragment).Biotechniques,2001,31(5):1122~1124,1126,1128~1129[5]ZhangDY,ZhangW,LiX,etal.DetectionofrareDNAtargetsbyisothermalramificationamplification.Gene,2001,274(122):209~216[6]HafnerGJ,YangIC,WolterLC,etal.IsothermalamplificationandmultimerizationofDNAbyBstDNApolymerase.Biotechniques,2001,30(4):8522856,858,860[7]BlancoL,BernadA,LazaroJM,etal.HighlyefficientDNAsynthesisbythephagephi29DNApolymerase.SymmetricalmodeofDNAreplication.JBiolChem,1989,264(15):8935~8940,8905[8]ZhouY,CalcianoM,HamannS,etal.InsitudetectionofmessengerRNAusingdigoxigenin2labeledoligonucleotidesandrollingcircleamplification.ExpMolPathol,2001,70(3):281~288[9]FrankBD,SeiyuH,FangLH,etal.Comprehensivehumangenomeamplificationusingmultipledisplacementamplification.PNAS,2002,99(8):5261~5266[10]EstebanJA,SalasM,BlancoL.Fidelityofphi29DNApolymerase.Comparisonbetweenprotein2primedinitiationandDNApolymerization.JBiolChem,1993,268(4):2719~2726[11]EckertKA,KunKelTA.DNApolymerasefidelityandthepolymerasechainreaction.PCRMethodsAppl,1991,1(1):17~24[12]SaikiPK,GelfandDH,StoffelS,etal.Primer2directedenzymaticamplificationofDNAwithathermostableDNApolymerase.Science,1998,239(4839):487~491[13]JohanB,AndersI,ErikW,etal.UlfLandegrenandMatsNilssonparallelgeneanalysiswithallele2specificpadlockprobesandtagmicroarrays.NucleicAcidsResearch,2003,31(17):e107[14]GirishN,ChenghuaLuo,LinhuaFang,etal.SignalamplificationbyrollingcircleamplificationonDNAmicroarrays.NucleicAcidsResearch,2001,29(23):e118[15]JudithP,AnonaB,VarshaG,etal.IntegrationofDNAligationandrollingcircleamplificationforthehomogeneous,end2pointdetectionofsiglenucleotidepolymorphisms.NucleicAcidsResearch,2002,30(12):e60[16]XiaoquanQI,SalehaB,KatrienM,etal.L2RCA(ligation2rollingcircleamplification):ageneralmethodforgenotypingofsinglenucleotidepolymorphisms(SNPs).NucleicAcidsResearch,2001,29(22):e116[17]BarryS,StevenW,JeremyL,etal.ImmunoassayswithrollingcircleDNAamplification:Aversatileplatformforultrasensitiveantigendetection.PNAS,2000,97(18):10113~10119[18]SchweitzerB,RobertsS,GrimwadeB,etal.Multiplexedproteinprofilingonmicroarraysbyrolling2circleamplification.NatureBiotechnology,2002,20(4):359~365[19]XiaoboZhong,PaulML,XiaohuaHuang,etal.WardvisualizationofoligonucleotideprobesandpointmutationsininterphasenucleiandDNAfibersusingrollingcircleDNAamplification.PNAS,2001,98(7):3940~3945[20]MichaleCM,RamousS,WilliamJ,etal.Rollingcircleampli