适配体传感器在微生物检测中的应用

、、、.,,、、、、PCR(polymerasechainreaction)、.,,1～3d.PCR、、、、,,..:2011-05-05;:2011-07-29:(30770570):(1970-),,,,2010,,E-mail:[e-mail];*:(1960-),,,,,,Tel:[phone],E-mail:[e-mail].1,2,2*(1.,554300;2.,410081):,、、.SELEX(systematicevolutionofligandsbyexponentialenrichment)().,.,.、、,.、SELEX,.:;SELEX;;:Q93-332:A:1007-7847(2011)05-0455-05··ApplicationsofAptasensorsinDetectingMicroorganismWENXiao-tang1,LIHua2,DENGLe2*(1.TongrenPolytechnicCollege,Tongren554300,Guizhou,China;2.CollegeofLifeSciences,HunanNormalUniversity,Changsha410081,Hunan,China)Abstract:Aptamersarenucleicacidsequencesthatcanbindtoawidevarietyoftargetsfromsmallmoleculestowholecellswithhighaffinityandspecificity.Theycanbeobtainedinvitrothroughsystematicevolutionofligandsbyexponentialenrichment(SELEX)procedure.Aptamerspresentanalternativetoanti-bodiesasrecognitionagentsindiagnosticanddetectionassays.Aptamer-basedbiosensors(aptasensors)areanalysisdevices,whichcombinebiologicalrecognitioncomponentswithsignalconversionelementtodetecttargets.Providingrealtime,highsensitiveandspecifictests,aptasensortechnologyisoneofthefastgrowingareasinmicrobialrapiddetection.Advancesinaptamers,SELEXprocedures,aptasensorsandtheirapplica－tionsindetectionofmicroorganismarereviewed.Keywords:aptasensors;SELEX;detection;microorganismLifeScienceResearch2011155455～459155Vol．15No．5201110LifeScienceResearchOct.20112011,,.,,.,.,.,,[1].(RNA),RNA.2[2].,.(aptamersensor),.、[3].,(、)、,,,.,、[4].1SELEX(aptamer),aptus,“”,SELEX,.,、、、.,.SELEX、、、、,SELEX.SELEX,,,.1990TuerkGold[5,6],(systematicevolutionofligandsbyexponentialenrichment,SELEX),、.SELEX,PCR,,8～15,,、,,、,,.2,.、、、,,.、,.2.1,..Minunni[7]RNA,(HIV-1)Tat.、.Tat,0.25mg/L.,(QCM)(SPR),、.SPR(1～2.5mg/L)QCM(0～1.25mg/L).Lee[8](HCV).96,,Cy3,4565HCV.Gergely[9](PSA-H,MT32)DNA,.,.,.5mmol/L,.,,25%,3,SPR.,.2.2(QDs)(CNTs).QDs[10].QDs,.QDs-,.103cfu/mL,(6000cfu).105cfu/mL,.[11]SELEX,,5,.,,.John[12]DNA..,,.,15～20min.2.5cfu,10～250cfu.,,,,.Ohk[13]-A,,.,647.,P66A8.,,103cfu/mL.,O157∶H7,PT1,.,4cfu/g.、、.So[14](SWNT-FET)DH5α.105～107cfu/mL,SWNT-FET50%.,,.20min,DH5α、、.Gustavo[15],,..CECT675,O157∶H7.,,6(cfu),26cfu/mL.104cfu/mL,.,,48h.[16],4572011.(PDA),(LPS)PDA(CR).105cfu/mL,105～108cfu/mL,106cfu/mL6.08％,O111100％,30min.,、、、、,.[17]16SrDNA.525nm(QD)DNA1--3-(3-)(EDC)QD-DNA、6-(ROX)DNA16SrDNA.QDROX、.DNA,200nmol/L.,RNA、、;、(BHQ)-1.,-1tst,(530nm),BHQ(530nm)DNA,QD.QDtst,0.2μmol/L.、、.,.,、、、,,.3,,、、.,,.1),.,.、,,;2),..SELEXPCR,.,,.,,、、,.(References):[1]TORRES-CHAVOLLAE,ALOCILJAEC.Aptasensorsforde-tectionofmicrobialandviralpathogens[J].BiosensorsandBio-electronics,2009,24(11):3175-3182.[2]EULBERGD,KLUSSMANNS.Spiegelmers:biostableap-tamers[J].Chembiochem,2003,4(10):979-983.[3],,.[J].(ZHENGJing,HEPin-gang,FANGYu-zhi.Aptasensors[J].ProgressinChemistry),2009,21(4):732-738.[4],,,.[J].(YANGHai-peng,CHENShi-guo,LIChun-hui,etal.Nanomaterialsbasedelectrochemicalbiosensors[J].ProgressinChemistry),2009,21(1):211-216.[5]TUERKC,GOLDL.Systematicevolutionofligandsbyexpo-nentialenrichment:RNAligandstobacteriophageT4DNApolymerase[J].Science,1990,249(4968):505-510.[6]JAYASENASD.Aptamers:anemergingclassofmoleculesthatrivalantibodiesindiagnostics[J].ClinicalChemistry,1999,45(9):1628-1650.[7]MINUNNIM,TOMBELLIS,GULLOTTOA,etal.Develop-mentofbiosensorswithaptamersasbio-recognitionelement:4585ThecaseofHIV-1tatprotein[J].BiosensorsandBioelectronics,2004,20(6):1149-1156.[8]LEES,KIMYS,JOM,etal.Chip-baseddetectionofhepati-tisCvirususingRNAaptamersthatspecificallybindtoHCVcoreantigen[J].BiochemicalandBiophysicalResearchCommu-nications,2007,358(1):47-52.[9]GERGELYL,ZS魷FIAB,VIOLAB,etal.Aptamer-basedbiochipsforlabel-freedetectionofplantviruscoatproteinsbySPRimaging[J].TheAnalyst,2010,135(5):918-926.[10]IKANOVICM,RUDZINSKIW,BRUNOJ,etal.Fluorescenceassaybasedonaptamer-quantumdotbindingtoBacillusthuringiensisspores[J].JournalofFluorescence,2007,17(2):193-199.[11]CAOXX,LISH,CHENLC,etal.CombininguseofapanelofssDNAaptamerinthedetectionofstaphylococcus[J].NucleicAcidsResearch,2009,37(14):4621-4628.[12]JOHNGB,TAYLORP,MARIAPC,etal.Plastic-adherentDNAaptamer-magneticbeadandquantumdotsandwichassayforcampylobacterdetection[J].JournalofFluorescence,2009,19(3):427-435.[13]OHKSH,KOOOK,SENT,etal.Antibody-aptamerfunc-tionalizedfibre-opticbiosensorforspecificdetectionofListe-riamonocytogenesfromfood[J].JournalofAppliedMicrobiol-ogy,2010,109(3):808-817.[14]SOHM,PARKDM,JEONEK,etal.Detectionandtiteres-timationofEscherichiacoliusingaptamer-functionalizedsin-gle-walledcarbon-nanotubefield-effecttransistors[J].Small(WeinheimanderBergstrasse,Germany),2008,4(2):197-201.[15]ZELADA-GUILL魪NGA,BHOSALESV,RIUJ,etal.Real-timepotentiometricdetectionofbacteriaincomplexsamples[J].AnalyticalChemistry,2010,82(22):9254-9260.[16],,,.[J].(WUWen-he,CHENyong,JIANGLu-qian,etal.Anaptamerbasedbiosen－sorforcolorimetricdetectionofEnteropathogenicEscherichiacoli[J].ChineseJournalofLaboratoryMedicine),2010,33(7):587-593.[17]JIANGJF,PENGZH,DENGL,etal.Detectionofbifi-dobacteriumspecies-specific16SrDNAbasedonQDFRETbioprobe[J].JournalofFluorescence,2010,2