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Real-TimeNucleicAcidSequence-BasedAmplificationIsMoreConvenientthanReal-TimePCRforQuantificationofPlasmodiumfalciparum
...interventionorvaccinetrials.Tocircumventthisproblem,twomoresensitiveassays,real-timequantitativenucleicacidsequence-basedamplification(QT-NASBA)andreal-timequantitativePCR(QT-PCR)werecomparedforquantificationofP.falciparumparasites.QT-NASBAwasadaptedtomolecularbeaconreal-timedetectiontechnology,whic...
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Real-Time-PCRAssayforDiagnosisofEntamoebahistolyticaInfection
...Pathology,UniversityofVirginia,Charlottesville,VirginiaABSTRACTWedevelopedareal-time-PCRassayutilizingamolecular-beaconprobeforthedetectionofEntamoebahistolyticaandcompareditssensitivitytostoolantigendetectionandtraditionalPCR.Atotalof205stoolandliverabscesspusspecimensfrompatientsandcontrolswereuse...
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DevelopmentofOne-Step,Real-Time,QuantitativeReverseTranscriptasePCRAssaysforAbsoluteQuantitationofHumanCoronavirusesOC43and229E
...clinicalsetting.Inthisstudy,wepresentaverysensitiveandspecificTaqMan-based,real-timequantitativereversetranscriptasePCR(qRT-PCR)fortherapiddetectionandquantitationofhumancoronaviruses(HCoVs)OC43and229E.Absoluteviralloadmeasurementinclinicalsampleswasachievedthroughtheconstructionofin-houseHCoVOC43an...
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RapidDetectionandDifferentiationofDengueVirusSerotypesbyaReal-TimeReverseTranscription-Loop-MediatedIsothermalAmplificationAssay
...ration,Saitama332-0012,JapanABSTRACTThedevelopmentandvalidationofaone-step,real-time,andquantitativedenguevirusserotype-specificreversetranscription-loop-mediatedisothermalamplification(RT-LAMP)assaytargetingthe3‘noncodingregionfortherapiddetectionanddifferentiationofdenguevirusserotypesarereporte...
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PerformanceofSingle-StepGel-BasedReverseTranscription-PCR(RT-PCR)AssaysEquivalenttoThatofReal-TimeRT-PCRAssaysforDetectionoftheSevereAcute
...me(SARS)outbreakinSingapore,werefoundtobeassensitiveascommercialandin-housereal-timeRT-PCRassays.Thedetectionlimitwasapproximately1genomeequivalent(GE)per5μlPCRmixture.OnePFUofSARScoronaviruswasestimatedtobe258±46GE.TEXTSevereacuterespiratorysyndrome(SARS)emergedin2002-2003asaglobaloutbreakcausedb...
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UseofMolecularBeaconsforRapid,Real-Time,QuantitativeMonitoringofCytotoxicT-LymphocyteEpitopeMutationsinSimianImmunodeficiencyVirus
...heyrepresentasfewas500mutantcopiesinasamplecontaining10,000totalcopies.Thisreal-timePCRtechniqueforevaluatingCTLepitopemutationsmayprovetobeausefultoolformonitoringthegeneticdriftofhumanimmunodeficiencyvirusandSIVininfectedindividuals.INTRODUCTIONCytotoxicTlymphocytes(CTL)playanimportantroleincontro...
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治疗性血管新生实验研究的若干进展
...对照心肌ABCDExpressionchangesofbFGFR1afterratacutemyocardialinfarction(Real-timeRT-PCR,n=5)*#*#*#*#*#*#****//GEArrayTMQseriesMouseAngiogenesisGeneArray小鼠血管新生基因芯片(SuperArray,U.S.A.)非梗死区边缘区心肌梗死大鼠心肌细胞内血管新生基因表达的变化(Micro-ar...
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ComparisonofThreeDifferentPCRMethodsforQuantifyingHumanPapillomavirusType16DNAinCervicalScrapeSpecimens
...athologyLaboratory,StichtingPAMM,Eindhoven,TheNetherlandsABSTRACTWecomparedreal-timeLightCyclerandTaqManassaysandtheGP5+/6+PCR/enzymeimmunoassay(EIA)toassessthehumanpapillomavirustype16(HPV16)loadincervicalscrapespecimens.Bothreal-timePCRassaysdeterminedtheHPV16loadinscrapespecimenssimilarly.Theleve...
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发现长期施肥对稻田土壤固碳自养菌多样性影响
...期定位试验为平台，采用PCR-克隆测序和实时荧光定量（Real-time）PCR技术，研究不施肥（CK），氮磷钾肥（NPK）和秸秆还田（NPKS）3种长期施肥制度对稻田土壤固碳自养菌群落结构及数量的影响。 通过分析固碳细菌cbbL基因文库...
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Real-TimeNucleicAcidSequence-BasedAmplificationUsingMolecularBeaconsforDetectionofEnterovirusRNAinClinicalSpecimens
...alVirologyLaboratory,YaleNewHavenHospital,NewHaven,Connecticut06504ABSTRACTReal-timenucleicacidsequence-basedamplification(NASBA)usingmolecularbeacontechnology(NASBA-beacon)wascomparedtostandardNASBAwithpostamplificationhybridizationusingelectrochemiluminescentlylabeledprobes(NASBA-ECL)fordetectiono...
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MaternalCocaineAdministrationCausesanEpigeneticModificationofProteinKinaseCGeneExpressioninFetalRatHeart
...eratioofparticulate/cytosolicPKCwasusedtodeterminetherelativeactivityofPKC.Real-TimeRT-PCR.RNAwasextractedfromheartsusingTRIzolreagents(Invitrogen,Carlsbad,CA).PKCmRNAlevelsweredeterminedbyreal-timeRT-PCRusinganIcyclerThermalcycler(Bio-RadLaboratories).SpecificPKCprimerswere5‘-GCGAAGCCCCTAAGACAAT-...
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miR21在白血病发病过程中的作用研究
...单碱基锚定OligdT引物进行反转录，然后用特异引物进行realtimePCR扩增，从而检测部分miRNAs在白血病细胞株K562中的表达，并与正常人骨髓表达情况相对比。结果通过realtimePCR分析，癌基因miR21在K562细胞中的表达量约为(1.73±...
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Real-TimePCRforDetectionandIdentificationofPlasmodiumspp.
...malariaparasitesinbloodisneededtoinstitutepropertherapy.Wedevelopedandusedareal-timePCRassaytodetectanddistinguishfourPlasmodiumspp.thatcausehumandiseasebyusingasingleamplificationreactionandmeltingcurveanalysis.Consensusprimerswereusedtoamplifyaspecies-specificregionofthemulticopy18SrRNAgene,andSYB...
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UseofBacteriophageMS2asanInternalControlinViralReverseTranscription-PCRAssays
...se-negativeresultsduetoinhibitionorhumanerror.Inthisstudy,wedesignedvariousreal-timeRT-PCRsutilizingthecoliphageMS2replicasegene,whichdifferindetectionformat,ampliconsize,andefficiencyofamplification.ThesenoncompetitiveICassays,usingTaqMan,hybridizationprobe,orduplexscorpionprobetechniques,wereteste...
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ComparisonofSmartCyclerReal-TimeReverseTranscription-PCRAssayinaPublicHealthLaboratorywithDirectImmunofluorescenceandCellCultureAssaysina
...tractIntroductionMaterialsandMethodsResultsDiscussionReferencesAsingle-tubereal-time(fluorogenic)reversetranscription(RT)-PCRwiththeSmartCyclerinstrument(SmartCyclerRT-PCR)forinfluenzaAvirusdetectionwasevaluatedwith238respiratoryspecimens.Directimmunofluorescenceantibodystaining(DFA)andprimaryrhesus...
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EvaluationoftheLightCyclerStaphylococcusMGRADEKitsonPositiveBloodCulturesThatContainedGram-PositiveCocciinClusters
...iinbloodculturebottlesandsimultaneousidentificationofS.aureusamongthemusingreal-timePCRhavealsobeendemonstrated(8).ThisapproachidentifiesCoNSbyactuallydetectingitspresenceratherthanbyassumingitspresenceintheabsenceofS.aureusinculturesgrowingGPCC.Alimitationtotheuseofreal-timePCRfordetectionofstaphyl...
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Real-TimeReverseTranscriptionPCRforDetectionandQuantitativeAnalysisofEquineInfluenzaVirus
...zooticrespiratorydiseaseoftheequine.Thedetectionofequineinfluenzavirususingreal-timeLightCyclerreversetranscription(RT)-PCRtechnologywasevaluatedovertwoinfluenzaseasonswiththeanalysisof171samplessubmittedforviralrespiratorydisease.Increasedsensitivitywasfoundinoverallviraldetectionwiththissystemcomp...
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Real-TimeQuantitativePCRforDeterminingtheBurdenofPlasmodiumfalciparumParasitesduringPregnancyandInfancy
...obi,KenyaDepartmentofVeteran‘sAffairsMedicalCenter,Cleveland,OhioABSTRACTReal-timequantitativePCR(RTQ-PCR)providesaquick,accurate,andreproduciblequantificationofparasites.However,thevalueofRTQ-PCRforpredictingclinicaloutcomesofmalariaisunknown.Here,wecomparedRTQ-PCRtomicroscopyofbloodsmears,nested...
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Alteredexpressionprofileoftransportersintheinnermedullarycollectingductofaquaporin-1knockoutmice
...ionoftheseproteinswereconfirmedbyimmunocytochemistry.MessengerRNAprofiling(real-timeRT-PCR)revealedchangesinUT-A1,-ENaC,-ENaC,andAQP3transcriptabundancethatparalleledthechangesinproteinabundance.Thus,fromtheperspectiveoftransportproteins,theIMCDsofAQP1(/)micehaveasignificantlyalteredphenotype.Toaddr...
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血清HBV-DNA与HBV免疫学标志物之间的相关性研究
...志物之间的相关性。方法采用双盲实验，血清HBV-DNA采用Real-timePCR进行定量检测，血清免疫标志物采用ELISA法。结果在344份血清标本中，HBV五项免疫学标志物出现11种组合，主要有以下5种，组合Ⅰ为HBsAg（+）、HBeAg（+）、HBcAb（+）...
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AutomatedExtractionofViral-PathogenRNAandDNAforHigh-ThroughputQuantitativeReal-TimePCR
...is)asafront-endextractionsystemforhigh-throughput“in-house“quantitativereal-timePCRassayswasanalyzedandcomparedtothatofmanualextractionofplasmaandserumsamples(hepatitisCvirus[HCV]andhepatitisBvirus[HBV])andEDTA-bloodsamples(cytomegalovirus[CMV]andEpstein-Barrvirus[EBV]).Linearityofextractionwast...
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ComparisonofReal-TimePCRProtocolsforDifferentialLaboratoryDiagnosisofAmebiasis
...ctedwithEntamoebahistolytica,thespeciesthatcausesclinicalamebiasis.Distinctreal-timePCRprotocolshaverecentlybeenpublishedforidentificationofE.histolyticaanddifferentiationfromthemorphologicallyidenticalnonpathogenicEntamoebadispar.Inthisstudy,wecomparedthreeE.histolyticareal-timePCRtechniquespublish...
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开源qPCR：每个人都可以做DNA诊断
qPCR：英文全名是Real-timeQuantitativePCRDetectingSystem，即实时荧光定量核酸扩增检测系统，也叫实时定量基因扩增荧光检测系统，其中PCR指聚合酶链式反应。实时PCR热循环仪（Real-TimePCR）是一种功能强大的技术，可以检测大肠杆菌和...
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反复冻融对荧光实时定量PCR质粒标准品的影响研究
...准品ResearchonstabilityofplasmidDNAstandardsforfluorescencequantificationreal-timePCR WANGHeng-li,CHENHong-tao,KONGMei,etal.DepartmentofClinicalLaboratory,TheFifthAffiliatedHospitalofSunYat-senUniversity,Guangdong519000,China 【Abstract】ObjectiveToevaluatefreezingandthawingrepeatedlyi...
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QuantitativeReal-TimePCRAssayforDetectionofEhrlichiachaffeensis
...dification24April2003/Accepted7May2003 ABSTRACTTopAbstractTextReferencesAreal-timePCRassaywasdevelopedforthedetectionofEhrlichiachaffeensis.Theassayisspeciesspecificandprovidesquantitativeresultsintherange10to1010genecopies.Theassayisnotinhibitedbythepresenceoftick,human,ormouseDNAandiscompatiblew...
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基因与干细胞研究助推糖尿病治疗
...糖在尾静脉注射MafA蛋白质24小时时降至正常水平。RT-PCR、Real-time荧光定量PCR实验结果表明，MafA能够诱导小肠组织胰岛素基因转录增加。组织免疫荧光结果显示注射MafA蛋白质后小肠绒毛间的上皮细胞中存在有胰岛素阳性细胞。他...
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信号放大时间分辨荧光分析技术的研究进展及应用
...间分辨荧光分析；镧系元素螯合物 Studiesandapplicationsoftime-resolvedfluorescenceassaybasedonsignalamplification SONGNa-ling,ZHAOQi-ren. TheKeyLaboratoryofMolecularNuclearMedicine,InstituteofRadiationMedicine,ChineseAcademyofMedicalSciencesandPekingUnionMedicalCollege,Ti...
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Species-LevelIdentificationofStaphylococcalIsolatesbyReal-TimePCRandMeltCurveAnalysis
...ndNorthwesternUniversityFeinbergSchoolofMedicine,Evanston,IllinoisABSTRACTAreal-timePCRassaywasdevelopedtoidentifycommonstaphylococcalspecies.Asinglesetofconsensusprimerswasdesignedtoamplifyaportionofthe16SrRNAgene,andapairoffluorescenceresonanceenergytransferprobeswasusedtoidentifyspeciesbasedonthe...
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ComparisonofMethodsforDetectionofVacciniaVirusinPatientSpecimens
...rvices,Bethesda,Maryland20892ABSTRACTWeanalyzedashellvialcultureassay(SVA),real-timePCR,andadirectfluorescentantibodyassay(DFA)forrapiddetectionofvacciniavirusfromvaccinationsitesofDryvaxvaccinerecipients.Of47samplesassayed,100%werepositivebyPCR,89%werepositivebySVA,and40%werepositivebyDFA.DFAwaslim...
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临床常见腹泻致病菌的快速鉴定方法研究进展
...u/ml,检测时间由2～4周减少到1天。 1.4实时荧光PCR技术(realtimefluorescencePCR) 该技术在PCR反应体系中加入荧光基团,利用荧光信号积累实时检测整个PCR进程,最后通过标准曲线对未知模板进行定量的方法,其不仅具有普通PCR的高...