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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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治療性血管新生實驗研究的若干進展
...對照心肌ABCDExpressionchangesofbFGFR1afterratacutemyocardialinfarction(Real-timeRT-PCR,n=5)*#*#*#*#*#*#****//GEArrayTMQseriesMouseAngiogenesisGeneArray小鼠血管新生基因芯片(SuperArray,U.S.A.)非梗死區邊緣區心肌梗死大鼠心肌細胞內血管新生基因表達的變化(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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發現長期施肥對稻田土壤固碳自養菌多樣性影響
...期定位試驗爲平臺,採用PCR-克隆測序和實時熒光定量(Real-time)PCR技術,研究不施肥(CK),氮磷鉀肥(NPK)和秸稈還田(NPKS)3種長期施肥制度對稻田土壤固碳自養菌羣落結構及數量的影響。 通過分析固碳細菌cbbL基因文庫...
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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在白血病發病過程中的作用研究
...單鹼基錨定OligdT引物進行反轉錄,然後用特異引物進行realtimePCR擴增,從而檢測部分miRNAs在白血病細胞株K562中的表達,並與正常人骨髓表達情況相對比。結果通過realtimePCR分析,癌基因miR21在K562細胞中的表達量約爲(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與HBV免疫學標誌物之間的相關性研究
...志物之間的相關性。方法採用雙盲實驗,血清HBV-DNA採用Real-timePCR進行定量檢測,血清免疫標誌物採用ELISA法。結果在344份血清標本中,HBV五項免疫學標誌物出現11種組合,主要有以下5種,組合Ⅰ爲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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開源qPCR:每個人都可以做DNA診斷
qPCR:英文全名是Real-timeQuantitativePCRDetectingSystem,即實時熒光定量核酸擴增檢測系統,也叫實時定量基因擴增熒光檢測系統,其中PCR指聚合酶鏈式反應。實時PCR熱循環儀(Real-TimePCR)是一種功能強大的技術,可以檢測大腸桿菌和...
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反覆凍融對熒光實時定量PCR質粒標準品的影響研究
...準品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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基因與幹細胞研究助推糖尿病治療
...糖在尾靜脈注射MafA蛋白質24小時時降至正常水平。RT-PCR、Real-time熒光定量PCR實驗結果表明,MafA能夠誘導小腸組織胰島素基因轉錄增加。組織免疫熒光結果顯示注射MafA蛋白質後小腸絨毛間的上皮細胞中存在有胰島素陽性細胞。他...
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信號放大時間分辨熒光分析技術的研究進展及應用
...間分辨熒光分析;鑭系元素螯合物 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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臨牀常見腹瀉致病菌的快速鑑定方法研究進展
...u/ml,檢測時間由2~4周減少到1天。 1.4實時熒光PCR技術(realtimefluorescencePCR) 該技術在PCR反應體系中加入熒光基團,利用熒光信號積累實時檢測整個PCR進程,最後通過標準曲線對未知模板進行定量的方法,其不僅具有普通PCR的高...