Reliability and Short-Term Intra-Individual Variability of Telomere Length Measurement Using Monochrome Multiplexing Quantitative PCR 英文参考文献.docVIP

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Reliability and Short-Term Intra-Individual Variability of Telomere Length Measurement Using Monochrome Multiplexing Quantitative PCR 英文参考文献.doc

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Reliability and Short-Term Intra-Individual Variability of Telomere Length Measurement Using Monochrome Multiplexing Quantitative PCR 英文参考文献

ReliabilityandShort-TermIntra-IndividualVariabilityof TelomereLengthMeasurementUsingMonochrome MultiplexingQuantitativePCR SangmiKim1*,DaleP.Sandler1,GletaCarswell2,ClariceR.Weinberg3,JackA.Taylor1,2 1EpidemiologyBranch,NationalInstituteofEnvironmentalHealthSciences,ResearchTrianglePark,NorthCarolina,UnitedStatesofAmerica,2LaboratoryofMolecular Carcinogenesis; National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina, United States of America, 3Biostatistics Branch, National InstituteofEnvironmentalHealthSciences,ResearchTrianglePark,NorthCarolina,UnitedStatesofAmerica Abstract Background: Studies examining the association between telomere length and cancer risk have often relied on measurementoftelomerelengthfromasingleblooddrawusingareal-timePCRtechnique.Weexaminedthereliabilityof telomerelengthmeasurementusingsequentialsamplescollectedovera9-monthperiod. Methods and Findings: Relative telomere length in peripheral blood was estimated using a single tube monochrome multiplexquantitativePCRassayinbloodDNAsamplesfrom27non-pregnantadultwomen(aged35to74years)collected in 7 visits over a9-month period. A linear mixed model was used toestimate the components of variance for telomere length measurements attributed to variation among women and variation between time points within women. Mean telomerelengthmeasurementatanysinglevisitwasnotsignificantlydifferentfromtheaverageof7visits.Plateshada significantsystematicinfluenceontelomerelengthmeasurements,althoughmeasurementsbetweendifferentplateswere highlycorrelated.Aftercontrollingforplateeffects,64%oftheremainingvariancewasestimatedtobeaccountedforby varianceduetosubject.Varianceexplainedbytimeofvisitwithinasubjectwasminor,contributing5%oftheremaining variance. Conclusion:Ourdatademonstrategoodshort-termreliabilityoftelomerelengthmeasurementusingbloodfromasingle draw.However,theexistenceoftechnicalvariability,particularlyplateeffects,reinforcestheneedfortechnicalreplicates andbalancing