Reliability and short-term intra-individual variability of telomere length measurement using monochrome multiplexing quantitative PCR.

<h4>Background</h4>Studies examining the association between telomere length and cancer risk have often relied on measurement of telomere length from a single blood draw using a real-time PCR technique. We examined the reliability of telomere length measurement using sequential samples c...

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Main Authors: Sangmi Kim, Dale P Sandler, Gleta Carswell, Clarice R Weinberg, Jack A Taylor
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2011-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0025774&type=printable
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author Sangmi Kim
Dale P Sandler
Gleta Carswell
Clarice R Weinberg
Jack A Taylor
author_facet Sangmi Kim
Dale P Sandler
Gleta Carswell
Clarice R Weinberg
Jack A Taylor
author_sort Sangmi Kim
collection DOAJ
description <h4>Background</h4>Studies examining the association between telomere length and cancer risk have often relied on measurement of telomere length from a single blood draw using a real-time PCR technique. We examined the reliability of telomere length measurement using sequential samples collected over a 9-month period.<h4>Methods and findings</h4>Relative telomere length in peripheral blood was estimated using a single tube monochrome multiplex quantitative PCR assay in blood DNA samples from 27 non-pregnant adult women (aged 35 to 74 years) collected in 7 visits over a 9-month period. A linear mixed model was used to estimate the components of variance for telomere length measurements attributed to variation among women and variation between time points within women. Mean telomere length measurement at any single visit was not significantly different from the average of 7 visits. Plates had a significant systematic influence on telomere length measurements, although measurements between different plates were highly correlated. After controlling for plate effects, 64% of the remaining variance was estimated to be accounted for by variance due to subject. Variance explained by time of visit within a subject was minor, contributing 5% of the remaining variance.<h4>Conclusion</h4>Our data demonstrate good short-term reliability of telomere length measurement using blood from a single draw. However, the existence of technical variability, particularly plate effects, reinforces the need for technical replicates and balancing of case and control samples across plates.
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spelling doaj-art-58838331c61644f689a43ff97e7184ae2025-08-20T02:34:48ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-0169e2577410.1371/journal.pone.0025774Reliability and short-term intra-individual variability of telomere length measurement using monochrome multiplexing quantitative PCR.Sangmi KimDale P SandlerGleta CarswellClarice R WeinbergJack A Taylor<h4>Background</h4>Studies examining the association between telomere length and cancer risk have often relied on measurement of telomere length from a single blood draw using a real-time PCR technique. We examined the reliability of telomere length measurement using sequential samples collected over a 9-month period.<h4>Methods and findings</h4>Relative telomere length in peripheral blood was estimated using a single tube monochrome multiplex quantitative PCR assay in blood DNA samples from 27 non-pregnant adult women (aged 35 to 74 years) collected in 7 visits over a 9-month period. A linear mixed model was used to estimate the components of variance for telomere length measurements attributed to variation among women and variation between time points within women. Mean telomere length measurement at any single visit was not significantly different from the average of 7 visits. Plates had a significant systematic influence on telomere length measurements, although measurements between different plates were highly correlated. After controlling for plate effects, 64% of the remaining variance was estimated to be accounted for by variance due to subject. Variance explained by time of visit within a subject was minor, contributing 5% of the remaining variance.<h4>Conclusion</h4>Our data demonstrate good short-term reliability of telomere length measurement using blood from a single draw. However, the existence of technical variability, particularly plate effects, reinforces the need for technical replicates and balancing of case and control samples across plates.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0025774&type=printable
spellingShingle Sangmi Kim
Dale P Sandler
Gleta Carswell
Clarice R Weinberg
Jack A Taylor
Reliability and short-term intra-individual variability of telomere length measurement using monochrome multiplexing quantitative PCR.
PLoS ONE
title Reliability and short-term intra-individual variability of telomere length measurement using monochrome multiplexing quantitative PCR.
title_full Reliability and short-term intra-individual variability of telomere length measurement using monochrome multiplexing quantitative PCR.
title_fullStr Reliability and short-term intra-individual variability of telomere length measurement using monochrome multiplexing quantitative PCR.
title_full_unstemmed Reliability and short-term intra-individual variability of telomere length measurement using monochrome multiplexing quantitative PCR.
title_short Reliability and short-term intra-individual variability of telomere length measurement using monochrome multiplexing quantitative PCR.
title_sort reliability and short term intra individual variability of telomere length measurement using monochrome multiplexing quantitative pcr
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0025774&type=printable
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AT gletacarswell reliabilityandshorttermintraindividualvariabilityoftelomerelengthmeasurementusingmonochromemultiplexingquantitativepcr
AT claricerweinberg reliabilityandshorttermintraindividualvariabilityoftelomerelengthmeasurementusingmonochromemultiplexingquantitativepcr
AT jackataylor reliabilityandshorttermintraindividualvariabilityoftelomerelengthmeasurementusingmonochromemultiplexingquantitativepcr