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Reproducibility of telomere length assessment : an international collaborative study

Carmen M Martin-Ruiz, Duncan Baird, Laureline Roger, Petra Boukamp, Damir Krunic, Richard Cawthon, Martin M Dokter, Pim van der Harst, Sofie Bekaert (UGent) , Tim De Meyer (UGent) , et al.
(2015) INTERNATIONAL JOURNAL OF EPIDEMIOLOGY. 44(5). p.1673-1683
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Abstract
Background: Telomere length is a putative biomarker of ageing, morbidity and mortality. Its application is hampered by lack of widely applicable reference ranges and uncertainty regarding the present limits of measurement reproducibility within and between laboratories. Methods: We instigated an international collaborative study of telomere length assessment: 10 different laboratories, employing 3 different techniques [Southern blotting, single telomere length analysis (STELA) and real-time quantitative PCR (qPCR)] performed two rounds of fully blinded measurements on 10 human DNA samples per round to enable unbiased assessment of intra- and inter-batch variation between laboratories and techniques. Results: Absolute results from different laboratories differed widely and could thus not be compared directly, but rankings of relative telomere lengths were highly correlated (correlation coefficients of 0.63-0.99). Intra-technique correlations were similar for Southern blotting and qPCR and were stronger than inter-technique ones. However, inter-laboratory coefficients of variation (CVs) averaged about 10% for Southern blotting and STELA and more than 20% for qPCR. This difference was compensated for by a higher dynamic range for the qPCR method as shown by equal variance after z-scoring. Technical variation per laboratory, measured as median of intra- and inter-batch CVs, ranged from 1.4% to 9.5%, with differences between laboratories only marginally significant (P = 0.06). Gel-based and PCR-based techniques were not different in accuracy. Conclusions: Intra- and inter-laboratory technical variation severely limits the usefulness of data pooling and excludes sharing of reference ranges between laboratories. We propose to establish a common set of physical telomere length standards to improve comparability of telomere length estimates between laboratories.
Keywords
Ageing, EPIDEMIOLOGY, telomeres, variation, biomarker, human, CARDIOVASCULAR-DISEASE, SOUTHERN BLOTS, FLOW-CYTOMETRY, HUMAN-CELLS, MORTALITY, FIBROBLASTS, ASSOCIATION, BIOMARKER, DEMENTIA

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MLA
Martin-Ruiz, Carmen M., et al. “Reproducibility of Telomere Length Assessment : An International Collaborative Study.” INTERNATIONAL JOURNAL OF EPIDEMIOLOGY, vol. 44, no. 5, 2015, pp. 1673–83, doi:10.1093/ije/dyu191.
APA
Martin-Ruiz, C. M., Baird, D., Roger, L., Boukamp, P., Krunic, D., Cawthon, R., … von Zglinicki, T. (2015). Reproducibility of telomere length assessment : an international collaborative study. INTERNATIONAL JOURNAL OF EPIDEMIOLOGY, 44(5), 1673–1683. https://doi.org/10.1093/ije/dyu191
Chicago author-date
Martin-Ruiz, Carmen M, Duncan Baird, Laureline Roger, Petra Boukamp, Damir Krunic, Richard Cawthon, Martin M Dokter, et al. 2015. “Reproducibility of Telomere Length Assessment : An International Collaborative Study.” INTERNATIONAL JOURNAL OF EPIDEMIOLOGY 44 (5): 1673–83. https://doi.org/10.1093/ije/dyu191.
Chicago author-date (all authors)
Martin-Ruiz, Carmen M, Duncan Baird, Laureline Roger, Petra Boukamp, Damir Krunic, Richard Cawthon, Martin M Dokter, Pim van der Harst, Sofie Bekaert, Tim De Meyer, Goran Roos, Ulrika Svenson, Veryan Codd, Nilesh J Samani, Liane McGlynn, Paul G Shiels, Karen A Pooley, Alison M Dunning, Rachel Cooper, Andrew Wong, Andrew Kingston, and Thomas von Zglinicki. 2015. “Reproducibility of Telomere Length Assessment : An International Collaborative Study.” INTERNATIONAL JOURNAL OF EPIDEMIOLOGY 44 (5): 1673–1683. doi:10.1093/ije/dyu191.
Vancouver
1.
Martin-Ruiz CM, Baird D, Roger L, Boukamp P, Krunic D, Cawthon R, et al. Reproducibility of telomere length assessment : an international collaborative study. INTERNATIONAL JOURNAL OF EPIDEMIOLOGY. 2015;44(5):1673–83.
IEEE
[1]
C. M. Martin-Ruiz et al., “Reproducibility of telomere length assessment : an international collaborative study,” INTERNATIONAL JOURNAL OF EPIDEMIOLOGY, vol. 44, no. 5, pp. 1673–1683, 2015.
@article{7099072,
  abstract     = {{Background: Telomere length is a putative biomarker of ageing, morbidity and mortality. Its application is hampered by lack of widely applicable reference ranges and uncertainty regarding the present limits of measurement reproducibility within and between laboratories. 
Methods: We instigated an international collaborative study of telomere length assessment: 10 different laboratories, employing 3 different techniques [Southern blotting, single telomere length analysis (STELA) and real-time quantitative PCR (qPCR)] performed two rounds of fully blinded measurements on 10 human DNA samples per round to enable unbiased assessment of intra- and inter-batch variation between laboratories and techniques. 
Results: Absolute results from different laboratories differed widely and could thus not be compared directly, but rankings of relative telomere lengths were highly correlated (correlation coefficients of 0.63-0.99). Intra-technique correlations were similar for Southern blotting and qPCR and were stronger than inter-technique ones. However, inter-laboratory coefficients of variation (CVs) averaged about 10% for Southern blotting and STELA and more than 20% for qPCR. This difference was compensated for by a higher dynamic range for the qPCR method as shown by equal variance after z-scoring. Technical variation per laboratory, measured as median of intra- and inter-batch CVs, ranged from 1.4% to 9.5%, with differences between laboratories only marginally significant (P = 0.06). Gel-based and PCR-based techniques were not different in accuracy. 
Conclusions: Intra- and inter-laboratory technical variation severely limits the usefulness of data pooling and excludes sharing of reference ranges between laboratories. We propose to establish a common set of physical telomere length standards to improve comparability of telomere length estimates between laboratories.}},
  author       = {{Martin-Ruiz, Carmen M and Baird, Duncan and Roger, Laureline and Boukamp, Petra and Krunic, Damir and Cawthon, Richard and Dokter, Martin M and van der Harst, Pim and Bekaert, Sofie and De Meyer, Tim and Roos, Goran and Svenson, Ulrika and Codd, Veryan and Samani, Nilesh J and McGlynn, Liane and Shiels, Paul G and Pooley, Karen A and Dunning, Alison M and Cooper, Rachel and Wong, Andrew and Kingston, Andrew and von Zglinicki, Thomas}},
  issn         = {{0300-5771}},
  journal      = {{INTERNATIONAL JOURNAL OF EPIDEMIOLOGY}},
  keywords     = {{Ageing,EPIDEMIOLOGY,telomeres,variation,biomarker,human,CARDIOVASCULAR-DISEASE,SOUTHERN BLOTS,FLOW-CYTOMETRY,HUMAN-CELLS,MORTALITY,FIBROBLASTS,ASSOCIATION,BIOMARKER,DEMENTIA}},
  language     = {{eng}},
  number       = {{5}},
  pages        = {{1673--1683}},
  title        = {{Reproducibility of telomere length assessment : an international collaborative study}},
  url          = {{http://doi.org/10.1093/ije/dyu191}},
  volume       = {{44}},
  year         = {{2015}},
}
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