Reproducibility of telomere length assessment: an international collaborative study

Martin-Ruiz, C. M. et al. (2014) Reproducibility of telomere length assessment: an international collaborative study. International Journal of Epidemiology, 44(5), pp. 1673-1683. (doi:10.1093/ije/dyu191) (PMID:25239152) (PMCID:PMC4681105)

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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.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Shiels, Professor Paul and McGlynn, Dr Liane
Authors: Martin-Ruiz, C. M., Baird, D., Roger, L., Boukamp, P., Krunic, D., Cawthon, R., Dokter, M. M., van der Harst, P., Bekaert, S., de Meyer, T., Roos, G., Svenson, U., Codd, V., Samani, N. J., McGlynn, L., Shiels, P., Pooley, K. A., Dunning, A. M., Cooper, R., Wong, A., Kingston, A., and von Zglinicki, T.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cancer Sciences
Journal Name:International Journal of Epidemiology
Publisher:Oxford University Press
ISSN:0300-5771
ISSN (Online):1464-3685
Copyright Holders:Copyright © 2014 The Authors
First Published:First published in International Journal of Epidemiology 44(5):1673-1683
Publisher Policy:Reproduced under a Creative Commons License

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