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Reproducibility of telomere length assessment: an international collaborative study.


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Authors

Martin-Ruiz, Carmen M 
Baird, Duncan 
Roger, Laureline 
Boukamp, Petra 
Krunic, Damir 

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.

Description

Keywords

Ageing, biomarker, human, telomeres, variation, Aging, Biomarkers, Blotting, Southern, DNA, Humans, International Cooperation, Real-Time Polymerase Chain Reaction, Reproducibility of Results, Telomere

Journal Title

Int J Epidemiol

Conference Name

Journal ISSN

0300-5771
1464-3685

Volume Title

Publisher

Oxford University Press (OUP)
Sponsorship
Isaac Newton Trust (1307 (k))
Cancer Research UK (16565)
The work was funded by the UK Medical Research Council [grant numbers G0601333 and G0500997 to T.vZ. and MC_UU_12019/1 to R.Co. and A.W.]; the New Dynamics of Ageing Initiative [grant number RES-353-25-0001 to R.Co.]; the Swedish Cancer Society [grant number 12 06249 to G.R. and U.S.]; the Swedish Research Council [grant number 90341301 to G.R. and U.S.]; the European Community's Seventh Framework Program FP7/2007-2011 [grant number 200950 to G.R. and U.S.]; the British Heart foundation [to V.C. and N.J.S.]; the NIHR Newcastle Biomedical Research Centre in Ageing and Chronic Disease [to C.M.M.R.]; the BMBF GerontoSys Stromal Aging [grant number 0315576A to P.B.]; UVA Konsortium [grant number 03NUK003A to P.B.]; the University of Utah Research Account [to R.Ca.]; the Association for International Cancer Research [grant number 10-0021 to D.B.]; and the Cunningham Trust [to P.S.].