{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T14:08:05Z","timestamp":1790777285704,"version":"4.1.0"},"reference-count":38,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2016,5,3]],"date-time":"2016-05-03T00:00:00Z","timestamp":1462233600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2016,5,3]],"date-time":"2016-05-03T00:00:00Z","timestamp":1462233600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Measurement of telomere length is widely used in epidemiologic studies. Insufficient standardization of the measurements processes has, however, complicated the comparison of results between studies. We aimed to investigate whether DNA extraction methods have an influence on measured values of relative telomere length (RTL) and whether this has consequences for epidemiological studies. We performed four experiments with RTL measurement in quadruplicate by qPCR using DNA extracted with different methods: 1) a standardized validation experiment including three extraction methods (magnetic-particle-method EZ1, salting-out-method INV, phenol-chloroform-isoamyl-alcohol PCI) each in the same 20 samples demonstrated pronounced differences in RTL with lowest values with EZ1 followed by INV and PCI-isolated DNA; 2) a comparison of 307 samples from an epidemiological study showing EZ1-measurements 40% lower than INV-measurements; 3) a matching-approach of two similar non-diseased control groups including 143 pairs of subjects revealed significantly shorter RTL in EZ1 than INV-extracted DNA (0.844\u2009\u00b1\u20090.157 vs. 1.357\u2009\u00b1\u20090.242); 4) an association analysis of RTL with prevalent cardiovascular disease detected a stronger association with INV than with EZ1-extracted DNA. In summary, DNA extraction methods have a pronounced influence on the measured RTL-values. This might result in spurious or lost associations in epidemiological studies under certain circumstances.<\/jats:p>","DOI":"10.1038\/srep25398","type":"journal-article","created":{"date-parts":[[2016,5,3]],"date-time":"2016-05-03T05:13:35Z","timestamp":1462252415000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":46,"title":["Influence of DNA extraction methods on relative telomere length measurements and its impact on epidemiological studies"],"prefix":"10.1038","volume":"6","author":[{"given":"Julia","family":"Raschenberger","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"Lamina","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Margot","family":"Haun","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Kollerits","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Coassin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Boes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ludmilla","family":"Kedenko","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"K\u00f6ttgen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Florian","family":"Kronenberg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2016,5,3]]},"reference":[{"key":"BFsrep25398_CR1","doi-asserted-by":"publisher","first-page":"569","DOI":"10.1038\/350569a0","volume":"350","author":"EH Blackburn","year":"1991","unstructured":"Blackburn, E. H. Structure and function of telomeres. Nature 350, 569\u2013573 (1991).","journal-title":"Nature"},{"key":"BFsrep25398_CR2","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1016\/0022-2836(78)90294-2","volume":"120","author":"EH Blackburn","year":"1978","unstructured":"Blackburn, E. H. & Gall, J. G. A tandemly repeated sequence at the termini of the extrachromosomal ribosomal RNA genes in Tetrahymena. J. Mol. Biol. 120, 33\u201353 (1978).","journal-title":"J. Mol. Biol."},{"key":"BFsrep25398_CR3","doi-asserted-by":"publisher","first-page":"405","DOI":"10.1016\/0092-8674(85)90170-9","volume":"43","author":"CW Greider","year":"1985","unstructured":"Greider, C. W. & Blackburn, E. H. Identification of a specific telomere terminal transferase activity in Tetrahymena extracts. Cell 43, 405\u2013413 (1985).","journal-title":"Cell"},{"key":"BFsrep25398_CR4","doi-asserted-by":"publisher","first-page":"532","DOI":"10.1038\/sj.onc.1205080","volume":"21","author":"T de Lange","year":"2002","unstructured":"de Lange, T. Protection of mammalian telomeres. Oncogene 21, 532\u2013540 (2002).","journal-title":"Oncogene"},{"key":"BFsrep25398_CR5","doi-asserted-by":"publisher","first-page":"1194","DOI":"10.1016\/j.cell.2013.05.039","volume":"153","author":"C Lopez-Otin","year":"2013","unstructured":"Lopez-Otin, C., Blasco, M. A., Partridge, L., Serrano, M. & Kroemer, G. The hallmarks of aging. Cell 153, 1194\u20131217 (2013).","journal-title":"Cell"},{"key":"BFsrep25398_CR6","doi-asserted-by":"publisher","first-page":"e43","DOI":"10.1126\/sageke.2004.51.pe43","volume":"2004","author":"A Aviv","year":"2004","unstructured":"Aviv, A. Telomeres and human aging: facts and fibs. Sci. Aging Knowledge. Environ. 2004, e43 (2004).","journal-title":"Sci. Aging Knowledge. Environ"},{"key":"BFsrep25398_CR7","doi-asserted-by":"publisher","first-page":"399","DOI":"10.1111\/joim.12083","volume":"274","author":"SE Bojesen","year":"2013","unstructured":"Bojesen, S. E. Telomeres and human health. J. Intern. Med. 274, 399\u2013413 (2013).","journal-title":"J. Intern. Med."},{"key":"BFsrep25398_CR8","doi-asserted-by":"publisher","first-page":"822","DOI":"10.1161\/ATVBAHA.111.237271","volume":"32","author":"M Weischer","year":"2012","unstructured":"Weischer, M. et al. Short telomere length, myocardial infarction, ischemic heart disease and early death. Arterioscler. Thromb. Vasc. Biol. 32, 822\u2013829 (2012).","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"BFsrep25398_CR9","doi-asserted-by":"publisher","first-page":"459","DOI":"10.1093\/jnci\/djt016","volume":"105","author":"M Weischer","year":"2013","unstructured":"Weischer, M. et al. Short telomere length, cancer survival and cancer risk in 47102 individuals. J. Natl. Cancer Inst. 105, 459\u2013468 (2013).","journal-title":"J. Natl. Cancer Inst."},{"key":"BFsrep25398_CR10","doi-asserted-by":"publisher","first-page":"274","DOI":"10.1038\/nrcardio.2013.30","volume":"10","author":"F Fyhrquist","year":"2013","unstructured":"Fyhrquist, F., Saijonmaa, O. & Strandberg, T. The roles of senescence and telomere shortening in cardiovascular disease. Nat. Rev. Cardiol. 10, 274\u2013283 (2013).","journal-title":"Nat. Rev. Cardiol"},{"key":"BFsrep25398_CR11","doi-asserted-by":"publisher","first-page":"1649","DOI":"10.1161\/ATVBAHA.110.205492","volume":"30","author":"P Willeit","year":"2010","unstructured":"Willeit, P. et al. Cellular aging reflected by leukocyte telomere length predicts advanced atherosclerosis and cardiovascular disease risk. Arterioscler. Thromb. Vasc. Biol. 30, 1649\u20131656 (2010).","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"BFsrep25398_CR12","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1001\/jama.2010.897","volume":"304","author":"P Willeit","year":"2010","unstructured":"Willeit, P. et al. Telomere length and risk of incident cancer and cancer mortality. JAMA 304, 69\u201375 (2010).","journal-title":"JAMA"},{"key":"BFsrep25398_CR13","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1016\/S0140-6736(07)60071-3","volume":"369","author":"SW Brouilette","year":"2007","unstructured":"Brouilette, S. W. et al. Telomere length, risk of coronary heart disease and statin treatment in the West of Scotland Primary Prevention Study: a nested case-control study. Lancet 369, 107\u2013114 (2007).","journal-title":"Lancet"},{"key":"BFsrep25398_CR14","doi-asserted-by":"publisher","first-page":"469","DOI":"10.1016\/j.atherosclerosis.2013.05.027","volume":"229","author":"J Raschenberger","year":"2013","unstructured":"Raschenberger, J. et al. The association of relative telomere length with symptomatic peripheral arterial disease: Results from the CAVASIC Study. Atherosclerosis 229, 469\u2013474 (2013).","journal-title":"Atherosclerosis"},{"key":"BFsrep25398_CR15","doi-asserted-by":"publisher","first-page":"11887","DOI":"10.1038\/srep11887","volume":"5","author":"J Raschenberger","year":"2015","unstructured":"Raschenberger, J. et al. Association of relative telomere length with progression of chronic kidney disease in two cohorts: effect modification by smoking and diabetes. Scientific Reports 5, 11887 (2015).","journal-title":"Scientific Reports"},{"key":"BFsrep25398_CR16","doi-asserted-by":"publisher","first-page":"e112483","DOI":"10.1371\/journal.pone.0112483","volume":"9","author":"P Willeit","year":"2014","unstructured":"Willeit, P. et al. Leucocyte Telomere Length and Risk of Type 2 Diabetes Mellitus: New Prospective Cohort Study and Literature-Based Meta-Analysis. PLos. ONE. 9, e112483 (2014).","journal-title":"PLos. ONE."},{"key":"BFsrep25398_CR17","doi-asserted-by":"publisher","first-page":"529","DOI":"10.1016\/j.atherosclerosis.2015.08.020","volume":"242","author":"J Raschenberger","year":"2015","unstructured":"Raschenberger, J. et al. Association of relative telomere length with cardiovascular disease in a large chronic kidney disease cohort: the GCKD study. Atherosclerosis 242, 529\u2013534 (2015).","journal-title":"Atherosclerosis"},{"key":"BFsrep25398_CR18","doi-asserted-by":"publisher","first-page":"e21485","DOI":"10.1371\/journal.pone.0021485","volume":"6","author":"U Svenson","year":"2011","unstructured":"Svenson, U. et al. Blood cell telomere length is a dynamic feature. PLos. ONE. 6, e21485 (2011).","journal-title":"PLos. ONE."},{"key":"BFsrep25398_CR19","doi-asserted-by":"publisher","first-page":"155","DOI":"10.1111\/joim.12149","volume":"275","author":"J Huzen","year":"2014","unstructured":"Huzen, J. et al. Telomere length loss due to smoking and metabolic traits. J. Intern. Med. 275, 155\u2013163 (2014).","journal-title":"J. Intern. Med."},{"key":"BFsrep25398_CR20","doi-asserted-by":"publisher","first-page":"e1004191","DOI":"10.1371\/journal.pgen.1004191","volume":"10","author":"M Weischer","year":"2014","unstructured":"Weischer, M., Bojesen, S. E. & Nordestgaard, B. G. Telomere shortening unrelated to smoking, body weight, physical activity and alcohol intake: 4,576 general population individuals with repeat measurements 10 years apart. PLos Genet. 10, e1004191 (2014).","journal-title":"PLos Genet."},{"key":"BFsrep25398_CR21","doi-asserted-by":"publisher","first-page":"458","DOI":"10.1038\/345458a0","volume":"345","author":"CB Harley","year":"1990","unstructured":"Harley, C. B., Futcher, A. B. & Greider, C. W. Telomeres shorten during ageing of human fibroblasts. Nature 345, 458\u2013460 (1990).","journal-title":"Nature"},{"key":"BFsrep25398_CR22","doi-asserted-by":"publisher","first-page":"1596","DOI":"10.1038\/nprot.2010.124","volume":"5","author":"M Kimura","year":"2010","unstructured":"Kimura, M. et al. Measurement of telomere length by the Southern blot analysis of terminal restriction fragment lengths. Nat. Protoc. 5, 1596\u20131607 (2010).","journal-title":"Nat. Protoc."},{"key":"BFsrep25398_CR23","doi-asserted-by":"publisher","first-page":"e47","DOI":"10.1093\/nar\/30.10.e47","volume":"30","author":"RM Cawthon","year":"2002","unstructured":"Cawthon, R. M. Telomere measurement by quantitative PCR. Nucleic Acids Res. 30, e47 (2002).","journal-title":"Nucleic Acids Res."},{"key":"BFsrep25398_CR24","doi-asserted-by":"publisher","first-page":"1673","DOI":"10.1093\/ije\/dyu191","volume":"44","author":"CM Martin-Ruiz","year":"2015","unstructured":"Martin-Ruiz, C. M. et al. Reproducibility of telomere length assessment: an international collaborative study. Int. J. Epidemiol. 44, 1673\u20131683 (2015).","journal-title":"Int. J. Epidemiol."},{"key":"BFsrep25398_CR25","doi-asserted-by":"publisher","first-page":"497","DOI":"10.1038\/hdy.2010.113","volume":"105","author":"T Horn","year":"2010","unstructured":"Horn, T., Robertson, B. C. & Gemmell, N. J. The use of telomere length in ecology and evolutionary biology. Heredity (Edinb.) 105, 497\u2013506 (2010).","journal-title":"Heredity (Edinb.)"},{"key":"BFsrep25398_CR26","doi-asserted-by":"publisher","first-page":"299","DOI":"10.1111\/2041-210X.12161","volume":"5","author":"DH Nussey","year":"2014","unstructured":"Nussey, D. H. et al. Measuring telomere length and telomere dynamics in evolutionary biology and ecology. Methods Ecol. Evol. 5, 299\u2013310 (2014).","journal-title":"Methods Ecol. Evol"},{"key":"BFsrep25398_CR27","doi-asserted-by":"publisher","first-page":"2047","DOI":"10.1158\/1055-9965.EPI-13-0409","volume":"22","author":"JM Cunningham","year":"2013","unstructured":"Cunningham, J. M. et al. Telomere length varies by DNA extraction method: implications for epidemiologic research. Cancer Epidemiol. Biomarkers Prev. 22, 2047\u20132054 (2013).","journal-title":"Cancer Epidemiol. Biomarkers Prev"},{"key":"BFsrep25398_CR28","doi-asserted-by":"publisher","first-page":"877","DOI":"10.1186\/1756-0500-7-877","volume":"7","author":"J Denham","year":"2014","unstructured":"Denham, J., Marques, F. Z. & Charchar, F. J. Leukocyte telomere length variation due to DNA extraction method. BMC. Res. Notes 7, 877 (2014).","journal-title":"BMC. Res. Notes"},{"key":"BFsrep25398_CR29","doi-asserted-by":"publisher","first-page":"851","DOI":"10.1373\/clinchem.2007.102046","volume":"54","author":"B Rantner","year":"2008","unstructured":"Rantner, B. et al. Association between the UGT1A1 TA-repeat polymorphism and bilirubin concentration in patients with intermittent claudication: results from the CAVASIC Study. Clin. Chem. 54, 851\u2013857 (2008).","journal-title":"Clin. Chem."},{"key":"BFsrep25398_CR30","doi-asserted-by":"publisher","first-page":"375","DOI":"10.2337\/diabetes.55.02.06.db05-0747","volume":"55","author":"IM Heid","year":"2006","unstructured":"Heid, I. M. et al. Genetic architecture of the APM1 gene and its influence on adiponectin plasma levels and parameters of the metabolic syndrome in 1,727 healthy Caucasians. Diabetes 55, 375\u2013384 (2006).","journal-title":"Diabetes"},{"key":"BFsrep25398_CR31","doi-asserted-by":"publisher","first-page":"611","DOI":"10.1373\/clinchem.2008.112797","volume":"55","author":"SA Bustin","year":"2009","unstructured":"Bustin, S. A. et al. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin. Chem. 55, 611\u2013622 (2009).","journal-title":"Clin. Chem."},{"key":"BFsrep25398_CR32","doi-asserted-by":"publisher","first-page":"e45","DOI":"10.1093\/nar\/29.9.e45","volume":"29","author":"MW Pfaffl","year":"2001","unstructured":"Pfaffl, M. W. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 29, e45 (2001).","journal-title":"Nucleic Acids Res."},{"key":"BFsrep25398_CR33","doi-asserted-by":"publisher","first-page":"e45","DOI":"10.1093\/nar\/gkp045","volume":"37","author":"JM Ruijter","year":"2009","unstructured":"Ruijter, J. M. et al. Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR data. Nucleic Acids Res. 37, e45 (2009).","journal-title":"Nucleic Acids Res."},{"key":"BFsrep25398_CR34","doi-asserted-by":"publisher","first-page":"307","DOI":"10.1016\/S0140-6736(86)90837-8","volume":"1","author":"JM Bland","year":"1986","unstructured":"Bland, J. M. & Altman, D. G. Statistical methods for assessing agreement between two methods of clinical measurement. Lancet 1, 307\u2013310 (1986).","journal-title":"Lancet"},{"key":"BFsrep25398_CR35","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1093\/clinchem\/46.1.100","volume":"46","author":"RF Martin","year":"2000","unstructured":"Martin, R. F. General deming regression for estimating systematic bias and its confidence interval in method-comparison studies. Clin. Chem. 46, 100\u2013104 (2000).","journal-title":"Clin. Chem."},{"key":"BFsrep25398_CR36","doi-asserted-by":"publisher","first-page":"1463","DOI":"10.1002\/sim.4780091210","volume":"9","author":"K Linnet","year":"1990","unstructured":"Linnet, K. Estimation of the linear relationship between the measurements of two methods with proportional errors. Stat. Med. 9, 1463\u20131473 (1990).","journal-title":"Stat. Med."},{"key":"BFsrep25398_CR37","doi-asserted-by":"crossref","unstructured":"Carroll, R. J., Ruppert, D. & Stefanski, L. A. Measurement Error in Nonlinear Models. Chapman & Hall\/CRC, (1995)","DOI":"10.1007\/978-1-4899-4477-1"},{"key":"BFsrep25398_CR38","doi-asserted-by":"publisher","first-page":"570","DOI":"10.1002\/ajhb.22690","volume":"27","author":"DT Eisenberg","year":"2015","unstructured":"Eisenberg, D. T., Kuzawa, C. W. & Hayes, M. G. Improving qPCR telomere length assays: Controlling for well position effects increases statistical power. Am. J. Hum. Biol. 27, 570\u2013575 (2015).","journal-title":"Am. J. Hum. Biol."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/srep25398","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep25398.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep25398.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,1,4]],"date-time":"2023-01-04T09:45:59Z","timestamp":1672825559000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/srep25398"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,5,3]]},"references-count":38,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2016,7,6]]}},"alternative-id":["BFsrep25398"],"URL":"https:\/\/doi.org\/10.1038\/srep25398","relation":{"has-review":[{"id-type":"doi","id":"10.3410\/f.726327765.793521663","asserted-by":"object"}]},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,5,3]]},"assertion":[{"value":"9 November 2015","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"13 April 2016","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"3 May 2016","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing financial interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"25398"}}