Impartial comparative analysis of measurement of leukocyte telomere length/DNA content by Southern blots and qPCR
Open accessAbstract
Telomere length/DNA content has been measured in epidemiological/clinical settings with the goal of testing a host of hypotheses related to the biology of human aging, but often the conclusions of these studies have been inconsistent. These inconsistencies may stem from various reasons, including the use of different telomere length measurement techniques. Here, we report the first impartial evaluation of measurements of leukocyte telomere length by Southern blot of the terminal restriction fragments and quantitative PCR (qPCR) of telomere DNA content, expressed as the ratio of telomeric product (T)/single copy gene (S) product. Blind measurements on the same samples from 50 donors were performed in two independent laboratories on two different occasions. Both the qPCR and Southern blots displayed highly reproducible results as shown by r values > 0.9 for the correlations between results obtained by either method on two occasions. The inter-assay CV measurement for the qPCR was 6.45%, while that of the Southern blots was 1.74%. The relation between the results generated by Southern blots versus those generated by qPCR deviated from linearity. We discuss the ramifications of these findings with regard to measurements of telomere length/DNA content in epidemiological/clinical circumstances.
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Different results may derive from different DNA extraction and assay methods and different laboratories.
“(c) different results may derive from different DNA extraction and assay methods and different laboratories (Aubert et al., 2012; Aviv et al., 2011; Cunningham et al., 2013; Montpetit et al., 2014; Nieratschker et al., 2013);”
From Psychiatric disorders and leukocyte telomere length: Underlying mechanisms linking mental illness with cellular aging (2015) · cites this paper as Aviv et al., 2011
The qPCR method used to measure mean leukocyte telomere length was validated and shown to be significantly associated with the Southern blot method.
“This method was validated for mean telomere length measurement and shown to be significantly associated with the Southern blot method (51).”
From Determinants of telomere attrition over 1 year in healthy older women: stress and health behaviors matter (2015) · cites this paper as [51]
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- Authors
- Aviv, A., Hunt, S. C., Lin, J., Cao, X., Kimura, M., & Blackburn, E.
- Affiliations
- Rutgers New Jersey Medical School (US)
- Rutgers, The State University of New Jersey (US)
- Corresponding author
- Aviv, A.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2011-08-08
- Volume
- 39
- Issue
- 20
- Pages
- e134-e134
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- PMC
- PMC3203599
- OpenAlex
- W2124142246
- Cited by
- 343
- References
- 24
- Citations by year
- 2026: 8 · 2025: 9 · 2024: 7 · 2023: 9 · 2022: 17 · 2021: 19 · 2020: 19 · 2019: 23 · 2018: 32 · 2017: 35 · 2016: 34 · 2015: 34 · 2014: 39 · 2013: 32 · 2012: 22
- FWCI
- 20.94
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Nucleic Acids Research (publishedVersion, cc-by-nc)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Rutgers University Community Repository (Rutgers University) (submittedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- eScholarship (California Digital Library) (submittedVersion, cc-by-nc)
- Funders
- National Institutes of Health
- National Institute on Aging (AG030678, R01AG030678, AG18734, R01AG020132, AG20132, R01AG018734)
- Topics
- Health Sciences › Medicine › Physiology › Telomeres, Telomerase, and Senescence
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Aging › Genetics, Aging, and Longevity in Model Organisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
- Keywords
- leukocyte telomere length, Southern blot, real-time PCR, telomere length measurement, inter-method comparison, measurement reproducibility, aging
- MeSH terms
- Adult, Aged, DNA, Female, Humans, Leukocytes, Male, Middle Aged, Blotting, Southern, Reproducibility of Results, Polymerase Chain Reaction, Telomere, Telomere Homeostasis
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