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Statistical issues in the use of the comet assay

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Lovell, D. P., Omori, T.·2008·Mutagenesis

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Abstract

The comet or single-cell gel electrophoresis assay is now widely used in regulatory, mechanistic and biomonitoring studies using a range of in vitro and in vivo systems. Each of these has issues associated with the experimental design which determine to a large extent the statistical analyses than can be used. A key concept is that the experimental unit is the smallest 'amount' of experimental material that can be randomly assigned to a treatment: the animal for in vivo studies and the culture for in vitro studies. Biomonitoring studies, being observational rather than experimental, are vulnerable to confounding and biases. Critical factors in any statistical analysis include the identification of suitable end points, the choice of measure to represent the distribution of the comet end point in a sample of cells, estimates of variability between experimental units and the identification of the size of effects that could be considered biologically important. Power and sample size calculations can be used in conjunction with this information to identify optimum experimental sizes and provide help in combining the results of statistical analyses with other information to aid interpretation. Interpretation based upon the size of effects and their confidence intervals is preferred to that based solely upon statistical significance tests. Statistical issues associated with the design and subsequent analyses of current validation studies for the comet assay include the identification of acceptable levels of intra- and inter-laboratory repeatability and reproducibility and criteria for dichotomizing results into positive or negative.

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“The characteristics of the data, such as the experimental unit, the data type, and the (assumed) distribution, will guide various experimental features, including different levels of replication, number of test concentrations, and experimental repetitions (Lovell and Omori, 2008).”
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Authors: Lovell, D. P., & Omori, T.
Affiliations: University of Surrey (GB)
Kyoto University (JP)
Corresponding author: Lovell, D. P.
ORCID: Omori, T.
Venue: Mutagenesis
Publisher: Oxford University Press (OUP)
Published: 2008-02-17
Volume: 23
Issue: 3
Pages: 171-182
ISSN: 0267-8357 (ISSN-L)
1464-3804
DOI: 10.1093/mutage/gen015
OpenAlex: W2142519394
Cited by: 297
References: 48
Citations by year: 2026: 5 · 2025: 8 · 2024: 14 · 2023: 6 · 2022: 3 · 2021: 16 · 2020: 17 · 2019: 23 · 2018: 17 · 2017: 15 · 2016: 24 · 2015: 15 · 2014: 25 · 2013: 29 · 2012: 29
FWCI: 1.21
Type: journal article
Language: en
Open access: hybrid
doi.org
Licence: public-domain
Locations: Mutagenesis (publishedVersion, public-domain)
PubMed (publishedVersion)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Carcinogens and Genotoxicity Assessment
Health Sciences › Veterinary › Small Animals › Animal testing and alternatives
Social Sciences › Decision Sciences › Management Science and Operations Research › Optimal Experimental Design Methods
Keywords: comet assay, statistical design, biomonitoring, confounding and bias, sample size and power, effect size, confidence intervals, assay validation, repeatability and reproducibility, experimental units, mutagenicity testing
MeSH terms: Data Interpretation, Statistical, DNA Damage, Humans, Reproducibility of Results, Models, Statistical, Comet Assay

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