Abstract
The comet assay is a widely used test for the detection of DNA damage and repair activity. However, there are interlaboratory differences in reported levels of baseline and induced damage in the same experimental systems. These differences may be attributed to protocol differences, although it is difficult to identify the relevant conditions because detailed comet assay procedures are not always published. Here, we present a Consensus Statement for the Minimum Information for Reporting Comet Assay (MIRCA) providing recommendations for describing comet assay conditions and results. These recommendations differentiate between ‘desirable’ and ‘essential’ information: ‘essential’ information refers to the precise details that are necessary to assess the quality of the experimental work, whereas ‘desirable’ information relates to technical issues that might be encountered when repeating the experiments. Adherence to MIRCA recommendations should ensure that comet assay results can be easily interpreted and independently verified by other researchers.
Highlights
The comet assay is a widely used test for the detection of DNA damage and repair activity
A recent review summarized the plenitude of procedure descriptions and technical recommendations for comet assays that have been published in the past 20 years, and highlighted the problem of inter-laboratory variation in DNA damage levels[15]
The main issue is that the comet assay does not directly measure the number of specific DNA lesions, but rather measures the migration of DNA in agarose gels as a result of the relaxation produced by strand breaks under alkaline conditions
Summary
The comet assay is a widely used test for the detection of DNA damage and repair activity. There are interlaboratory differences in reported levels of baseline and induced damage in the same experimental systems These differences may be attributed to protocol differences, it is difficult to identify the relevant conditions because detailed comet assay procedures are not always published. The OECD guideline on the in vivo comet assay (TG489), which was developed by multiple authors at various different institutions, is the most authoritative set of recommendations for the reporting of in vivo comet assay procedures and results[18] It does not cover in vitro experiments and biomonitoring studies, or endpoints other than DNA strand breaks, and the level of detail is limited in some aspects. For the use of the comet assay in molecular epidemiology studies, we recommend reading the Strengthening the Reporting of Observational Studies in Epidemiology–Molecular
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