Methodological aspects of DNA-comet assay in vivo in pre-clinical research

Е.А. Gajdaj,  ORCID 0000-0002-5295-6384; 
A.A. Dorofeeva, ORCID0000-0002-8738-2296;
K.L. Kryshen, ORCID 0000-0003-1451-7716;
D.S.Gajdaj, ORCID 0000-0002-8773-5717

Institute of Pre-Clinical Research Ltd.
188663, Russia, Leningradskiy region, Vsevolozhskiy district, Kuzmolovskiy, Zavodskaya st., 3-245

Е-mail: gajdaj.ea@doclinika.ru

Abstract

Prolonged influence of adverse factors of environment on any biological system is accompanied by accumulation of damages in cell’s DNA and change of reparation systems activity that may lead to emergence of fixed mutations, oncogenesis and cell death.

Methodological and methodical bases of the Comet assay for detection of genomic DNA damages is presented in the review. During the last years, many analytical techniques have been developed to estimate DNA damage, however not all of them possess sufficient sensitivity and specificity for monitoring variety of DNA damages. The method, first described by Ostling and Johansson in 1984, possesses sensitivity necessary for registration of DNA damage and repair occurring on a single cell level and may be used for assessment of genome integrity. Improvements and modifications of the comet assay significantly increased its sensitivity and expanded the application area; however, they did not affect on the basic principles.

Currently, the comet assay is used in various fields: in studies of the genotoxic effect of chemicals (including pharmaceuticals), the study of DNA damage repair, apoptosis, clinical studies on prenatal diagnosis, susceptibility to cancer, cancer therapy, cataract. The comet assay is an integral part of biomonitoring programs: the influence of the diet on the body, environmental factors, changes in metabolism and physiological state, aging of the body; to study the mechanisms of radioprotective effects and the formation of the radio-adaptive response; research on ecology.

The comet assay has a number of features that affect the effectiveness of the method. Among these features are the possibility of freezing the material, temperature conditions for processing the material, light conditions, etc. To ensure repeatability of reliable results, it is necessary to unify the measures taken in the comet assay to avoid variability of results.

Full text avaliable in Russain only

Acknowledgements

The study was performed without external funding.

Authors’ contributions

Е.А. Gajdaj – data collection and analysis, study concept and design, writing and editing of the text

A.A. Dorofeeva – setting up an experiment, literary data collection, writing of the text

K.L. Kryshen, D.S. Gajdaj – literary data collection, editing of the text

Conflict of interest

The authors declare no conflict of interest.

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