medlitГигиена и санитарияHygiene and Sanitation0016-99002412-0650Federal Scientific Center of Hygiene named after F.F. Erisman10.47470/0016-9900-2025-104-10-1369-1374qjfruxmedlit-5237Research ArticleМЕТОДЫ ГИГИЕНИЧЕСКИХ ИССЛЕДОВАНИЙMETHODS OF HYGIENIC AND EXPERIMENTAL INVESTIGATIONSВозможности применения ДНК-кометного анализа (обзор литературы)Possibilities of application of DNA Comet Assay (literature review)https://orcid.org/0000-0003-0028-2522ГоренскаяОльга ВладимировнаGorenskayaOlga V.

Канд. биол. наук, доцент, ст. науч. сотр. отд. генетической токсикологии ФБУН «ФНЦГ им. Ф.Ф. Эрисмана» Роспотребнадзора, 141014, Мытищи, Россия

e-mail: gorenskaya.ov@fncg.ru

PhD (Biology), docent, senior researcher, Department of genetic toxicology, Institute of Hygiene, Toxicology of Pesticides and Chemical Safety, Federal Scientific Center of Hygiene named after F.F. Erisman, Mytishchi, 141014, Russian Federation

e-mail: gorenskaya.ov@fncg.ru

gorenskaya.ov@fncg.ruhttps://orcid.org/0000-0002-5356-2600ДемидоваЮлия ВячеславовнаDemidovaYuliya V.

Мл. науч. сотр. отд. генетической токсикологии ФБУН «ФНЦГ им. Ф.Ф. Эрисмана» Роспотребнадзора, 141014, Мытищи, Россия

e-mail: demidova.yu@fncg.ru

Junior research, Department of genetic toxicology, Institute of Hygiene, Toxicology of Pesticides and Chemical Safety, Federal Scientific Center of Hygiene named after F.F. Erisman, Mytishchi, 141014, Russian Federation

e-mail: demidova.yu@fncg.ru

demidova.yu@fncg.ruhttps://orcid.org/0000-0002-4333-9288КотноваАлина ПетровнаKotnovaAlina P.

Канд. биол. наук, ст. науч. сотр. отд. генетической токсикологии ФБУН «ФНЦГ им. Ф.Ф. Эрисмана» Роспотребнадзора, 141014, Мытищи, Россия

e-mail: kotnova.ap@fncg.ru

PhD (Biology), senior researcher, Department of genetic toxicology, Institute of Hygiene, Toxicology of Pesticides and Chemical Safety of the Federal Scientific Center of Hygiene named after F.F. Erisman, Mytishchi, 141014, Russian Federation

e-mail: kotnova.ap@fncg.ru

kotnova.ap@fncg.ruhttps://orcid.org/0000-0001-9122-9465ИлюшинаНаталия АлексеевнаIlyushinaNatalia A.

Доктор биол. наук, зав. отд. генетической токсикологии ФБУН «ФНЦГ им. Ф.Ф. Эрисмана» Роспотребнадзора, 141014, Мытищи, Россия

e-mail: ilushina.na@fncg.ru

DSc (Biology), head, Department of genetic toxicology, Institute of Hygiene, Toxicology of Pesticides and Chemical Safety, Federal Scientific Center of Hygiene named after F.F. Erisman, Mytishchi, 141014, Russian Federation

e-mail: ilyushina.na@fncg.ru

ilushina.na@fncg.ruФБУН «Федеральный научный центр гигиены имени Ф.Ф. Эрисмана» РоспотребнадзораРоссияFederal Scientific Center of Hygiene named after F.F. ErismanRussian Federation2025171120251041013691374Copyright © Горенская О.В., Демидова Ю.В., Котнова А.П., Илюшина Н.А., 20252025Горенская О.В., Демидова Ю.В., Котнова А.П., Илюшина Н.А.Gorenskaya O.V., Demidova Y.V., Kotnova A.P., Ilyushina N.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.rjhas.ru/jour/article/view/5237

Для обеспечения санитарно-эпидемиологического благополучия населения необходим мониторинг среды обитания. В настоящее время существует множество методов определения неблагоприятных факторов, способных влиять на здоровье человека, в том числе нарушать генетические структуры в клетках. Одним из ключевых инструментов в генотоксикологии, который позволяет оценить повреждения ДНК на уровне отдельных клеток, является метод ДНК-кометного анализа. С момента появления в 1980-х годах он претерпел значительные модификации, направленные на повышение чувствительности, специфичности и универсальности. В данном обзоре рассмотрены принципы метода и основные протоколы, модификации классического кометного анализа, способы улучшения детекции разных типов повреждений ДНК и оценки репарационных процессов. Выполнен поиск релевантных исследований, опубликованных в реферативных базах данных Scopus, Web of Science, PubMed, РИНЦ, за период 1984–2024 гг. В обзор включена информация 47 полнотекстовых источников.

Заключение

Заключение. Метод ДНК-комет становится востребованным инструментом в исследованиях, направленных на обеспечение санитарно-эпидемиологического благополучия, выявление опасных для генетического здоровья населения факторов окружающей и производственной среды, оценку индивидуальной чувствительности клеток человека к генотоксикантам, с целью формирования групп риска и разработки мер профилактики. Новые методики с применением ферментов репарации для специфических типов повреждений и флуоресцентных красителей позволяют оценить процессы восстановления ДНК и типы повреждений. Использование криоконсервации цельной крови и лимфоцитов для последующего ДНК-анализа даёт возможность проводить крупномасштабные долгосрочные эпидемиологические исследования. Автоматизация анализа комет с помощью методов глубокого обучения повысит точность идентификации типов повреждений ДНК, что необходимо для выяснения механизмов действия конкретных веществ.

Участие авторов

Участие авторов: Горенская О.В. – сбор материала и обработка данных, написание текста, редактирование; Демидова Ю.В. – сбор материала; Котнова А.П. – сбор материала; Илюшина Н.А. – дизайн исследования, сбор материала и обработка данных, написание текста, редактирование. Все соавторы – утверждение окончательного варианта статьи, ответственность за целостность всех её частей.

Конфликт интересов

Конфликт интересов. Авторы декларируют отсутствие явных и потенциальных конфликтов интересов в связи с публикацией данной статьи.

Финансирование

Финансирование. Исследование выполнено в рамках государственного задания № 121090800086-7.

Поступила

Поступила: 30.06.2025 / Принята к печати: 15.10.2025 / Опубликована: 14.11.2025

To ensure the sanitary and epidemiological well-being of the population, it is necessary to monitor the state of the environment. Today, there is a wide range of methods available for identifying factors that can adversly affect human health, including those capable of disrupting genetic structures in cells. One of the key tools in genotoxicology that enables the assessment of DNA damage at the level of individual cells is the comet assay. Since its introduction in the 1980s, this method has undergone significant modifications aimed at increasing its sensitivity, specificity, and versatility of application. This review outlines the principles and basic protocols of the method, discusses modifications of the classical comet assay; methods for improving detection capability of different types of DNA damage and assessment of repair processes. A search for relevant studies published in the databases Scopus, Web of Science, PubMed, Russian Research Citation Index, for the period 1984–2024 was performed. The review includes information from forty seven full-text sources.

Conclusion

Conclusion. The DNA comet assay is becoming an increasingly in-demand tool in research aimed at ensuring the sanitary and epidemiological well-being of the population, identifying environmental and industrial factors dangerous to the genetic health of the population, assessing the individual sensitivity of human cells to genotoxicants to determine risk groups and implement preventive measures. The development of new protocols involving DNA repair enzymes specific to particular types of damage, along with the use of fluorescent dyes, allows evaluating DNA damage and repair processes. The use of cryopreservation of whole blood and lymphocytes for subsequent DNA analysis enables large-scale, long-term epidemiological studies. Automating comet assay analysis using deep learning methods will improve the accuracy of identifying DNA damage types, which is essential for clarifying the mechanisms of action of specific substances.

Contribution

Contribution: Gorenskaya O.V. – data collection and analysis, writing and editing the text; Demidova Yu.V. – data collection; Kotnova A.P. – data collection; Ilyushina N.A. – the study concept and design, data collection and analysis, writing and editing the text. All authors are responsible for the integrity of all parts of the manuscript and approval of the manuscript final version

Conflict of interest

Conflict of interest. The authors declare no conflict of interest.

Funding

Funding. The study was carried out within the framework of the state task No. 121090800086-7.

Received

Received: June 30, 2025 / Accepted: October 15, 2025 / Published: November 14, 2025

ДНК-кометный анализмодификациигенотоксичностьобзорComet AssaymodificationsgenotoxicantsreviewReferencesДурнев А.Д. Жанатаев А.К., Еремина Н.В. Генетическая токсикология. М.: Миттель Пресс; 2022. https://elibrary.ru/fbwcbtDurnev A.D. Zhanataev A.K., Eremina N.V. Genetic Toxicology [Geneticheskaya toksikologiya]. Moscow: Mittel Press; 2022. https://elibrary.ru/fbwcbt (in Russian)OECD Guidelines for the testing of chemicals. Test No. 486: Unscheduled DNA Synthesis (UDS) Test with Mammalian Liver Cells in Vivo; 1997. Available at: https://oecd.org/content/dam/oecd/en/publications/reports/1997/07/test-no-486-unscheduled-dna-synthesis-uds-test-with-mammalian-liver-cells-in-vivo_g1gh2977/9789264071520-en.pdfOECD Guidelines for the testing of chemicals. Test No. 486: Unscheduled DNA Synthesis (UDS) Test with Mammalian Liver Cells in Vivo; 1997. Available at: https://oecd.org/content/dam/oecd/en/publications/reports/1997/07/test-no-486-unscheduled-dna-synthesis-uds-test-with-mammalian-liver-cells-in-vivo_g1gh2977/9789264071520-en.pdfOECD Guidelines for the testing of chemicals. Test No. 482: Genetic Toxicology: DNA Damage and Repair, Unscheduled DNA Synthesis in Mammalian Cells in vitro; 1986. Available at: https://oecd.org/en/topics/sub-issues/testing-of-chemicals/replaced-and-deleted-test-guidelines.htmlOECD Guidelines for the testing of chemicals. Test No. 482: Genetic Toxicology: DNA Damage and Repair, Unscheduled DNA Synthesis in Mammalian Cells in vitro; 1986. Available at: https://oecd.org/en/topics/sub-issues/testing-of-chemicals/replaced-and-deleted-test-guidelines.htmlAzqueta A., Collins A.R. The essential comet assay: a comprehensive guide to measuring DNA damage and repair. Arch. Toxicol. 2013; 87(6): 949–68. https://doi.org/10.1007/s00204-013-1070-0Azqueta A., Collins A.R. The essential comet assay: a comprehensive guide to measuring DNA damage and repair. Arch. Toxicol. 2013; 87(6): 949–68. https://doi.org/10.1007/s00204-013-1070-0Rajendran M.S., Prabhakaran R., Vignesh S., Nagarathinam B. Comet assay: is it a sensitive tool in ecogenotoxicology? In: Gothandam K.M., Srinivasan R., Ranjan S., Dasgupta N., Lichtfouse E., eds. Environmental Biotechnology Volume 4. Environmental Chemistry for a Sustainable World, vol. 68. Cham: Springer; 2021: 181–99. https://doi.org/10.1007/978-3-030-77795-1_6Rajendran M.S., Prabhakaran R., Vignesh S., Nagarathinam B. Comet assay: is it a sensitive tool in ecogenotoxicology? In: Gothandam K.M., Srinivasan R., Ranjan S., Dasgupta N., Lichtfouse E., eds. Environmental Biotechnology Volume 4. Environmental Chemistry for a Sustainable World, vol. 68. Cham: Springer; 2021: 181–99. https://doi.org/10.1007/978-3-030-77795-1_6Milić M., Ceppi M., Bruzzone M., Azqueta A., Brunborg G., Godschalk R., et al. The hCOMET project: International database comparison of results with the comet assay in human biomonitoring. Baseline frequency of DNA damage and effect of main confounders. Mutat. Res. Rev. Mutat. Res. 2021; 787: 108371. https://doi.org/10.1016/j.mrrev.2021.108371Milić M., Ceppi M., Bruzzone M., Azqueta A., Brunborg G., Godschalk R., et al. The hCOMET project: International database comparison of results with the comet assay in human biomonitoring. Baseline frequency of DNA damage and effect of main confounders. Mutat. Res. Rev. Mutat. Res. 2021; 787: 108371. https://doi.org/10.1016/j.mrrev.2021.108371Collins A., Koppen G., Valdiglesias V., Dusinska M., Kruszewski M., Møller P., et al. The comet assay as a tool for human biomonitoring studies: the ComNet project. Mutat. Res. Rev. Mutat. Res. 2014; 759: 27–39. https://doi.org/10.1016/j.mrrev.2013.10.001Collins A., Koppen G., Valdiglesias V., Dusinska M., Kruszewski M., Møller P., et al. The comet assay as a tool for human biomonitoring studies: the ComNet project. Mutat. Res. Rev. Mutat. Res. 2014; 759: 27–39. https://doi.org/10.1016/j.mrrev.2013.10.001Collins A., Møller P., Gajski G., Vodenková S., Abdulwahed A., Anderson D., et al. Measuring DNA modifications with the comet assay: a compendium of protocols. Nat. Protoc. 2023; 18(3): 929–89. https://doi.org/10.1038/s41596-022-00754-yCollins A., Møller P., Gajski G., Vodenková S., Abdulwahed A., Anderson D., et al. Measuring DNA modifications with the comet assay: a compendium of protocols. Nat. Protoc. 2023; 18(3): 929–89. https://doi.org/10.1038/s41596-022-00754-yMøller P., Loft S. Oxidative damage to DNA and lipids as biomarkers of exposure to air pollution. Environ. Health Perspect. 2010; 118(8): 1126–36. https://doi.org/10.1289/ehp.0901725Møller P., Loft S. Oxidative damage to DNA and lipids as biomarkers of exposure to air pollution. Environ. Health Perspect. 2010; 118(8): 1126–36. https://doi.org/10.1289/ehp.0901725Жанатаев А.К., Дурнев А.Д., Середенин С.Б. Метод ДНК-комет в генотоксикологических исследованиях. Гигиена и санитария. 2011; 90(5): 86–90. https://elibrary.ru/ohikppZhanatayev A.K., Durnev A.D., Seredenin S.B. DNA comet assay in genotoxicological studies. Gigiena i Sanitaria (Hygiene and Sanitation, Russian journal). 2011; 90(5): 86–90. https://elibrary.ru/ohikpp (in Russian)Ostling O., Johanson K.J. Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. Biochem. Biophys. Res. Commun. 1984; 123(1): 291–8. https://doi.org/10.1016/0006-291x(84)90411-xOstling O., Johanson K.J. Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. Biochem. Biophys. Res. Commun. 1984; 123(1): 291–8. https://doi.org/10.1016/0006-291x(84)90411-xSingh N.P., McCoy M.T., Tice R.R., Schneider E.L. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp. Cell Res. 1988; 175(1): 184–91. https://doi.org/10.1016/0014-4827(88)90265-0Singh N.P., McCoy M.T., Tice R.R., Schneider E.L. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp. Cell Res. 1988; 175(1): 184–91. https://doi.org/10.1016/0014-4827(88)90265-0Bohn M., Heermann D.W. Diffusion-driven looping provides a consistent framework for chromatin organization. PLoS One. 2010; 5(8): e12218. https://doi.org/10.1371/journal.pone.0012218Bohn M., Heermann D.W. Diffusion-driven looping provides a consistent framework for chromatin organization. PLoS One. 2010; 5(8): e12218. https://doi.org/10.1371/journal.pone.0012218Dittmer T.A., Misteli T. The lamin protein family. Genome Biol. 2011; 12(5): 222. https://doi.org/10.1186/gb-2011-12-5-222Dittmer T.A., Misteli T. The lamin protein family. Genome Biol. 2011; 12(5): 222. https://doi.org/10.1186/gb-2011-12-5-222Tice R.R., Agurell E., Anderson D., Burlinson B., Hartmann A., Kobayashi H., et al. Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ. Mol. Mutagen. 2000; 35(3): 206–21. https://vk.cc/cR4KWdTice R.R., Agurell E., Anderson D., Burlinson B., Hartmann A., Kobayashi H., et al. Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ. Mol. Mutagen. 2000; 35(3): 206–21. https://vk.cc/cR4KWdCollins A.R. The comet assay for DNA damage and repair: principles, applications, and limitations. Mol. Biotechnol. 2004; 26(3): 249–61. https://doi.org/10.1385/MB:26:3:249Collins A.R. The comet assay for DNA damage and repair: principles, applications, and limitations. Mol. Biotechnol. 2004; 26(3): 249–61. https://doi.org/10.1385/MB:26:3:249Kumaravel T.S., Jha A.N. Reliable Comet assay measurements for detecting DNA damage induced by ionising radiation and chemicals. Mutat. Res. 2006; 605(1–2): 7–16. https://doi.org/10.1016/j.mrgentox.2006.03.002Kumaravel T.S., Jha A.N. Reliable Comet assay measurements for detecting DNA damage induced by ionising radiation and chemicals. Mutat. Res. 2006; 605(1–2): 7–16. https://doi.org/10.1016/j.mrgentox.2006.03.002Møller P., Azqueta A., Boutet-Robinet E., Koppen G., Bonassi S., Milić M., et al. Minimum Information for Reporting on the Comet Assay (MIRCA): recommendations for describing comet assay procedures and results. Nat. Protoc. 2020; 15(12): 3817–26. https://doi.org/10.1038/s41596-020-0398-1Møller P., Azqueta A., Boutet-Robinet E., Koppen G., Bonassi S., Milić M., et al. Minimum Information for Reporting on the Comet Assay (MIRCA): recommendations for describing comet assay procedures and results. Nat. Protoc. 2020; 15(12): 3817–26. https://doi.org/10.1038/s41596-020-0398-1Valverde M., Rojas E. Environmental and occupational biomonitoring using the Comet assay. Mutat. Res. 2009; 681(1): 93–109. https://doi.org/10.1016/j.mrrev.2008.11.001Valverde M., Rojas E. Environmental and occupational biomonitoring using the Comet assay. Mutat. Res. 2009; 681(1): 93–109. https://doi.org/10.1016/j.mrrev.2008.11.001Ladeira C., Koppen G., Scavone F., Giovannelli L. The comet assay for human biomonitoring: Effect of cryopreservation on DNA damage in different blood cell preparations. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2019; 843: 11–7. https://doi.org/10.1016/j.mrgentox.2019.02.002Ladeira C., Koppen G., Scavone F., Giovannelli L. The comet assay for human biomonitoring: Effect of cryopreservation on DNA damage in different blood cell preparations. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2019; 843: 11–7. https://doi.org/10.1016/j.mrgentox.2019.02.002Ladeira C., Møller P., Giovannelli L., Gajski G., Haveric A., Bankoglu E.E., et al. The comet assay as a tool in human biomonitoring studies of environmental and occupational exposure to chemicals – a systematic scoping review. Toxics. 2024; 12(4): 270. https://doi.org/10.3390/toxics12040270Ladeira C., Møller P., Giovannelli L., Gajski G., Haveric A., Bankoglu E.E., et al. The comet assay as a tool in human biomonitoring studies of environmental and occupational exposure to chemicals – a systematic scoping review. Toxics. 2024; 12(4): 270. https://doi.org/10.3390/toxics12040270Russo C., Acito M., Fatigoni C., Villarini M., Moretti M. B-comet assay (comet assay on buccal cells) for the evaluation of primary DNA damage in human biomonitoring studies. Int. J. Environ. Res. Public Health. 2020; 17(24): 9234. https://doi.org/10.3390/ijerph17249234Russo C., Acito M., Fatigoni C., Villarini M., Moretti M. B-comet assay (comet assay on buccal cells) for the evaluation of primary DNA damage in human biomonitoring studies. Int. J. Environ. Res. Public Health. 2020; 17(24): 9234. https://doi.org/10.3390/ijerph17249234Owiti N.A., Kaushal S., Martin L., Sly J., Swartz C.D., Fowler J., et al. Using the HepaCometChip assay for broad-spectrum DNA damage analysis. Curr. Protoc. 2022; 2(9): e563. https://doi.org/10.1002/cpz1.563Owiti N.A., Kaushal S., Martin L., Sly J., Swartz C.D., Fowler J., et al. Using the HepaCometChip assay for broad-spectrum DNA damage analysis. Curr. Protoc. 2022; 2(9): e563. https://doi.org/10.1002/cpz1.563Russo P., Lamonaca P., Milic M., Rojas E., Prinzi G., Cardaci V., et al. Biomarkers of DNA damage in COPD patients undergoing pulmonary rehabilitation: Integrating clinical parameters with genomic profiling. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2019; 843: 111–7. https://doi.org/10.1016/j.mrgentox.2019.04.003Russo P., Lamonaca P., Milic M., Rojas E., Prinzi G., Cardaci V., et al. Biomarkers of DNA damage in COPD patients undergoing pulmonary rehabilitation: Integrating clinical parameters with genomic profiling. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2019; 843: 111–7. https://doi.org/10.1016/j.mrgentox.2019.04.003Gajski G., Gerić M., Živković Semren T., Tariba Lovaković B., Oreščanin V., Pizent A. Application of the comet assay for the evaluation of DNA damage from frozen human whole blood samples: Implications for human biomonitoring. Toxicol. Lett. 2020; 319: 58–65. https://doi.org/10.1016/j.toxlet.2019.11.010Gajski G., Gerić M., Živković Semren T., Tariba Lovaković B., Oreščanin V., Pizent A. Application of the comet assay for the evaluation of DNA damage from frozen human whole blood samples: Implications for human biomonitoring. Toxicol. Lett. 2020; 319: 58–65. https://doi.org/10.1016/j.toxlet.2019.11.010Plitta-Michalak B.P., Ramos A., Stepien D., Trusiak M., Michalak M. The comet assay as a method for assessing DNA damage in cryopreserved samples. Cryo. Letters. 2024; 45(1): 1–15.Plitta-Michalak B.P., Ramos A., Stepien D., Trusiak M., Michalak M. The comet assay as a method for assessing DNA damage in cryopreserved samples. Cryo. Letters. 2024; 45(1): 1–15.Koppen G., De Prins S., Jacobs A., Nelen V., Schoeters G., Langie S.A.S. The comet assay in human biomonitoring: cryopreservation of whole blood and comparison with isolated mononuclear cells. Mutagenesis. 2018; 33(1): 41–7. https://doi.org/10.1093/mutage/gex034Koppen G., De Prins S., Jacobs A., Nelen V., Schoeters G., Langie S.A.S. The comet assay in human biomonitoring: cryopreservation of whole blood and comparison with isolated mononuclear cells. Mutagenesis. 2018; 33(1): 41–7. https://doi.org/10.1093/mutage/gex034Dalberto D., Nicolau C.C., Garcia A.L.H., Nordin A.P., Grivicich I., Silva J.D. Cytotoxic and genotoxic evaluation of cotinine using human neuroblastoma cells (SH-SY5Y). Genet. Mol. Biol. 2020; 43(2): e20190123. https://doi.org/10.1590/1678-4685-GMB-2019-0123Dalberto D., Nicolau C.C., Garcia A.L.H., Nordin A.P., Grivicich I., Silva J.D. Cytotoxic and genotoxic evaluation of cotinine using human neuroblastoma cells (SH-SY5Y). Genet. Mol. Biol. 2020; 43(2): e20190123. https://doi.org/10.1590/1678-4685-GMB-2019-0123Bonassi S., Ceppi M., Møller P., Azqueta A., Milić M., Neri M., et al. DNA damage in circulating leukocytes measured with the comet assay may predict the risk of death. Sci. Rep. 2021; 11(1): 16793. https://doi.org/10.1038/s41598-021-95976-7Bonassi S., Ceppi M., Møller P., Azqueta A., Milić M., Neri M., et al. DNA damage in circulating leukocytes measured with the comet assay may predict the risk of death. Sci. Rep. 2021; 11(1): 16793. https://doi.org/10.1038/s41598-021-95976-7Muruzabal D., Sanz-Serrano J., Sauvaigo S., Gützkow K.B., López de Cerain A., Vettorazzi A., et al. Novel approach for the detection of alkylated bases using the enzyme-modified comet assay. Toxicol. Lett. 2020; 330: 108–17. https://doi.org/10.1016/j.toxlet.2020.04.021Muruzabal D., Sanz-Serrano J., Sauvaigo S., Gützkow K.B., López de Cerain A., Vettorazzi A., et al. Novel approach for the detection of alkylated bases using the enzyme-modified comet assay. Toxicol. Lett. 2020; 330: 108–17. https://doi.org/10.1016/j.toxlet.2020.04.021Meng T., Zhang M., Song J., Dai Y., Duan H. Development of a co-culture model of mouse primary hepatocytes and splenocytes to evaluate xenobiotic genotoxicity using the medium-throughput Comet assay. Toxicol. In Vitro. 2020; 66: 104874. https://doi.org/10.1016/j.tiv.2020.104874Meng T., Zhang M., Song J., Dai Y., Duan H. Development of a co-culture model of mouse primary hepatocytes and splenocytes to evaluate xenobiotic genotoxicity using the medium-throughput Comet assay. Toxicol. In Vitro. 2020; 66: 104874. https://doi.org/10.1016/j.tiv.2020.104874Pellegri V., Gorbi G., Buschini A. DNA damage detection by Comet Assay on Daphnia magna: Application in freshwater biomonitoring. Sci. Total. Environ. 2020; 705: 135780. https://doi.org/10.1016/j.scitotenv.2019.135780Pellegri V., Gorbi G., Buschini A. DNA damage detection by Comet Assay on Daphnia magna: Application in freshwater biomonitoring. Sci. Total. Environ. 2020; 705: 135780. https://doi.org/10.1016/j.scitotenv.2019.135780Dusinska M., Collins A.R. Detection of oxidised purines and UV-induced photoproducts in DNA of single cells, by inclusion of lesion-specific enzymes in the comet assay. Altern. Lab. Anim. 1996; 24(3): 405–11.Dusinska M., Collins A.R. Detection of oxidised purines and UV-induced photoproducts in DNA of single cells, by inclusion of lesion-specific enzymes in the comet assay. Altern. Lab. Anim. 1996; 24(3): 405–11.Алексеева И.В., Бакман А.С., Яковлев Д.А., Кузнецов Н.А., Федорова О.С. Сравнительный анализ активности полиморфных вариантов урацил-ДНК-гликозилаз человека SMUG1 и MBD4. Молекулярная биология. 2021; 55(2): 277–88. https://doi.org/10.31857/S0026898421020026 https://elibrary.ru/ftnumqAlekseeva I.V., Bakman A.S., Iakovlev D.A., Kuznetsov N.A., Fedorova O.S. Comparative analysis of the activity of the polymorphic variants of human uracil-DNA-glycosylase SMUG1 and MBD4. Molekulyarnaya biologiya. 2021; 55(2): 277–88. https://doi.org/10.31857/S0026898421020026 https://elibrary.ru/ftnumq (in Russian)Catala G.N., Bestwick C.S., Russell W.R., Tortora K., Giovannelli L., Moyer M.P., et al. Folate, genomic stability and colon cancer: The use of single cell gel electrophoresis in assessing the impact of folate in vitro, in vivo and in human biomonitoring. Mutat. Res. Toxicol. Environ. Mutagen. 2019; 843: 73–80. https://doi.org/10.1016/j.mrgentox.2018.08.012Catala G.N., Bestwick C.S., Russell W.R., Tortora K., Giovannelli L., Moyer M.P., et al. Folate, genomic stability and colon cancer: The use of single cell gel electrophoresis in assessing the impact of folate in vitro, in vivo and in human biomonitoring. Mutat. Res. Toxicol. Environ. Mutagen. 2019; 843: 73–80. https://doi.org/10.1016/j.mrgentox.2018.08.012Holá M., Vágnerová R., Angelis K.J. Mutagenesis during plant responses to UVB radiation. Plant. Physiol. Biochem. 2015; 93: 29–33. https://doi.org/10.1016/j.plaphy.2014.12.013Holá M., Vágnerová R., Angelis K.J. Mutagenesis during plant responses to UVB radiation. Plant. Physiol. Biochem. 2015; 93: 29–33. https://doi.org/10.1016/j.plaphy.2014.12.013Илюшина Н.А., Егорова О.В., Котнова А.П., Горенская О.В., Аверьянова Н.С. Схема «Процедура проведения ДНК-кометного анализа для выявления модифицированных оснований и АР-сайтов в ДНК». Патент РФ 146952; 2025. https://elibrary.ru/ylwpbpIlyushina N.A., Egorova O.V., Kotnova A.P., Gorenskaya O.V., Averyanova N.S. Scheme "Procedure for conducting Comet Assay to identify modified bases and AP sites in DNA". Patent RF N 146952; 2025. https://elibrary.ru/ylwpbp (in Russian)Azqueta A., Langie S.A.S., Boutet-Robinet E., Duthie S., Ladeira C., Møller P., et al. DNA repair as a human biomonitoring tool: Comet assay approaches. Mutat. Res. Rev. Mutat. Res. 2019; 781: 71–87. https://doi.org/10.1016/j.mrrev.2019.03.002Azqueta A., Langie S.A.S., Boutet-Robinet E., Duthie S., Ladeira C., Møller P., et al. DNA repair as a human biomonitoring tool: Comet assay approaches. Mutat. Res. Rev. Mutat. Res. 2019; 781: 71–87. https://doi.org/10.1016/j.mrrev.2019.03.002Ngo L.P., Owiti N.A., Swartz C., Winters J., Su Y., Ge J., et al. Sensitive CometChip assay for screening potentially carcinogenic DNA adducts by trapping DNA repair intermediates. Nucleic Acids Res. 2020; 48: e13–e13. https://doi.org/10.1093/nar/gkz1077Ngo L.P., Owiti N.A., Swartz C., Winters J., Su Y., Ge J., et al. Sensitive CometChip assay for screening potentially carcinogenic DNA adducts by trapping DNA repair intermediates. Nucleic Acids Res. 2020; 48: e13–e13. https://doi.org/10.1093/nar/gkz1077Wentzel J.F., Gouws C., Huysamen C., Dyk Ev., Koekemoer G., Pretorius P.J. Assessing the DNA methylation status of single cells with the comet assay. Anal. Biochem. 2010; 400(2): 190–4. https://doi.org/10.1016/j.ab.2010.02.008Wentzel J.F., Gouws C., Huysamen C., Dyk Ev., Koekemoer G., Pretorius P.J. Assessing the DNA methylation status of single cells with the comet assay. Anal. Biochem. 2010; 400(2): 190–4. https://doi.org/10.1016/j.ab.2010.02.008Townsend T.A., Parrish M.C., Engelward B.P., Manjanatha M.G. The development and validation of EpiComet-Chip, a modified high-throughput comet assay for the assessment of DNA methylation status. Environ. Mol. Mutagen. 2017; 58: 508–21. https://doi.org/10.1002/em.22101Townsend T.A., Parrish M.C., Engelward B.P., Manjanatha M.G. The development and validation of EpiComet-Chip, a modified high-throughput comet assay for the assessment of DNA methylation status. Environ. Mol. Mutagen. 2017; 58: 508–21. https://doi.org/10.1002/em.22101Danson S., Ranson M., Denneny O., Cummings J., Ward T.H. Validation of the comet-X assay as a pharmacodynamic assay for measuring DNA cross-linking produced by the novel anticancer agent RH1 during a phase I clinical trial. Cancer Chemother. Pharmacol. 2007; 60(6): 851–61. https://doi.org/10.1007/s00280-007-0432-9Danson S., Ranson M., Denneny O., Cummings J., Ward T.H. Validation of the comet-X assay as a pharmacodynamic assay for measuring DNA cross-linking produced by the novel anticancer agent RH1 during a phase I clinical trial. Cancer Chemother. Pharmacol. 2007; 60(6): 851–61. https://doi.org/10.1007/s00280-007-0432-9Shaposhnikov S., Azqueta A., Henriksson S., Meier S., Gaivão I., Huskisson N.H., et al. Twelve-gel slide format optimised for comet assay and fluorescent in situ hybridisation. Toxicol. Lett. 2010; 195(1): 31–4. https://doi.org/10.1016/j.toxlet.2010.02.017Shaposhnikov S., Azqueta A., Henriksson S., Meier S., Gaivão I., Huskisson N.H., et al. Twelve-gel slide format optimised for comet assay and fluorescent in situ hybridisation. Toxicol. Lett. 2010; 195(1): 31–4. https://doi.org/10.1016/j.toxlet.2010.02.017Elje E., Hesler M., Rundén-Pran E., Mann P., Mariussen E., Wagner S., et al. The comet assay applied to HepG2 liver spheroids. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2019; 845: 403033. https://doi.org/10.1016/j.mrgentox.2019.03.006Elje E., Hesler M., Rundén-Pran E., Mann P., Mariussen E., Wagner S., et al. The comet assay applied to HepG2 liver spheroids. Mutat. Res. Genet. Toxicol. Environ. Mutagen. 2019; 845: 403033. https://doi.org/10.1016/j.mrgentox.2019.03.006Møller P., Knudsen L.E., Loft S., Wallin H. The comet assay as a rapid test in biomonitoring occupational exposure to DNA-damaging agents and effect of confounding factors. Cancer Epidemiol. Biomarkers Prev. 2000; 9(10): 1005–15.Møller P., Knudsen L.E., Loft S., Wallin H. The comet assay as a rapid test in biomonitoring occupational exposure to DNA-damaging agents and effect of confounding factors. Cancer Epidemiol. Biomarkers Prev. 2000; 9(10): 1005–15.Azqueta A., Ladeira C., Giovannelli L., Boutet-Robinet E., Bonassi S., Neri M., et al. Application of the comet assay in human biomonitoring: An hCOMET perspective. Mutat. Res. Rev. Mutat. Res. 2020; 783: 108288. https://doi.org/10.1016/j.mrrev.2019.108288Azqueta A., Ladeira C., Giovannelli L., Boutet-Robinet E., Bonassi S., Neri M., et al. Application of the comet assay in human biomonitoring: An hCOMET perspective. Mutat. Res. Rev. Mutat. Res. 2020; 783: 108288. https://doi.org/10.1016/j.mrrev.2019.108288Softić A., Merdović N., Dlakić V., Mrđanović E., Mahmutović L., Ler D., et al. Comet Assay in the Digital Era: A Review of the Use of Artificial Intelligence for the Analysis of DNA Damage Based on the Results of the Comet Assay. In: Jarm T., Šmerc R., Mahnič-Kalamiza S., eds. 9th European Medical and Biological Engineering Conference. IFMBE Proceedings, vol 112. Cham: Springer; 2024. https://doi.org/10.1007/978-3-031-61625-9_20Softić A., Merdović N., Dlakić V., Mrđanović E., Mahmutović L., Ler D., et al. Comet Assay in the Digital Era: A Review of the Use of Artificial Intelligence for the Analysis of DNA Damage Based on the Results of the Comet Assay. In: Jarm T., Šmerc R., Mahnič-Kalamiza S., eds. 9th European Medical and Biological Engineering Conference. IFMBE Proceedings, vol 112. Cham: Springer; 2024. https://doi.org/10.1007/978-3-031-61625-9_20

The authors declare that there are no conflicts of interest present.