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The Comet Assay: Assessment of In Vitro and In Vivo DNA Damage

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Book cover Genotoxicity Assessment

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1044))

Abstract

Rapid industrialization and pursuance of a better life have led to an increase in the amount of chemicals in the environment, which are deleterious to human health. Pesticides, automobile exhausts, and new chemical entities all add to air pollution and have an adverse effect on all living organisms including humans. Sensitive test systems are thus required for accurate hazard identification and risk assessment. The Comet assay has been used widely as a simple, rapid, and sensitive tool for assessment of DNA damage in single cells from both in vitro and in vivo sources as well as in humans. Already, the in vivo comet assay has gained importance as the preferred test for assessing DNA damage in animals for some international regulatory guidelines. The advantages of the in vivo comet assay are its ability to detect DNA damage in any tissue, despite having non-proliferating cells, and its sensitivity to detect genotoxicity. The recommendations from the international workshops held for the comet assay have resulted in establishment of guidelines. The in vitro comet assay conducted in cultured cells and cell lines can be used for screening large number of compounds and at very low concentrations. The in vitro assay has also been automated to provide a high-throughput screening method for new chemical entities, as well as environmental samples. This chapter details the in vitro comet assay using the 96-well plate and in vivo comet assay in multiple organs of the mouse.

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Acknowledgements

The authors gratefully acknowledge the funding from CSIR, New Delhi, under its network projects (NWP34, NWP35) and OLP 009. The funding from UK India Education and Research Initiative (UKIERI) standard award to Institute of Life Sciences, Ahmedabad University, Ahmedabad, India (IND/CONT/E/11-12/217), and the University of Bradford is gratefully acknowledged. Funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement no. 263147 (NanoValid—Development of reference methods for hazard identification, risk assessment and LCA of engineered nanomaterials) is also acknowledged.

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Authors and Affiliations

  1. Institute of Life Sciences, School of Science and Technology, Ahmedabad University, Ahmedabad, Gujarat, India

    Mahima Bajpayee

  2. The Science Hub, Lucknow, UP, India

    Mahima Bajpayee

  3. Institute of Life Sciences, School of Science & Technology, Ahmedabad University, Ahmedabad, Gujarat, India

    Ashutosh Kumar & Alok Dhawan

  4. Nanomaterial Toxicology Group, Council of Scientific and Industrial Research (CSIR), Indian Institute of Toxicology Research, Lucknow, UP, India

    Alok Dhawan

Authors
  1. Mahima Bajpayee
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  2. Ashutosh Kumar
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  3. Alok Dhawan
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Editor information

Editors and Affiliations

  1. School of Science and Technology, Ahmedabad University Institute of Life Sciences, Ahmedabad, India

    Alok Dhawan

  2. School of Science and Technology, Ahmedabad University Institute of Life Sciences, Ahmedabad, India

    Mahima Bajpayee

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Bajpayee, M., Kumar, A., Dhawan, A. (2013). The Comet Assay: Assessment of In Vitro and In Vivo DNA Damage. In: Dhawan, A., Bajpayee, M. (eds) Genotoxicity Assessment. Methods in Molecular Biology, vol 1044. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-529-3_17

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  • DOI: https://doi.org/10.1007/978-1-62703-529-3_17

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-528-6

  • Online ISBN: 978-1-62703-529-3

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