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The alkaline comet assay: towards validation in biomonitoring of DNA damaging exposures.
Basic Clin Pharmacol Toxicol. 2006 Apr; 98(4):336-45.BC

Abstract

Generation of DNA damage is considered to be an important initial event in carcinogenesis. The single cell gel electrophoresis (comet) assay is a technically simple and fast method that detects genotoxicity in virtually any mammalian cell type without requirement for cell culture. This review discusses the strength of the comet assay in biomonitoring at its present state of validation. The simple version of the alkaline comet assay detects DNA migration caused by strand breaks, alkaline labile sites, and transient repair sites. By incubation with bacterial glycosylase/endonuclease enzymes, broad classes of oxidative DNA damage, alkylations, and ultraviolet light-induced photoproducts are detected as additional DNA migration. The most widely measured enzyme sensitive sites have been those detected by formamidopyrimidine DNA glycosylase (FPG) and endonuclease III (ENDOIII). Reports from biomonitoring studies show that the basal level of DNA damage in leukocytes is influenced be a variety of lifestyle and environmental exposures, including exercise, air pollution, sunlight, and diet. Although not all types of carcinogenic exposures should be expected to damage DNA in leukocytes, the comet assay is a valuable method for detection of genotoxic exposure in humans. However, the predictive value of the comet assay is unknown because it has not been investigated in prospective cohort studies. Also, it is important that the performance of the assay is investigated in multi-laboratory validation trials. As a tool in risk assessment the comet assay can be used in characterization of hazards.

Authors+Show Affiliations

Institute of Public Health, University of Copenhagen, Øster Farimagsgade 5, Building B, 2nd Floor, P.O. Box 2099, DK-1014 Copenhagen K, Denmark. p.moller@pubhealth.ku.dk

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Review

Language

eng

PubMed ID

16623855

Citation

Møller, Peter. "The Alkaline Comet Assay: Towards Validation in Biomonitoring of DNA Damaging Exposures." Basic & Clinical Pharmacology & Toxicology, vol. 98, no. 4, 2006, pp. 336-45.
Møller P. The alkaline comet assay: towards validation in biomonitoring of DNA damaging exposures. Basic Clin Pharmacol Toxicol. 2006;98(4):336-45.
Møller, P. (2006). The alkaline comet assay: towards validation in biomonitoring of DNA damaging exposures. Basic & Clinical Pharmacology & Toxicology, 98(4), 336-45.
Møller P. The Alkaline Comet Assay: Towards Validation in Biomonitoring of DNA Damaging Exposures. Basic Clin Pharmacol Toxicol. 2006;98(4):336-45. PubMed PMID: 16623855.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The alkaline comet assay: towards validation in biomonitoring of DNA damaging exposures. A1 - Møller,Peter, PY - 2006/4/21/pubmed PY - 2006/10/27/medline PY - 2006/4/21/entrez SP - 336 EP - 45 JF - Basic & clinical pharmacology & toxicology JO - Basic Clin Pharmacol Toxicol VL - 98 IS - 4 N2 - Generation of DNA damage is considered to be an important initial event in carcinogenesis. The single cell gel electrophoresis (comet) assay is a technically simple and fast method that detects genotoxicity in virtually any mammalian cell type without requirement for cell culture. This review discusses the strength of the comet assay in biomonitoring at its present state of validation. The simple version of the alkaline comet assay detects DNA migration caused by strand breaks, alkaline labile sites, and transient repair sites. By incubation with bacterial glycosylase/endonuclease enzymes, broad classes of oxidative DNA damage, alkylations, and ultraviolet light-induced photoproducts are detected as additional DNA migration. The most widely measured enzyme sensitive sites have been those detected by formamidopyrimidine DNA glycosylase (FPG) and endonuclease III (ENDOIII). Reports from biomonitoring studies show that the basal level of DNA damage in leukocytes is influenced be a variety of lifestyle and environmental exposures, including exercise, air pollution, sunlight, and diet. Although not all types of carcinogenic exposures should be expected to damage DNA in leukocytes, the comet assay is a valuable method for detection of genotoxic exposure in humans. However, the predictive value of the comet assay is unknown because it has not been investigated in prospective cohort studies. Also, it is important that the performance of the assay is investigated in multi-laboratory validation trials. As a tool in risk assessment the comet assay can be used in characterization of hazards. SN - 1742-7835 UR - https://www.unboundmedicine.com/medline/citation/16623855/The_alkaline_comet_assay:_towards_validation_in_biomonitoring_of_DNA_damaging_exposures_ L2 - https://doi.org/10.1111/j.1742-7843.2006.pto_167.x DB - PRIME DP - Unbound Medicine ER -