Mutagenesis, Vol. 18, No. 1, 45-51,
January 2003
© 2003 UK Environmental Mutagen Society/Oxford University Press
Recommendations for conducting the in vivo alkaline Comet assay
1 Novartis Pharma AG, Nervous Systems PRIDE, WSJ106.2.42, CH-4002 Basel, Switzerland, 2 AstraZeneca R&D Södertälje, Safety Assessment, S-151 85 Södertälje, Sweden, 3 Inventures, 1 Trevelyan Square, Boar Lane, Leeds LS1 6AE, UK, 4 Bayer AG, Toxicology, PO Box 101709, D-42096 Wuppertal, Germany, 5 GlaxoSmithKline, Park Road, Ware SG12 0DP, UK, 6 Syngenta Central Toxicology Laboratory, Alderley Park, Macclesfield SK10 4TJ, UK, 7 Rowett Research Institute, Greenburn Road, Bucksburn AB21 9SB, UK, 8 Health and Safety Executive, Industrial Chemicals Unit, Stanley Precinct, Bootle L20 3QZ, UK, 9 Universitätsklinikum Ulm, Abteilung Humangenetik, Albert Einstein Allee 11, D-89081 Ulm, Germany, 10 Aventis Pharma, 13 Quai Jules Guesde, BP 14, F-94403 Vitry-sur-Seine, France and 11 Integrated Laboratory Systems Inc., PO Box 13501, Research Triangle Park, NC 27709, USA
The in vivo alkaline single cell gel electrophoresis assay, hereafter the Comet assay, can be used to investigate the genotoxicity of industrial chemicals, biocides, agrochemicals and pharmaceuticals. The major advantages of this assay include the relative ease of application to any tissue of interest, the detection of multiple classes of DNA damage and the generation of data at the level of the single cell. These features give the Comet assay potential advantages over other in vivo test methods, which are limited largely to proliferating cells and/or a single tissue. The Comet assay has demonstrated its reliability in many testing circumstances and is, in general, considered to be acceptable for regulatory purposes. However, despite the considerable data published on the in vivo Comet assay and the general agreement within the international scientific community over many protocol-related issues, it was felt that a document giving detailed practical guidance on the protocol required for regulatory acceptance of the assay was required. In a recent meeting held in conjunction with the 4th International Comet Assay Workshop (Ulm, Germany, 22–25 July 2001) an expert panel reviewed existing data and recent developments of the Comet assay with a view to developing such a document. This paper is intended to act as an update to the more general guidelines which were published as a result of the International Workshop on Genotoxicity Test Procedures. The recommendations are also seen as a major step towards gaining more formal regulatory acceptance of the Comet assay.
12 To whom correspondence should be addressed. Tel: +41 61 32 45619; Fax: +41 61 32 49843; Email: andreas.hartmann{at}pharma.novartis.com
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