Single cell gel assay

There are reports indicating that arsenic compounds can damage cellular DNA, thereby acting as genotoxins, as assessed by the comet or single-cell gel assay. Some reports indicate that arsenite possesses this activity (5-7), whereas another indicates that although the methylated trivalent species are genotoxic, arsenite itself is not (8). 

Meyers, C.D., Fairbairn. D.W, O Neill, K.L. (1993) Measuring the repair of HjOj-induced DNA single strand breaks using the single cell gel assay. Cytobios, 14, 147-153... 

Gasiorowski, K., Brocos, B. DNA repair of hydrogen peroxide-induced damage in human lymphocytes in the presence of four antimutagens. A study with alkaline single cell gel electrophoresis (comet assay). Cellular Molecular Biology Letters, Vol.6, (2001), pp. 897-911, ISSN 1425-8153... 

In Vivo (or In Vitro) Single-Cell Gel Electrophoresis Test for DNA Damage (Comet Assay)... 

Sardas S, Aygun N, Gamli M, et al Use of alkaline comet assay (single cell gel electrophoresis technique) to detect DNA damage in lymphocytes of operating room personnel occupationally exposed to anaesthetic gases. Mutat Res 418(2-3) 93-100, 1998... 

H., Miyamae, Y, Rojas, E. et al. (2000) Single cell gel/comet assay guidelines for in vitro and in vivo genetic toxicology testing. Environmental and Molecular Mutagenesis, 35, 206-221. 

NT044 Tsuda, S., N. Matsusaka, S. Ueno, N. Susa, and Y. F. Sasaki. The influence of antioxidants on cigarette smoke-induced DNA single-strand breaks in mouse organs a preliminary study with the alkaline single cell gel electrophoresis assay. Toxicol Sci 2000 54(1) 104-109. 

Martin, F.L., Cole, K.J., Orme, M.H., Grover, P.L., Phillips, D.H. Venitt, S. (1999) The DNA repair inhibitors hydroxyurea and cytosine arabinoside enhance the sensitivity of the alkaline single-cell gel electrophoresis ( comet ) assay in metabohcally-competent MCL-5 cells. Mutat Res., 445, 21-43... 

Lee, J.-G, Madden, M.C., Reed, W., Adler, K. Devlin, R. (1996) The use of the single cell gel electrophoresis assay in detecting DNA single strand breaks in lung cells in vitro. Toxicol, appl. Pharmacol., 141, 195-204... 

Yendle, J.E., Tinwell, H., Elliott, B.M. Ashby, J. (1997) The genetic toxicity of time importance of DNA-unwinding time to the outcome of single-cell gel electrophoresis assays. Mutat. Res., 375, 125-136... 

Sauvaigo S, Serres C, Signorini N, Emonet N, Richard M-J, Cadet J (1998) Use of the single-cell gel electrophoresis assay for the immonofluorescent detection of specific DNA damage. Anal Bio-chem 259 1-7... 

Ralph S, Petras M. 1998. Caged amphibian tadpoles and in situ genotoxicity monitoring of aquatic environments with the alkaline single cell gel electrophoresis (comet) assay. Mutat Res 413 235-250. 

This test, called the Comet Assay or single-cell gel-electrophoresis assay, allows the degree of DNA damage to be determined within a nucleic cell population. The principle of the method is based on the microelectrophoresis of nuclei of isolated cells, under basic conditions, on agarose gel (the whole being observed under a fluorescence microscope). 

Modifications of the standard battery may be necessary for some classes, e.g., antibiotics which are toxic to bacteria or e.g., for compounds like topoisomerase inhibitors which interfere with the mammalian cell replication system. A selection of additional assays is being proposed, further modifications may be acceptable via discussion in the ICH Maintenance Process. Alternative strategies may consider assays like the in vivo Comet assay (single cell gel electrophoresis measuring DNA strand breaks) or gene mutation tests with transgenic animals or in vivo DNA adduct studies. 

UHL M, HELMA c and KNASMULLER s (2000) Evaluation of the single cell gel electrophoresis assay with human hepatoma (Hep G2) cells, Mutat Res 468, 213-225. 

Measurement of Drug-Induced DNA Interstrand Crosslinking Using the Single-Cell Gel Electrophoresis (Comet) Assay... 

Hartmann, A. and Speit, G. (1994) Comparative investigations of the genotoxic effects of metals in the single cell gel (SCG) assay and the sister chromatid exchanges (SCE). Mutat. Res. 346,49-56. 

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