A fluorescence-based screening assay for DNA damage induced by genotoxic industrial chemicals.
A rapid screening assay to detect chemically-induced DNA damage resulting from exposure of surrogate DNA to genotoxic compounds is reported. This assay is based on changes in the melting and annealing behavior observed for damaged DNA. Exposure of calf thymus DNA to genotoxic industrial chemicals reduced the extent to which the DNA annealed as measured using a double strand DNA selective fluorescent indicator dye. Formaldehyde, acrolein, crotonaldehyde and bromoethane showed the most prominent effects, chloroacetone and allylamine exhibited lesser effects, and acryrlonitrile showed no statistically significant assay response. The assay response for formaldehyde and crotonaldehyde were measured over the concentration range of 10-100 mM and 50-300 mM, respectively. This assay showed little response for the cytotoxic compounds phenol, cyclohexane and toluene but was sensitive to the effects of DNA damaging compounds such as mitomycin C and glutaraldehyde.
Kailasam
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US EPA National Exposure Research Laboratory-LV, Environmental Protection Agency, 944 E. Harmon Ave., Las Vegas, NV 89119, USA.
Rogers
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D000096:Acetone / Q000031:analogs & derivatives / Q000633:toxicity; D000171:Acrolein / Q000633:toxicity; D000447:Aldehydes / Q000633:toxicity; D000499:Allylamine / Q000633:toxicity; D004247:DNA / Q000737:chemistry* / Q000235:genetics; D004249:DNA Damage; D005453:Fluorescence; D005557:Formaldehyde / Q000633:toxicity; D006842:Hydrocarbons, Brominated / Q000633:toxicity; D009153:Mutagens / Q000633:toxicity*; D044366:Transition Temperature / Q000187:drug effects
D000447:Aldehydes; D006842:Hydrocarbons, Brominated; D009153:Mutagens; D000096:Acetone; D005557:Formaldehyde; D000499:Allylamine; C006973:chloroacetone; D000171:Acrolein; D004247:DNA; C427203:calf thymus DNA; C012796:2-butenal; C037311:bromoethane