METHOD OF ASSAY OF GENOTOXIC EFFECT OF RADIATION-CHEMICAL EFFECTS

摘要

FIELD: biological medicine, ecology. SUBSTANCE: blood is treated with mixture to obtain nucleoides and its background fluorescence is determined at λ_в = 350 nm and at λ_f = 450 nm, stained with 4',6-diamidino-2-phenylindole and fluorescence is determined again. Then DNA solution is added at concentration 5-10 mcg/ml and fluorescence is determined again and nucleoide DNA concentration is calculated per a 1 leukocyte in analyzed blood sample by the following formula: : L where ИФ_o - intensity of background nucleoide fluorescence; ИФ₁ - nucleoide fluorescence intensity after their staining; ИФ₂ - nucleoide fluorescence intensity after addition of DNA solution; K - coefficient of nucleoide and DNA dilution degree; L - leukocytes number/ml sample. Lysing mixture: solution containing 2.0 M sodium chloride, 0.1 M Trilon B, 0.01 M tris, 0.5% Triton X-100, pH 7.8-8.2. Genotoxicity of radiation-chemical effect is diagnosed if nucleoide DNA concentration is decreased by 30%, not less, as compared with that in intact organisms. EFFECT: improved method of assay, decreased analysis time (up to 5-10 min) and cost of analysis (by 10-20-fold). 2 cl, 1 tbl