Detection of DNA adducts by bioluminescence

Luminescent assay for detection ATP is very sensitive with limitation of 10^-17 moles. ATP Using styrene oxide as a model carcinogen we currently apply a luminescence technique to detect the very low levles of carcinogen-DNA adducts in vitro and in vivo. The bioluminescent assay of DNA adducts entails three consecutive steps: digestion of modified DNA to adducted dinucleoside monophsphate(*NpN) and normal nucleotide are hydrolyzed to nucleosides(N) by nuclease P1 and prostatic acid phosphomonesterase(PAP); incorporation of γ -P into *NpN from ATP catalyzed by polynucleotide kinase, while no incorporation of γ -P of ATP into normal nucleoside(N); detection of consumption of ATP by luminescence. This assay does not require separate manipulation because of the selective property of nuclease P1. One fmol of carcinogen-DNA adducts was detected by luminescent assay. A good correlation between results of luminescent assy and ³²P-postlabeling procedures has been observed. We detect 1 adduct in 10⁸ nucleotides for 20 μg DNA sample. The procedures of luminescent method is very simple and low-cost. It appears applicable to the ultrasensitive detection of low levels of DNA adducts without radioactive isotope.