Detection of DNA adducts by bioluminescence

Shunqing Xu; Xianglin Tan; Qunfeng Yao; Min He; Yikai Zhou; Jian Chen

doi:10.1117/12.440151

14 September 2001 Detection of DNA adducts by bioluminescence

Shunqing Xu, Xianglin Tan, Qunfeng Yao, Min He, Yikai Zhou, Jian Chen

Proceedings Volume 4414, International Conference on Sensor Technology (ISTC 2001); (2001) https://doi.org/10.1117/12.440151
Event: International Conference on Sensing units and Sensor Technology, 2001, Wuhan, China

Abstract

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 levels 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 monophosphate and normal nucleotide are hydrolyzed to nucleosides (N) by nuclease P1 and prostatic acid phosphomonesterase (PAP); incorporation of (gamma) -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 assay and ³²P-postlabeling procedures has been observed. We detect 1 adduct in 10⁸ nucleotides for 10(mu) g DNA sample. The procedures of luminescent method is very simple and low- cost. IT appears applicable to the ultra sensitive detection of low levels of DNA adducts without radioactive isotope.

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Shunqing Xu, Xianglin Tan, Qunfeng Yao, Min He, Yikai Zhou, and Jian Chen "Detection of DNA adducts by bioluminescence", Proc. SPIE 4414, International Conference on Sensor Technology (ISTC 2001), (14 September 2001); https://doi.org/10.1117/12.440151

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