Microplate enzyme-linked immunosorbent assay for the detection of primary DNA alterations based on the interaction with UvrA/UvrB

An enzyme-linked microplate immunoassay for the analysis of primary DNA les ions is described. The assay principle is based on the interaction of the b acterial DNA repair proteins UvrA and UvrB with DNA and on the immunodetect ion of UvrB forming a stable complex with covalently modified nucleotides. Using this technique we were able to detect damages in genomic DNA induced by uv light and by several different genotoxic agents. The detection sensit ivity of the method reaches down to the nanomolar range of the mutagenic co mpound depending on the type of the DNA alteration. The method might be use d in automated high-throughput studies. (C) 2001 Academic Press.