Microplate Enzyme-Linked Immunosorbent Assay for the Detection of Primary DNA Alterations Based on the Interaction with UvrA/UvrB

✍ Scribed by Guido Böse; Jürgen Kuhlmann; Martina Plass; Oliver Müller

• Publisher: Elsevier Science
• Year: 2001
• Tongue: English
• Weight: 141 KB
• Volume: 292
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.2001.5054

✦ Synopsis

An enzyme-linked microplate immunoassay for the analysis of primary DNA lesions is described. The assay principle is based on the interaction of the bacterial DNA repair proteins UvrA and UvrB with DNA and on the immunodetection 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 sensitivity of the method reaches down to the nanomolar range of the mutagenic compound depending on the type of the DNA alteration. The method might be used in automated high-throughput studies.