Chemiluminometric biochemical induction assay (CBIA) for the detection of DNA-damaging agents

• Published in: Journal of microbiological methods 2005-05, Vol.61 (2), p.277-280
• Main Authors: Singh, Maya Prakash, Arias, Daniel A., Greenstein, Michael
• Format: Article
• Language: English
• Subjects: Anti-Bacterial Agents - pharmacology; Bacteriology; beta-Galactosidase - analysis; Biological and medical sciences; CBIA; DNA Damage; DNA, Bacterial - drug effects; DNA-damaging; Escherichia coli - drug effects; Escherichia coli - genetics; Escherichia coli - metabolism; Fundamental and applied biological sciences. Psychology; Luminescent Measurements - methods; Microbiology; Miscellaneous; Screen
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/j.mimet.2004.11.011

A microbroth chemiluminometric version of the biochemical induction assay (BIA) was developed using a chemiluminescent substrate widely used to detect β-galactosidase in high-throughput screening (HTS) laboratories. The assay was run in both 96-well and 384-well plate formats using the Zymark RapidPlate® liquid handling system to transfer samples and reagents. Chemiluminescence was read using the Victor-2 multilabel counter. The new microbroth chemiluminometric method, the CBIA, allowed rapid screening of samples, crude extracts, and pure compounds for their DNA-damaging effects in bacteria. In screening a small subset of our natural products library samples by the agar plate BIA and the CBIA, the latter yielded a higher hit rate, suggesting it is more sensitive than the agar plate assay. The CBIA was unaffected by the colored samples often encountered during screening of crude natural products extracts.