Development of a genotoxicity detection system using a biosensor.

Abstract

The umu-lux test is a genotoxicity test using the two genetically modified S. typhmurium TA1535 strains (TL210 and TL210ctl) transformed with the luxCDABE (luciferase gene and fatty acid reductase genes) of Vibrio fischeri as a reporter gene. The TL210 strain detects genotoxicants and the TL210ctl strain detects cytotoxicants. In order to develop a highly sensitive, simple and rapid genotoxicity detection system, we constructed a biosensor using these immobilized strains. The biosensor consists of two immobilized microbial membranes, a sample vessel and photodetectors, and the genotoxicity detection system consists of the biosensor, an isothermal box, a photodetector and an air pump. The total measurement time for genotoxicants using this detection system is about 4 h. When 2% (v/v) DMSO was used as a control, the TL210 strain was not emitting light while the TL210ctl strain was. When 0.3 mg/l 4NQO was used as a genotoxicant, TL210 strain and TL210ctl strain were both emitting light. When HgCl2 was used as a cytotoxicant, neither the TL210 strain nor the TL210ctl strain were emitting light. Therefore, the false negative prevention function of a biosensor using the TL210ctl strain has been checked. These results show that our proposed system can correctly detect genotoxicants.