Development of a new bioluminescent mutagenicity assay based on the Ames test.
A newly developed rapid mutagenicity assay based on the adenosine triphosphate (ATP)-bioluminescence technique and the Ames test is described. Salmonella typhimurium strains TA98 and TA100 were exposed in an appropriate liquid medium to the direct mutagens 4-nitroquinoline-N-oxide and methyl methanesulphonate, respectively, and to the indirect mutagen 2-aminoanthracene. Both auxotrophic and prototrophic growth were monitored throughout the incubation period as variations in the intracellular ATP levels by means of the luciferin-luciferase assay. After 9-12 h of incubation a dose-response increase in the levels of ATP was readily detected. In order to demonstrate that this increase was due to the growth of revertant bacteria, aliquots from each culture were plated on minimal agar plates. A very good correlation between the changes in ATP levels and the appearance of revertant colonies on the plates was found. Given the rapidity of this method as compared with conventional mutagenicity assays, it has potential for industrial and environmental applications. Other potential applications are also discussed.
Guadaño
A
A
Centro de Ciencias Medioambientales, CSIC, c/Serrano 115 dpdo, 28006 Madrid and R&D Laboratory, Gomensoro/Biotech, Madrid, Spain. aguadano@ccma.csic.es
de la Peña
E
E
González-Coloma
A
A
Alvarez
J F
JF
D015112:4-Nitroquinoline-1-oxide / Q000633:toxicity; D000255:Adenosine Triphosphate / Q000378:metabolism; D000873:Anthracenes / Q000633:toxicity; D008741:Methyl Methanesulfonate / Q000633:toxicity; D009152:Mutagenicity Tests / Q000379:methods*; D009153:Mutagens / Q000633:toxicity; D012486:Salmonella typhimurium / Q000235:genetics / Q000254:growth & development
D000873:Anthracenes; D009153:Mutagens; D015112:4-Nitroquinoline-1-oxide; C006593:2-anthramine; D000255:Adenosine Triphosphate; D008741:Methyl Methanesulfonate