Single bacterial cell detection using a mutant luciferase
Previously, we constructed a genetically modified luciferase of Photinus pyralis that generates more than 10-fold higher luminescence intensity than the wild-type enzyme. In this study, we demonstrate that this modified luciferase enables us to detect ATP at 10−18 mol, which is almost equal to the quantity contained in a single bacterial cell. Consequently,we have been able to detect bacterial contamination in samples as low as one colony-forming unit (c.f.u.) for both Escherichia coli and Bacillus subtilis.
KeywordsATP assay Bacteria detection Mutant firefly luciferase
- Trudil D, Loomis L, Pabon R, Hasan JAK, Trudil CL (2000) Rapid ATP method for the screening and identification of bacteria in food and water samples. Biocatal 41:27–29Google Scholar