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.
Noda
Kenichi
K
Interdisciplinary Research on Integration of Semiconductor and Biotechnology, Hiroshima University, 1-3-1 Kagamiyama, Higashihiroshima, Hiroshima, 739-8530, Japan. noda@hiroshima-u.ac.jp
Matsuno
Tadahiro
T
Fujii
Hiroya
H
Kogure
Tomonari
T
Urata
Masaaki
M
Asami
Yasuo
Y
Kuroda
Akio
A
D000251:Adenosine Triphosphatases / Q000032:analysis; D001412:Bacillus subtilis / Q000302:isolation & purification*; D001431:Bacteriological Techniques; D015169:Colony Count, Microbial; D004926:Escherichia coli / Q000302:isolation & purification*; D049409:Luciferases, Firefly / Q000032:analysis*; D008163:Luminescent Measurements; D016297:Mutagenesis, Site-Directed
D049409:Luciferases, Firefly; D000251:Adenosine Triphosphatases