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Single bacterial cell detection using a mutant luciferase
Authors:Kenichi Noda  Tadahiro Matsuno  Hiroya Fujii  Tomonari Kogure  Masaaki Urata  Yasuo Asami  Akio Kuroda
Affiliation:(1) Interdisciplinary Research on Integration of Semiconductor and Biotechnology, Hiroshima University, 1-3-1 Kagamiyama, Higashihiroshima Hiroshima, 739-8530, Japan;(2) Bussan Nanotech Research Institute, 2-1 Kouyadai, Tsukuba Ibaraki, 305-0074, Japan
Abstract:
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.
Keywords:ATP assay  Bacteria detection  Mutant firefly luciferase
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