Single bacterial cell detection using a mutant luciferase

Biotechnol Lett. 2008 Jun;30(6):1051-4. doi: 10.1007/s10529-008-9643-3. Epub 2008 Jan 26.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / analysis
  • Bacillus subtilis / isolation & purification*
  • Bacteriological Techniques
  • Colony Count, Microbial
  • Escherichia coli / isolation & purification*
  • Luciferases, Firefly / analysis*
  • Luminescent Measurements
  • Mutagenesis, Site-Directed

Substances

  • Luciferases, Firefly
  • Adenosine Triphosphatases