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Related Experiment Video

Updated: Mar 10, 2026

'Bioluminescent' Reporter Phage for the Detection of Category A Bacterial Pathogens
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Attomol-level ATP bioluminometer for detecting single bacterium.

Masahiro Okanojo1, Noe Miyashita2, Aya Tazaki2

  • 1Central Research Laboratory, Hitachi Ltd, Tokyo, Japan.

Luminescence : the Journal of Biological and Chemical Luminescence
|December 14, 2016
PubMed
Summary
This summary is machine-generated.

We created a highly sensitive automated ATP bioluminometer capable of detecting single bacteria. This advanced instrument offers ultra-high sensitivity for measuring adenosine triphosphate (ATP) luminescence from minimal bacterial samples.

Keywords:
attomol-level ATPautomated luminometerautomated washingrapid detectionreaction tubes

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Microbiology

Background:

  • Accurate quantification of microbial populations is crucial in various scientific fields.
  • Traditional methods for bacterial detection can be time-consuming and lack sensitivity.
  • Adenosine triphosphate (ATP) bioluminescence offers a rapid and sensitive method for assessing viable biomass.

Purpose of the Study:

  • To develop and validate an automated, high-sensitivity ATP bioluminometer for single-bacterium detection.
  • To assess the performance of the instrument in terms of background noise, linearity, and detection limits.
  • To establish the relationship between intracellular ATP levels and Colony Forming Units (CFU) for key bacterial species.

Main Methods:

  • Design and construction of an automated system incorporating a tube rack, washing baths, XYZ actuators, and a high-sensitivity optical system.
  • Optimization of reaction tubes to minimize background signal intensities.
  • Calibration of the ATP bioluminometer using known ATP concentrations and validation with Escherichia coli and Staphylococcus aureus cultures.

Main Results:

  • The developed ATP bioluminometer achieved a low background signal intensity of 18 Relative Light Units (RLU).
  • A linear ATP calibration curve was obtained from 0 to 5 amol, with a detection limit of 0.8 amol.
  • Linear relationships were established between intracellular ATP and CFU for E. coli (3.3 amol/CFU) and S. aureus (1.6 amol/CFU).

Conclusions:

  • The automated ATP bioluminometer demonstrates ultra-high sensitivity for detecting and quantifying bacterial ATP.
  • The instrument is a powerful tool for microbial analysis, enabling the measurement of ATP luminescence from very small bacterial numbers.
  • This technology has significant potential for applications in diagnostics, environmental monitoring, and food safety.