What is ATP?

Atp is Adenosine Triphosphate molecule produced by all living cells, and is used to store or free energy for intra-cellular or extra-cellular reactions.

When cells are dying and their membranes are degrading, Atp is released from the cell

Intra-cellular Atp production if detected, should confirm that cells present in the sample were still alive.

Atp is produced by any cells as skin hair and other somatic cells, plant cell, bacteria, protozoa, pollens, micro-algae, free Atp molecules can also be released from dead cells. So detection of total Atp does not bring any information on the type of cell present.

How do we detect ATP?

For bioluminescence reaction cofactor Mg++  must not bind to other ions prior to bioluminescence reaction as it may happen with metal ions, fulvic and humic acids as other chelatants, salts. Others biocides may also destroy luciferase enzyme or change its 3 D protein conformation prior to analysis.

If these substances are not flushed prior to intracellular Atp extraction, then it is quenching bioluminescence signal, and may reduce or completely inhibit completely bioluminescence signal.

Limitations of classic ATPbioluminescence technologies:

False positives:

  • Bacterial ATP is not discriminated from total ATP
  • Non bacterial ATP exceeds largely bacterial ATP values:
    • ATp on only one skin cell, somatic cells like blood... > 1000 bacteria
  • triggers unnecessarily false positives (false alerts)
  • therefore not correlated to sensitive plate count mtehods

False negatives:

  • no flushing of inhibiting substances before detecting Atp

  • Bioluminescence reaction is reduced even destroyed by inhibiting substances present in samples

    • bioluminescen signal reduced > 90% (Velasquez et al, 1997)

  • false negatives: quenched reaction let user believe that sample is with low bacteria count

  • Inhibiting factor unpredictable from one environmental sample to the next one:

    • number and concentration of unidentified inhibiting substances varies from sample to sample

Lack of sensitivity of Atpbioluminescence due to small volume analyzed and 100 fold dilution factor by luer lock syringe Atp extraction method:

  • Low volume sampled by swabbing (0.1ml sampled is not representative of the bacterial population present in liquids)
  • Dilution factor of kits using syringe filters to concentrate total cells contents of samples use very agressive reagent to lyse the cells entrapped inside the filter and release its intracellular ATP, however the Atp extractant used if not diluted will destroy the substrate-enzyme and quench the bioluminescence signal.
  • only aliquot of total Atp is used inside a luminometer using a transfer cuvette, with a maximum volume of 100 microliters (as shown down below drawings)


For Atpbioluminescence detection tools to be correlated to sensitive plate count methods or rapid viable bacteria detection methods :

  1. there must be a preliminary flushing of inhibiting substances present in the samples
  2. a selective non-bacterial ATP extraction and flushing
  3. flushing free Atp present in the sample
  4. selective bacterial ATP and its detection by bioluminescence
  5. to be able to reactive bacterial spores prior to its detection

To reply to these needs a world patented solution exists :

PROFILE-1 is the only quantitative bacterial Atpbioluminescence diagnostic test kit:

  • correlated with sensitive plate count methods and published in world-reputed scientific journals
  • not influenced by quenching substances through a patented flushing system
  • discriminating bacterial Atp from non-microbial ATP
  • detecting also bacteria spores (with an inventive reactivation protocol).
  • not diluting its extracted Atp prior to bioluminescence reaction