Microbial ATP extraction and detection system
AssigneePromega Corporation
InventorsFrank Fan, Braeden Butler, Keith V. Wood
Life of this patent
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Abstract
The present invention is directed to compositions and methods for single-step extraction and detection of ATP levels from microbial cells. The disclosed compositions are formulated to efficiently elicit bioluminescent detection of ATP among a broad variety of different microorganisms using a common single-step reagent composition. Additional luminescence-based methods are provided for identifying other useful extracting agents or for screening compounds for their pharmaceutical or biological effects on microbial cells.
Claims26
A composition for extracting ATP from a gram negative microorganism comprising:
- (a) a reaction buffer;
- (b) one or more ATP extracting agents;
- (c) a divalent cation at a first concentration;
- and
- (d) a divalent cation chelator at a second concentration;
- wherein the first concentration is at least 10 mM and the second concentration is at least about equal to or greater than the first concentration.
An aqueous composition for detecting ATP in a sample suspected of containing a microorganism comprising:
- (a) a reaction buffer;
- (b) one or more ATP extracting agents;
- (c) a divalent cation at a first concentration
- (d) a divalent cation chelator at a second concentration;
- and
- (e) a luciferase enzyme
- wherein the second concentration is at least 10 mM and the difference between the first concentration and the second concentration is less than about 5 mM;
- and
- wherein the one or more ATP extracting agents comprises a compound or combination of compounds capable of disrupting the integrity of a bacterium or yeast to effect release of ATP therefrom.
An aqueous composition for detecting ATP in a sample suspected of containing a microorganism comprising:
- (a) a reaction buffer;
- (b) one or more ATP extracting agents;
- (c) a divalent cation at a first concentration;
- (d) a divalent cation chelator at a second concentration;
- wherein the second concentration is greater than the first concentration. wherein the first concentration is at least 10 mM and the difference between the first concentration and the second concentration is less than about 5 mM;
- and
- wherein the one or more ATP extracting agents comprises a compound or combination of compounds capable of disrupting the integrity of a bacterium or yeast to effect release of ATP therefrom.
Dependencies are inferred from the claim text.