ATP-Diphosphohydrolase Kit for Bacterial ATP Detection
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Solution Overview
Problem
Traditional microbiological tests for detecting microbial contamination in industrial samples are slow due to the need for incubation periods of 24-48 hours or more, especially for stressed or slow-growing microorganisms, and there is a need for a method to distinguish microbial ATP from non-microbial ATP in samples like milk.
Innovation Solution
Aqueous compositions comprising ATP-diphosphohydrolase, polyols, buffer reagents, and proteins that are stable at ambient temperatures, allowing for the retention of greater than 90% of initial enzyme activity for extended periods, enabling rapid detection of microbial ATP through bioluminescence.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional culture techniques are used to detect microbial contamination, then the detection is accurate and reliable, but the testing time is extended to 24-48 hours or more
Solution Approach 1:
The detection process is segmented into two distinct phases: (1) rapid ATP bioluminescence screening that provides immediate results for decision-making, and (2) traditional culture confirmation that ensures accuracy. This segmentation allows the system to deliver fast initial results while maintaining reliable verification through conventional methods.
Solution Approach 2:
ATP bioluminescence serves as an intermediary screening method between sample collection and final culture confirmation. It provides a rapid intermediate assessment that can guide whether further culture testing is necessary, reducing the overall time loss while preserving detection reliability through the intermediary step.
2Productivity
If ATP bioluminescence reaction is used to detect microbial ATP, then the testing speed is improved, but the ability to distinguish microbial ATP from non-microbial ATP is compromised
Solution Approach 1:
The invention extracts and removes non-microbial ATP from the sample matrix before performing the ATP bioluminescence reaction. By selectively eliminating ATP from non-microbial sources (such as plant tissue, food matrices, or environmental samples), the assay achieves both rapid testing speed and high specificity for microbial detection.
Solution Approach 2:
An intermediary treatment step is introduced between sample collection and ATP measurement that selectively removes or neutralizes non-microbial ATP. This intermediary process preserves the rapid speed of ATP bioluminescence while enhancing measurement precision by eliminating false positive signals from non-microbial sources.
3Ease of operation
If ATP-diphosphohydrolase is stored in aqueous solution at room temperature, then the ease of operation is improved, but the enzyme activity is lost over time
Solution Approach 1:
The invention employs a disposable lyophilized enzyme formulation that is stable at room temperature but intended for single-use or limited-use applications. The enzyme is provided in a stable, dried state that requires no refrigeration, and is activated by adding the sample or buffer immediately before use, ensuring both ease of operation and reliability of enzyme activity.
Solution Approach 2:
The physical state of the enzyme is changed from aqueous solution to lyophilized (dried) form, which fundamentally alters its stability parameters. The lyophilized enzyme can be stored at room temperature indefinitely without loss of activity, and the enzyme is reconstituted in buffer or sample immediately before use, combining storage convenience with activity reliability.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The stabilized ATP-diphosphohydrolase compositions maintain enzyme activity for at least 28 days at room temperature, facilitating the detection of microbial ATP in samples by effectively hydrolyzing non-microbial ATP and allowing for quick measurement of microbial presence, thus speeding up the testing process.
Implementation Method 1
ATP-diphosphohydrolase (apyrase) that catalyzes the hydrolysis of ATP to AMP and inorganic phosphate
Implementation Method 2
When adenosine-5'-triphosphate (ATP) is added to this reagent, luciferase catalyzes the emission of light
Data Source
AI summary
A kit for detecting bacterial ATP in a sample is provided. The kit comprises an aqueous composition having a pH of about 6.0 to 7.2. The aqueous composition comprises effective amounts of a polyol, a buffer reagent, a protein, and ATP-diphosphohydrolase. A method of using the kit to detect bacterial ATP is also provided.