Sealed ATP Cell Detection with Intermittent Reagent Contact
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Solution Overview
Problem
Existing methods for detecting microbes in specimens, such as fluorimetry and ATP methods, suffer from low sensitivity and potential contamination, especially when dealing with low initial microbial counts or slow proliferating microbes, leading to prolonged detection times and false positives.
Innovation Solution
A cell detector and method that uses a sealing container with separate culture and luminescent reagent compartments, controlled contact through a mechanism, and a photodetector to measure luminescence changes over time, enhancing sensitivity and reducing detection time.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If the culture medium and luminescent reagent are continuously mixed, then the luminescence detection can be performed continuously, but the ATP is always consumed in the luminous reaction causing decreased sensitivity and detection precision
Solution Approach 1:
The patent implements periodic action by intermittently mixing the culture medium and luminescent reagent instead of continuous mixing. The mixing is performed at specific time intervals (e.g., every hour), allowing ATP to accumulate during non-mixing periods and be measured during mixing periods. This periodic approach prevents complete ATP consumption while maintaining continuous detection capability over extended periods.
Solution Approach 2:
The patent applies preliminary action by pre-mixing the luminescent reagent with the culture medium before the actual detection process. This preliminary mixing allows the system to be prepared in advance, and the actual ATP measurement occurs only when needed, preventing unnecessary ATP consumption while maintaining readiness for continuous detection.
2Productivity
If the culture medium is fractionated and mixed with luminescent reagent every hour, then the microbial proliferation can be monitored, but the operation increases contamination risk leading to false positive results
Solution Approach 1:
The patent merges the culture medium and luminescent reagent into a single sealed container system. The luminescent reagent is pre-added to the culture medium in the same container, eliminating the need for separate fractionation and mixing operations. This integration maintains the ability to monitor microbial proliferation by measuring luminescence, while simultaneously preventing contamination that would occur during repeated opening and mixing operations.
Solution Approach 2:
The system performs self-service by automatically measuring luminescence within the sealed container without requiring external intervention for fractionation or mixing. The luminescent reagent remains in the sealed container with the culture medium, and measurements are taken directly in this sealed environment, eliminating the need for operator intervention that could introduce contamination.
3Ease of operation
If fluorimetry is used to detect microbial proliferation, then the detection method is simple, but the sensitivity is low requiring 10^7 CFU/mL or more to detect microbes
Solution Approach 1:
The patent changes the detection parameter from fluorimetry to luminescence measurement. By using a luminescent reagent that produces light through chemical reaction with ATP, the system achieves much higher sensitivity (detecting 100 CFU or more) while maintaining operational simplicity. The luminescence intensity directly correlates with ATP concentration, providing both high sensitivity and ease of measurement.
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 method achieves enhanced detection sensitivity for microbes, reducing detection time and minimizing contamination risks, allowing for rapid and accurate microbial proliferation detection.
Implementation Method 1
a luminescent reagent portion containing a luminescent reagent that emits light upon contact with the culture medium
Data Source
AI summary
A cell detector and detecting method enhance detection sensitivity for microbes according to the ATP method and reduce the detection period. The cell detector includes a sealing container having a culture portion for containing a culture medium containing the specimen, a sealable specimen introducing portion, and a luminescent reagent portion that emits light upon contact with the culture medium, a contact mechanism for controlling the contact between the culture medium and the luminescent reagent portion, a photodetector for detecting a luminescence resulting from the contact between the culture medium and the luminescent reagent portion, and a calculation unit for calculating a luminescent quantity from detection signals of the photodetector to determine cell proliferation on the basis of change in the luminescent quantity over time. The culture portion and the luminescent reagent portion are separately positioned, and the contact mechanism intermittently brings the culture medium and the luminescent reagent into contact.