Microorganism Detection via ATP Level Changes
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current ATP bioluminescence methods struggle to detect microorganisms in biological samples containing cells due to background noise from cellular ATP, with existing techniques risking microorganism destruction and incomplete separation of cells and bacteria, and lacking reliable negative determination methods.
Innovation Solution
Culturing the sample in a microorganism culture medium to measure ATP level changes over time, allowing for the detection of microorganisms even in the presence of cells, and enabling negative determination by assessing ATP level decreases or increases, respectively.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If ATP bioluminescence method is used to detect microorganisms, then detection sensitivity and rapidity are improved, but cellular ATP creates background noise making microorganism detection difficult
Solution Approach 1:
The patent segments the detection process into two distinct phases: first measuring total ATP in the sample, then after selective cell lysis, measuring remaining ATP from microorganisms only. This temporal and procedural segmentation allows differentiation between cellular and microbial ATP signals, resolving the background noise problem while maintaining rapid detection capability.
Solution Approach 2:
The patent applies preliminary action by performing selective cell lysis treatment before the final microbial ATP measurement. This preliminary step removes the dominant cellular ATP background, enabling subsequent sensitive detection of microbial ATP without interference, thus improving measurement precision while preserving the rapidity advantage of ATP bioluminescence method.
2Measurement precision
If surfactant is used to selectively destroy cells, then cellular ATP background is removed, but microorganisms may be destroyed simultaneously
Solution Approach 1:
The patent applies local quality by using cell-specific lysis conditions that exploit the structural differences between eukaryotic cells and prokaryotic microorganisms. The lysis treatment is optimized to target eukaryotic cell membranes specifically while preserving prokaryotic cell integrity, achieving selective background removal without compromising microbial detection targets.
Solution Approach 2:
The patent utilizes parameter changes by carefully controlling lysis treatment parameters (such as enzyme concentration, incubation time, temperature) to create a window where eukaryotic cells are lysed but prokaryotic cells remain intact. This parameter optimization ensures selective cell destruction while maintaining microorganism viability for subsequent ATP measurement.
3Measurement precision
If filter method is used to separate cells and bacteria, then cellular ATP is removed, but cells smaller than filter pore size are not completely separated
Solution Approach 1:
The patent uses selective lysis treatment as an intermediary mechanism to achieve separation based on biological properties rather than physical size. This intermediary approach targets the biochemical differences between cell types, providing more reliable separation than physical filtration alone, especially for small cells that would pass through standard filters.
4Measurement precision
If microorganism is present in extremely small amount, then detection sensitivity is required, but ATP level from single microorganism is very low at approximately 1 amol
Solution Approach 1:
The patent extracts the microbial ATP signal from the overwhelming cellular background by removing cellular ATP through selective lysis. This extraction of the target signal from interfering background enables detection of extremely low concentrations (amol level) by eliminating the dominant background signal that would otherwise mask the微弱 microbial ATP.
Solution Approach 2:
The patent employs feedback by comparing ATP levels before and after selective lysis treatment. The difference in ATP signals provides feedback information about microbial presence, enabling detection of trace amounts through the change rather than relying on absolute low-level measurements, thus enhancing detection sensitivity.
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
This method effectively detects microorganisms and confirms sterility in samples like regenerative medical products and blood preparations, providing rapid and reliable results by distinguishing ATP changes associated with microorganism presence or absence.
Implementation Method 1
luciferase or luciferin which are chemiluminescent and produce light, in order to measure the luminescence produced by the reaction of the ATP in cells with luciferase and luciferin
Implementation Method 2
The ATP bioluminescence method is an enzymatic reaction, and thus, is excellent in sensitivity and rapidity
Data Source
AI summary
Provided is a method for detecting a microorganism in a test sample containing cells, including a step of adding the sample to a microorganism culture medium, a step of culturing the microorganism culture medium containing the sample, a step of sampling a part of the culture medium at a predetermined time, a step of acquiring the ATP level of the sampled culture medium, and a step of detecting the microorganism in the sample based on the change of the ATP level over time.