Microbial Drug Resistance Evaluation via Enzyme Extraction and MS
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Solution Overview
Problem
Existing methods for evaluating antimicrobial resistance in bacteria require culturing the bacteria in the presence of antimicrobial agents, which is time-consuming.
Innovation Solution
A method involving an extraction step to obtain a drug-digested enzyme from a microorganism, a mixing step with a drug, followed by mass spectrometry to detect degradation products, and an evaluation step to determine resistance based on the presence or absence of peaks in the mass spectrum.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If bacteria are cultured in the presence of antimicrobial agents to evaluate resistance, then the evaluation can detect metabolic changes and degradation products, but the process takes several hours due to the culturing requirement
Solution Approach 1:
The patent applies preliminary action by pre-extracting enzymes from bacteria and storing them for later use. Instead of culturing bacteria at the time of evaluation, the extraction step is performed in advance to obtain enzyme-containing extracts, which can then be directly mixed with antimicrobial agents and analyzed by mass spectrometry without requiring time-consuming culturing procedures.
Solution Approach 2:
The patent applies extraction by separating the enzyme component from the whole bacterium. The extraction step isolates drug-digested enzymes from bacterial cells, creating a concentrated enzyme extract that can be used directly in resistance evaluation. This extraction eliminates the need to maintain live bacteria during the evaluation process, thereby reducing evaluation time while preserving detection capability.
2Adaptability or versatility
If mass spectrometry is performed on metabolites using LC-MS, then metabolite detection is achieved, but using the same MALDI-MS for both causative agent identification and resistance evaluation requires additional method development
Solution Approach 1:
The patent applies universality by adapting the MALDI-MS apparatus to perform dual functions: identifying causative agents and evaluating antimicrobial resistance. The same mass spectrometer used for bacterial identification is configured to detect degradation products of antimicrobial agents, eliminating the need for separate LC-MS equipment and enabling one instrument to serve multiple diagnostic purposes.
Solution Approach 2:
The patent applies parameter changes by modifying the mass spectrometry parameters and sample preparation methods to suit resistance evaluation. The MALDI-MS parameters are optimized to detect specific degradation products of antimicrobial agents, and the extraction method is adjusted to produce enzyme extracts suitable for MALDI analysis, thereby adapting the apparatus for its second function.
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
Enables rapid evaluation of antimicrobial resistance without the need for culturing, allowing for quick identification of drug-resistant bacteria.
Implementation Method 1
β-lactamase hydrolyzes the β-lactam ring of β-lactam antimicrobial agents to inactivate them
Implementation Method 2
the pattern of the obtained mass spectrum is collated with the mass spectrum patterns of a large number of known bacteria previously registered in a database
Data Source
AI summary
[Problem] To provide a method for rapidly evaluating the resistance of microorganisms such as bacteria to a drug.[Solution] The method comprises an extraction step of performing an operation to obtain an extract containing a drug-degrading enzyme that can be produced by the microorganism from a sample containing the microorganism; a mixing step of mixing the extract and a drug to obtain a mixture; an analysis step of subjecting the mixture to mass spectrometry; and an evaluation step of determining whether the microorganism is resistant to the drug by detecting the presence or absence of a peak derived from a degradation product of the drug from the mass spectrum obtained in the analysis step.