Antimicrobial Susceptibility Assay via Live Cell Reporter
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Solution Overview
Problem
Current methods for determining the mechanisms of antimicrobial susceptibility in organisms are complex, costly, and require multiple assays, as they cannot simultaneously identify organisms, determine phenotypic susceptibility, and identify mechanisms involved in susceptibility phenotypes, necessitating the use of multiple systems and techniques.
Innovation Solution
A method involving a live cell reporter system combined with inhibitors and antimicrobial agents to detect the presence or absence of mechanisms of antimicrobial resistance by monitoring cell viability and proliferation, using non-replicative transduction particles and caged substrates to determine if resistance mechanisms are expressed in microorganisms.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of information
If multiple separate assays and systems are used to identify organisms, determine phenotypic susceptibility, and identify resistance mechanisms, then comprehensive information can be obtained, but the complexity, cost, and time required increase significantly
Solution Approach 1:
The patent combines organism identification, phenotypic susceptibility determination, and resistance mechanism identification into a single integrated assay system. This merging of multiple functions into one system eliminates the need for separate assays, reducing complexity while maintaining comprehensive information acquisition.
Solution Approach 2:
The assay system is designed to perform multiple functions simultaneously: identifying the organism, determining its susceptibility phenotype, and detecting the specific resistance mechanism. This multi-functional approach allows a single system to replace multiple specialized systems.
2Measurement precision
If traditional culture techniques and multiple assays are used to determine resistance mechanisms, then accurate mechanism identification can be achieved, but the time and resource requirements increase
Solution Approach 1:
The system performs preliminary actions by simultaneously conducting organism identification and susceptibility testing in the same assay. The live cell reporter system allows real-time monitoring of susceptibility phenotype while the inhibitor-based approach concurrently identifies resistance mechanisms, eliminating sequential testing steps.
3Loss of information
If NAA techniques are used to detect resistance genes, then gene presence can be identified, but the actual expression and susceptibility cannot be determined
Solution Approach 1:
The patent uses an intermediary approach by introducing inhibitors that specifically target resistance mechanisms. The inhibitors act as mediators between the resistance mechanism and the susceptibility phenotype, allowing the system to detect not only the presence of resistance genes but also their actual expression and functional impact on susceptibility.
4Measurement precision
If colorimetric assays are used to detect enzyme expression, then enzymatic mechanisms can be identified, but organism identification and non-enzymatic mechanisms cannot be determined
Solution Approach 1:
The assay system is designed with universal applicability to detect multiple types of resistance mechanisms beyond just enzymatic ones. By combining live cell reporter-based susceptibility phenotyping with inhibitor-based mechanism identification, the system can detect enzymatic, non-enzymatic, and combination mechanisms across different organism types.
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 approach allows for a single assay to identify organisms, determine susceptibility, and identify resistance mechanisms, reducing the need for multiple systems and techniques, thereby simplifying and cost-effectively determining antimicrobial susceptibility.
Implementation Method 1
a live cell reporter capable of generating a detectable signal to monitor cell viability and proliferation
Data Source
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AI summary
Disclosed herein are methods and compositions for determining the presence or absence of a mechanism of antimicrobial resistance in a sample.