N-Assay ELISA for Rapid Pathogen Detection and Antimicrobial Susceptibility
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Solution Overview
Problem
Current methods for identifying clinically relevant pathogens and determining their antimicrobial susceptibility are time-consuming, resource-intensive, and often require specialized equipment, failing to provide rapid and accurate results, especially in cases of emerging antibiotic resistance.
Innovation Solution
A rapid enzyme-linked immunosorbent assay (ELISA) called N-Assay, which uses serial dilutions of microorganisms, specific antibodies, and enzyme-conjugated antibodies to detect and identify pathogens in as little as 30 minutes, with the ability to determine antimicrobial susceptibility in the same assay, reducing the overall time to 30 minutes or less compared to traditional methods.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If traditional culture methods are used for pathogen identification and antimicrobial susceptibility testing, then accuracy and reliability are maintained, but the time required increases to 3-5 days
Solution Approach 1:
The patent replaces traditional mechanical culture methods with an enzyme-linked immunosorbent assay (ELISA) system that uses antibody-antigen recognition and enzymatic reactions to detect pathogens and determine antimicrobial susceptibility. This substitution of biological recognition mechanisms with immunological and enzymatic processes enables rapid detection within hours rather than days.
Solution Approach 2:
The patent changes the detection parameters from traditional culture-based methods to immunological and enzymatic parameters. By measuring antibody binding and enzyme activity rather than bacterial growth, the system achieves rapid results while maintaining accuracy. The ELISA methodology transforms the detection parameter from temporal (growth time) to biochemical (antibody-antigen interaction).
2Loss of time
If nucleic acid amplification technology is used to identify pathogens, then detection speed increases, but the ability to determine antimicrobial susceptibility is lost
Solution Approach 1:
The patent merges pathogen identification and antimicrobial susceptibility testing into a single ELISA assay system. By combining both detection functions in one integrated platform, the system simultaneously provides rapid pathogen identification and susceptibility information without requiring separate tests, thus resolving the trade-off between speed and comprehensive reliability.
Solution Approach 2:
The ELISA system is designed with multi-functionality to perform both pathogen detection and antimicrobial susceptibility testing. The assay uses the same fundamental principle of antibody-antigen interaction to achieve multiple objectives, making the system universally applicable for both identification and susceptibility determination in a single test.
3Loss of time
If chromogenic broth/agar is used to reduce culture time, then initial detection speed improves, but specialized resources and training requirements increase
Solution Approach 1:
The patent employs disposable microplate wells and pre-coated antibodies that can be easily discarded after use. This approach eliminates the need for expensive, complex chromogenic broth/agar systems and specialized equipment. The ELISA microplate format uses simple, single-use components that reduce both initial investment and ongoing operational complexity.
Solution Approach 2:
The patent introduces enzyme-conjugated antibodies as intermediaries to detect pathogen-antibody complexes. This intermediary mechanism simplifies the detection system compared to chromogenic substrates, requiring only standard laboratory equipment rather than specialized chromogenic imaging systems, thus reducing resource and training requirements.
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 N-Assay provides a highly sensitive and specific method for identifying microorganisms and determining their susceptibility to antibiotics in a fraction of the time required by traditional methods, enabling more targeted treatment approaches and reducing the risk of antibiotic resistance.
Implementation Method 1
adding an enzyme-conjugated antibody specific for the microorganism to the plurality of wells and incubating the plurality of wells for about 7 minutes
Implementation Method 2
adding a substrate specific to the enzyme conjugated to the antibody to the plurality of wells and incubating the plurality of wells for about 3 minutes; detecting and identifying the microorganism in wells showing a yellow color
Data Source
AI summary
The present invention provides a rapid, highly sensitive and specific enzyme-linked immunosorbent assay (ELISA), referred to as N-Assay, a device, and a kit, for detection and identification of microorganisms in a sample, in thirty minutes or less, with little or no interference from non-target microorganisms. The present invention also provides for simultaneous determination of antimicrobial susceptibility of microorganism in the N-Assay.