Lateral Flow Assay for Rapid Vibrio Detection
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Solution Overview
Problem
Current methods for detecting Vibrio bacteria in aquaculture products are time-consuming and require laboratory-based culture and PCR techniques, which are not suitable for rapid, on-site detection, posing challenges for food safety and aquaculture management.
Innovation Solution
Development of rapid paper- or substrate-based immunoassays that include test strips with detection zones for Vibrio species, activity, and virulence, utilizing antibodies and nanoparticles to provide quick and readable results, potentially using a cell phone for analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If culture method and MPN PCR method are used to detect Vibrio, then detection accuracy is improved, but detection time is significantly increased
Solution Approach 1:
The detection system is segmented into multiple functional zones on a single test strip: sample application zone, conjugate zone with antibody-coated particles, test zone with immobilized antibodies for Vibrio detection, and control zone. This segmentation allows simultaneous execution of multiple detection functions, reducing overall detection time while maintaining accuracy.
Solution Approach 2:
The patent combines multiple detection capabilities (Vibrio presence detection, activity detection via quorum sensing molecules, and virulence detection via TDH/TRH toxins) into a single integrated lateral flow assay platform. This merging eliminates the need for separate laboratory procedures, achieving rapid multi-parameter detection in one test.
2Reliability
If lab-based culture and PCR methods are used, then detection reliability is improved, but operational complexity and infrastructure requirements increase
Solution Approach 1:
The test strip performs self-diagnosis through built-in control zones that automatically verify proper fluid flow and reagent functionality. The assay includes self-contained conjugate reservoirs and pre-coated antibodies, eliminating the need for external laboratory equipment or complex operational procedures. Users simply apply sample and read results.
Solution Approach 2:
The patent transforms the detection parameters from requiring complex laboratory instrumentation to using simple visual or smartphone-readable color changes. The lateral flow assay converts molecular binding events into visible signal changes at test zones, making reliable detection accessible without specialized laboratory infrastructure.
3Measurement precision
If centralized lab processing is used, then detection precision is maintained, but sample processing speed and throughput are reduced
Solution Approach 1:
The patent transitions from two-dimensional laboratory workflow (sample preparation → incubation → analysis) to a linear one-dimensional lateral flow path where all reactions occur simultaneously as fluid moves through predefined zones. This dimensional simplification enables rapid parallel processing of multiple detection functions without sequential steps.
Solution Approach 2:
The test strip is pre-prepared with immobilized antibodies, conjugate reservoirs, and control mechanisms before use. This preliminary preparation eliminates time-consuming laboratory setup steps during actual detection, allowing immediate processing of samples at point-of-care locations with high throughput capability.
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, on-site detection of Vibrio bacteria, reducing turnaround time and improving food safety and aquaculture management by providing immediate results for the presence, activity, and virulence of pathogens.
Implementation Method 1
a porous carrier, e.g., nitrocellulose or filter-type paper... allows sample to flow from the sample pad through the first end of the test strip
Implementation Method 2
a wick, such as a material of sufficient absorbency to draw fluid through the test strip from the first end to the second end
Implementation Method 3
The first detection zone is configured to detect the presence of bacteria of the species Vibrio, and can include at least one antibody, e.g., a plurality of antibodies, that bind specifically to a specific kind of bacteria, e.g., of the Vibrio species
Implementation Method 4
The second detection zone is configured to determine whether any bacteria present in the sample is active, and comprises immobilized E. coli transformed with a quorum sensing plasmid specific to bacteria of the Vibrio species
Implementation Method 5
a chromogenic or luminescent substrate that is sensitive to the presence of AHL
Implementation Method 6
a chromogenic or luminescent substrate that is sensitive to the presence of AHL
Implementation Method 7
The third detection zone is configured to determine whether any bacteria present in the sample is virulent, and can include at least one antibody, e.g., a plurality of antibodies, that bind specifically to at least one of Thermostable Direct Hemolysin (TDH) and TDH-Related Hemolysin (TRH)
Data Source
AI summary
The present disclosure relates to methods, devices, assays and systems for rapid detection of food-borne pathogens, including Vibrios.