Multiplexed Droplet Particle Detection via Degradable Carrier Release
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Solution Overview
Problem
Current methods for multiplexed analysis of multiple targets in a test sample are limited in their ability to efficiently detect and differentiate between various nucleic acid sequences, bacterial types, antibiotic resistance, and antigens within a single assay, often requiring multiple tests and lacking sensitivity and specificity.
Innovation Solution
A system and method utilizing sets of particles that are degradable in response to specific environmental conditions, each set containing target-specific reagents and optically detectable identifiers, combined with a reporter probe that emits distinct wavelengths upon reaction, allowing for simultaneous detection and differentiation of multiple targets in aqueous-in-oil reaction droplets.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If multiple separate tests are used to detect different nucleic acid sequences, then detection coverage is improved, but assay complexity increases
Solution Approach 1:
The patent combines multiple detection assays into a single multiplexed reaction by co-incubating different primer sets and reporter probes targeting multiple nucleic acid sequences simultaneously. This merging approach allows detection of multiple targets in one assay, improving detection coverage while reducing overall assay complexity compared to running separate tests.
Solution Approach 2:
The patent creates a universal detection platform that can simultaneously detect multiple different nucleic acid sequences using a common reaction system. The system employs universal reaction conditions and detection mechanisms that work across multiple targets, allowing one assay to perform the function of multiple separate assays.
2Productivity
If multiple targets are detected simultaneously, then productivity is improved, but measurement precision deteriorates
Solution Approach 1:
The patent assigns distinct local qualities to different detection channels by using reporter probes that emit at different wavelengths or have different detection characteristics. Each target is detected with specific reagents and detection parameters optimized for that target, allowing simultaneous detection while maintaining individual measurement precision through channel-specific optimization.
Solution Approach 2:
The patent resolves measurement precision issues by adding spectral dimensionality to the detection system. Different targets are distinguished not just by presence/absence but by their unique spectral signatures (wavelengths), allowing simultaneous detection of multiple targets with high specificity through multi-dimensional signal resolution.
3Stability of the object's composition
If degradable particles are used to release reagents, then reagent stability is improved, but device complexity increases
Solution Approach 1:
The patent applies preliminary action by pre-encapsulating reagents within degradable particles before the assay begins. The particles are prepared in advance with reagents in a stable, protected state, and only release their contents when exposed to specific degradation triggers during the assay, thereby maintaining reagent stability until needed while enabling controlled release.
Solution Approach 2:
The degradable particles serve as intermediaries that mediate between reagent storage and reagent release. These particles protect reagents during storage and transport, then facilitate controlled release under specific assay conditions, acting as a bridge between the need for reagent stability and the need for reagent availability.
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 efficient and sensitive multiplexed analysis of multiple targets by stochastic distribution and reaction within droplets, providing clear optical signals for each target, thereby enhancing detection capabilities and reducing assay complexity.
Implementation Method 1
each particle of the first set of particles degradable in response to a first environmental condition
Implementation Method 2
first optically detectable identifier capable of emitting a first wavelength
Implementation Method 3
second optically detectable identifier capable of emitting a second wavelength
Implementation Method 4
at least one optically detectable reporter probe capable of constitutively emitting a third wavelength in response to amplification
Implementation Method 5
aqueous-in-oil reaction droplets
Data Source
AI summary
Described herein are systems and methods for multiplexed analysis of two or more targets in a test sample including a first set of particles including a first set of target-specific reagents and a first optically detectable identifier capable of emitting a first wavelength indicative of a first target, and at least one second set of particles including a second set of target-specific reagents and a second optically detectable identifier capable of emitting a second wavelength indicative of a second target; and at least one optically detectable reporter probe capable of constitutively emitting a third wavelength in response to reaction of the first set of target-specific reagents with the first target in the test sample and/or reaction of the second set of target-specific reagents with the second target in the test sample, wherein the first wavelength, the second wavelength, and the third wavelength are optically discernable from one another.