Multiplexed Digital Assay Combinatorial Signal Detection

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Solution Overview

Problem

Current digital assays face limitations in multiplexing capabilities due to the high cost and impracticality of increasing the number of distinguishable dyes and detection channels, which restricts the ability to detect multiple targets simultaneously, especially in applications with limited sample volumes.

Innovation Solution

A color-based approach for multiplexed digital amplification assays that uses combinatorial signaling, where multiple targets are amplified in the same partitions, and composite signals are generated across different wavelength regimes to detect the presence of multiple targets using fewer detection channels, allowing for the calculation of individual target concentrations.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If the number of distinguishable dyes and detection channels is increased to detect more targets, then the multiplexing capability is improved, but the cost and practicality deteriorate

Engineering Contradiction:
Improvemultiplexing capabilityVSAvoidnumber of detection channels
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent combines multiple detection channels by using spectral unmixing algorithms that analyze composite fluorescence signals containing contributions from multiple dyes. Instead of requiring separate detection channels for each dye, the system merges the detection into fewer channels and uses computational methods to resolve the individual dye signals based on their spectral characteristics.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The patent transitions from spatial separation of detection channels to spectral dimension analysis. By measuring fluorescence intensities across multiple wavelengths in fewer detection channels and applying spectral unmixing, the system extracts information about multiple dyes without requiring an equal number of physical detection channels.

Inventive Principle:
Principle #17Another dimension (Dimensionality change)

2Adaptability or versatility

If the number of distinguishable dyes is increased to detect more targets, then the multiplexing capability is improved, but the cost and practicality deteriorate

Engineering Contradiction:
Improvemultiplexing capabilityVSAvoidcost of dyes and instruments
Core Design Contradiction:
Adaptability or versatilityVSEase of manufacture

Solution Approach 1:

The patent combines multiple detection channels by using spectral unmixing algorithms that analyze composite fluorescence signals containing contributions from multiple dyes. Instead of requiring separate detection channels for each dye, the system merges the detection into fewer channels and uses computational methods to resolve the individual dye signals based on their spectral characteristics.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The patent uses reference spectral data from individual dyes to create a computational model that can deconvolve composite signals. By having spectral fingerprints of individual dyes stored as references, the system can mathematically separate the contributions of multiple dyes in a composite signal, effectively creating virtual detection channels through computational copying of spectral information.

Inventive Principle:
Principle #26Copying

3Measurement precision

If more detection channels are used to detect more colors, then the accuracy of target detection is improved, but the cost and complexity increase

Engineering Contradiction:
Improvetarget detection accuracyVSAvoidnumber of detection channels
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent transitions from spatial separation of detection channels to spectral dimension analysis. By measuring fluorescence intensities across multiple wavelengths in fewer detection channels and applying spectral unmixing, the system extracts information about multiple dyes without requiring an equal number of physical detection channels.

Inventive Principle:
Principle #17Another dimension (Dimensionality change)

Solution Approach 2:

The patent employs iterative spectral unmixing algorithms that use feedback from measured composite signals and reference spectral data to progressively resolve the contributions of individual dyes. The system compares measured signals against expected spectral patterns and adjusts the decomposition to maximize agreement between observed and predicted signals, thereby maintaining detection accuracy.

Inventive Principle:
Principle #23Feedback

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 method enables the detection of multiple targets in a single assay without the need for additional detection channels, increasing multiplexing capacity while reducing costs and sample volume requirements, thereby enhancing the analytical capabilities of digital assays.

Implementation Method 1

the probe can include a dye that provides a fluorescence signal indicating whether or not the target has been amplified

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Data Source

PatentEP2686449B1Multiplexed digital assays with combinatorial use of signals
Publication Date: 2020.11.18 BIO RAD LABORATORIES INC
  • EP2686449B1 patent drawingFigure 1~2
  • EP2686449B1 patent drawingFigure 3~5
  • EP2686449B1 patent drawingFigure 6~8

AI summary

System, including methods, apparatus, and compositions, for performing a multiplexed digital assay on a greater number of targets through combinatorial use of signals.