Multiplexed Biochemical Assay Signal Encoding via Probe Segmentation
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Solution Overview
Problem
Current multiplexing technologies relying on fully non-degenerate coding schemes are limited by reagent concentration requirements and signal detection capabilities, making them inefficient for detecting multiple analytes in a single signal measurement.
Innovation Solution
The method involves contacting a sample with multiple subsets of hybridization probes to generate cumulative signal measurements, which are then compared to uniquely identify combinations of polynucleotide analytes, allowing for unambiguous detection without the need for fully non-degenerate coding schemes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fully non-degenerate coding schemes are used to detect multiple analytes in a single signal measurement, then measurement precision is improved, but device complexity and reagent concentration requirements increase
Solution Approach 1:
The patent divides the sample into multiple subsets and contacts each subset with different hybridization probes to generate separate cumulative signal measurements. This segmentation allows the system to detect multiple analytes without requiring fully non-degenerate coding schemes, as each subset provides partial information that collectively resolves analyte identities.
Solution Approach 2:
The patent transitions from a single-dimensional signal measurement approach to a multi-dimensional approach by comparing cumulative signal measurements across multiple subsets. This dimensional expansion enables unambiguous analyte detection through pattern recognition across multiple measurement dimensions rather than relying on complex coding within a single measurement.
2Measurement precision
If fully non-degenerate coding schemes are used to detect multiple analytes, then measurement precision is improved, but reagent concentration requirements increase
Solution Approach 1:
By segmenting the detection process into multiple subsets with different probe combinations, the system reduces the reagent concentration requirements for each individual probe while maintaining overall detection precision. The cumulative signal measurements from multiple subsets compensate for lower individual reagent concentrations.
3Adaptability or versatility
If multiple subsets of hybridization probes are used to detect multiple analytes, then analyte detection capability is improved, but device complexity increases
Solution Approach 1:
The patent segments the analyte detection process into multiple subsets, where each subset contacts different hybridization probes with the sample. This segmentation enables the detection of multiple analytes simultaneously while managing complexity through modular probe design and systematic comparison of cumulative signal measurements.
4Measurement precision
If cumulative signal measurements from multiple subsets are compared, then measurement precision is improved, but loss of time increases
Solution Approach 1:
The patent performs preliminary actions by contacting multiple subsets of the sample with different hybridization probes in parallel or sequential fashion before final analyte identification. This preliminary measurement of cumulative signals from multiple subsets enables accurate analyte identification while minimizing total detection time through efficient experimental design.
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 enables the unambiguous detection of multiple analytes in a single optical channel, improving the efficiency and reducing the complexity of reagent concentrations and signal detection requirements.
Implementation Method 1
contacting a first subset of the sample with a first plurality of hybridization probes to generate a first cumulative signal measurement comprising one or more signals generated from the first plurality of hybridization probes
Implementation Method 2
each hybridization probe of the first plurality of hybridization probes and the one or more additional plurality of hybridization probes comprises a fluorophore capable of being detected in the same optical channel
Data Source
AI summary
The present disclosure provides methods and compositions for the multiplexed detection of multiple analytes from a sample. Analytes may be nucleic acid analytes. Detection of analytes may comprise contacting one or more sample subsets with hybridization probes, thereby generating one or more cumulative signal measurements capable of detecting the presence of absence of a plurality of analytes.