Multiplexed Nucleic Acid Detection Using Multifunctional ID Beads
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Solution Overview
Problem
Current methods for large-scale serological testing, such as RNA detection and protein analysis, are hindered by cumbersome purification protocols, enzymatic reactions, and high reagent costs, limiting high-throughput detection capabilities.
Innovation Solution
The method involves using multifunctional ID beads with capture reagents and oligonucleotide handles for simultaneous detection of multiple analytes, enabling unique barcoding and visualization of target analytes and reagents, allowing for large-scale serological testing without the need for extensive purification or enzymatic steps.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If current methods for RNA detection and protein analysis are used, then detection accuracy is maintained, but the process requires cumbersome purification protocols and enzymatic reactions, increasing device complexity and processing time
Solution Approach 1:
The invention extracts and eliminates the cumbersome purification and enzymatic reaction steps from the detection process. By using affinity reagents that directly bind target analytes and ID reagents with capture reagents and barcodes, the method removes the need for complex purification protocols and enzymatic amplification steps, thereby simplifying the overall detection system while maintaining high throughput capability
Solution Approach 2:
The invention employs multifunctional ID reagents that combine capture reagents, barcodes, and optional amplification sequences into a single universal platform. This multi-functional design allows the same basic system to detect various target analytes (proteins, antibodies, nucleic acids, small molecules) without requiring separate complex purification and detection protocols for each analyte type, thus improving throughput while reducing overall system complexity
2Productivity
If current methods with labeled antibodies and enzymatic reactions are used, then target analyte detection is achieved, but reagent cost and processing time increase
Solution Approach 1:
The invention performs preliminary action by pre-attaching barcodes and capture reagents to ID reagents before the detection process. This pre-preparation eliminates the need for time-consuming enzymatic reactions and label attachment steps during actual detection, allowing for rapid processing of large numbers of samples while reducing turnaround time
Solution Approach 2:
The invention uses barcode sequences as information copies that can be read without requiring expensive labeled antibodies or complex enzymatic reactions. The barcode serves as a stable, inexpensive information carrier that replaces costly detection reagents, thereby reducing both reagent costs and processing time while maintaining detection capability
3Adaptability or versatility
If multifunctional ID reagents with barcodes are used, then large scale multiplexed detection is enabled, but reagent design complexity increases
Solution Approach 1:
The invention segments the detection system into distinct functional modules: ID reagents with barcodes, affinity reagents for target capture, and readout systems. This segmentation allows each component to be independently optimized and standardized, reducing overall reagent design complexity while enabling flexible multiplexing configurations for detecting multiple analytes simultaneously across large sample scales
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 rapid, accurate detection of up to 10,000 samples in a single run, reducing costs and turnaround time while maintaining sensitivity and specificity, and allows for multiplexing of RNA and protein analysis.
Implementation Method 1
one or more capture reagents capable of binding to the one or more target analytes or one or more primary affinity regents
Implementation Method 2
one or more ID barcodes; and one or more oligonucleotide handles, wherein one of the oligonucleotide handles identifies a specific primary affinity reagent
Data Source
AI summary
This disclosure herein sets forth embodiments to provide a serological test to detect target analytes that can scale to up to 10,000 or more samples in a single run. This disclosure herein sets forth methods to allow for unique barcoding by using a multi-level barcode scheme that is modular and enables easy detection of multiple analytes in samples.