Multiplex Nucleic Acid Amplification with CMOS Sensor Array
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Solution Overview
Problem
Current multiplexed PCR methods face challenges in simultaneously detecting multiple nucleic acid sequences in a single reaction chamber, limiting their ability to efficiently amplify and quantify multiple targets, and require complex fluidic handling procedures.
Innovation Solution
A method and system that utilize a reaction mixture with a primer pair and a sensor array with immobilized probes to detect nucleic acid amplification products in real-time, allowing for the simultaneous amplification and detection of multiple nucleic acid sequences in a single reaction chamber using a CMOS-based detection system.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If multiplexed PCR is used to amplify multiple nucleic acid sequences simultaneously, then productivity is improved, but measurement precision deteriorates due to difficulty in detecting multiple amplicons simultaneously
Solution Approach 1:
The detection system is segmented into multiple individually addressable locations on a solid support, with each location containing probes specific to different target sequences. This allows simultaneous detection of multiple amplicons generated during multiplexed PCR by spatially separating detection events across different locations on the array.
Solution Approach 2:
Solid support-bound probes act as intermediaries between the amplified nucleic acid sequences and the detection system. These probes specifically hybridize to target sequences and generate detectable signals, enabling precise measurement of multiple amplicons simultaneously without cross-interference.
2Measurement precision
If multiple single-plex reactions are run separately, then measurement precision is maintained, but device complexity increases due to complex fluidic handling procedures
Solution Approach 1:
Multiple single-plex reactions are merged into a single multiplexed reaction chamber, eliminating the need for complex fluidic handling between separate reactions. The solid support array enables simultaneous detection of all targets within this single reaction volume, maintaining measurement precision while dramatically simplifying the overall system architecture.
Solution Approach 2:
The solid support array serves multiple functions: it provides spatially separated detection locations for multiple targets, enables simultaneous monitoring of all amplification reactions, and allows for scalable expansion to detect additional targets without increasing fluidic complexity.
3Productivity
If a large number of amplicons are multiplexed, then productivity is improved, but ease of operation deteriorates due to difficulty in simultaneous detection
Solution Approach 1:
The detection system transitions from temporal or sequential detection to spatial detection across a two-dimensional array. Each addressable location on the solid support provides a unique spatial coordinate for detecting specific amplicons, enabling simultaneous monitoring of numerous targets through spatial resolution rather than temporal sequencing.
Solution Approach 2:
The system utilizes detectable signals that change state upon hybridization events, analogous to color changes in traditional assays. Each probe location generates a distinct detectable signal when its target is present, allowing operational simplicity through direct signal readout without complex processing or differentiation procedures.
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 high-sensitivity and real-time monitoring of nucleic acid amplification, allowing for the accurate quantification of multiple targets with improved efficiency and reduced complexity in fluidic handling, achieving high sensitivity and specificity in nucleic acid detection.
Implementation Method 1
probes are capable of capturing the target nucleic acid molecule
Implementation Method 2
an array of detectors configured to detect at least one signal from the addressable locations, wherein the at least one signal is indicative of the presence or absence of the target nucleic acid molecule
Data Source
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AI summary
The present disclosure provides methods, devices and systems that enable simultaneous multiplexing amplification reaction and real-time detection in a single reaction chamber.