Real-Time Microarray System for Multiplexed Nucleic Acid Detection
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Solution Overview
Problem
Current microarray technologies face challenges in achieving high-quality measurements of multiple analytes simultaneously due to inherent noise, systematic errors, and unpredictable incubation times, which limit the dynamic range and accuracy of nucleic acid sequence amplification and detection, especially in multiplexed Q-PCR and microarray systems.
Innovation Solution
A real-time microarray system that measures binding of analytes to probes on a solid substrate at multiple time points, using fluorescent or luminescent signals, allowing for the determination of analyte concentration without the need for washing or saturation, and incorporating quenching moieties to enhance signal detection and reduce background noise.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional fluorescent-based microarrays are used to detect captured analytes after incubation, then the detection process can be completed, but the signal measurements are compromised by floating labeled species and washing artifacts
Solution Approach 1:
The patent replaces conventional fluorescent-based optical detection with surface plasmon resonance (SPR) detection, which is a label-free, real-time detection method. This substitution eliminates the need for fluorescent labels and washing steps, thereby removing the harmful interference from floating labeled species and washing artifacts while maintaining measurement precision.
Solution Approach 2:
The patent implements continuous real-time detection during the incubation process using SPR, rather than discontinuous end-point detection after incubation. This continuous monitoring allows for accurate measurement of binding events as they occur, eliminating the need for washing steps and avoiding washing artifacts that compromise signal measurement accuracy.
2Reliability
If incubation is performed to allow analyte-probe binding, then binding occurs, but washing artifacts occur when solution is removed
Solution Approach 1:
The patent replaces fluorescent label-based detection with label-free surface plasmon resonance detection, which enables real-time monitoring of binding events without requiring washing steps. This substitution eliminates washing artifacts while maintaining reliable binding detection, as the SPR signal continuously reflects the mass changes on the sensor surface during incubation.
3Productivity
If multiple analytes are detected simultaneously in conventional microarrays, then high throughput is achieved, but dynamic range and accuracy are limited
Solution Approach 1:
The patent replaces conventional fluorescent microarray detection with surface plasmon resonance detection, which provides real-time, label-free measurement of binding events. This substitution enables simultaneous detection of multiple analytes with enhanced dynamic range and accuracy, as SPR can quantify binding in real-time without the signal saturation and background interference problems that limit conventional fluorescent methods.
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 accurate and sensitive determination of binding characteristics and analyte concentrations, improving the dynamic range and reducing systematic errors, allowing for the simultaneous measurement of multiple nucleic acid sequences in a single amplification reaction without the need for sample splitting or extensive washing.
Implementation Method 1
using fluorescent or luminescent signals
Implementation Method 2
incorporating quenching moieties to enhance signal detection and reduce background noise
Data Source
AI summary
This present disclosure provides methods and systems for measuring the concentration of multiple nucleic acid sequences in a sample. The nucleic acid sequences in the sample are simultaneously amplified, for example, using polymerase chain reaction (PCR) in the presence of an array of nucleic acid probes. The amount of amplicon corresponding to the multiple nucleic acid sequences can be measured in real-time during or after each cycle using a real-time microarray. The measured amount of amplicon produced can be used to determine the original amount of the nucleic acid sequences in the sample. Also provided herein are biosensor arrays, systems and methods for affinity based assays that are able to simultaneously obtain high quality measurements of the binding characteristics of multiple analytes, and that are able to determine the amounts of those analytes in solution. The present disclosure also provides a fully integrated bioarray for detecting real-time characteristics of affinity based assays.