Barcode Probe Detection for Multiplexed Nucleic Acid Analysis
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Solution Overview
Problem
Current methods for detecting nucleic acids in biological samples lack accuracy, speed, and sensitivity, particularly in multiplexed detection, identification, and quantification.
Innovation Solution
A method involving probes with a target binding domain and a barcode domain, where the target binding domain recognizes and binds to a specific region of the nucleic acid, and the barcode domain uses multiple attachment regions with distinct sequences for sequential detection of detectable labels, allowing for accurate and sensitive multiplexed detection and quantification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional nucleic acid detection methods are used, then detection can be performed, but accuracy, speed, and sensitivity are insufficient particularly for multiplexed detection
Solution Approach 1:
The probe is divided into distinct functional domains: a target binding domain for specific nucleic acid recognition and a barcode domain with multiple attachment regions for sequential label binding. This segmentation enables simultaneous optimization of detection accuracy through specific binding and detection speed through sequential label association.
Solution Approach 2:
The probe is pre-configured with multiple attachment regions in the barcode domain that can sequentially bind detectable labels. This preliminary structure allows rapid multiplexed detection by enabling sequential association of different labels without requiring separate probe preparations for each target.
2Measurement precision
If conventional detection methods are used, then detection can be performed, but sensitivity is insufficient for multiplexed detection
Solution Approach 1:
A single probe design with multiple attachment regions serves multiple detection functions simultaneously. The barcode domain can sequentially bind different detectable labels to detect multiple target nucleic acids in one assay, providing multi-functionality that enhances sensitivity without proportionally increasing method complexity.
Solution Approach 2:
The barcode domain acts as an intermediary structure between the target binding domain and detectable labels. This mediator enables sequential label binding to multiple attachment regions, amplifying the detection signal and enhancing sensitivity while maintaining a relatively simple overall method framework.
3Adaptability or versatility
If multiple nucleic acids are detected simultaneously, then multiplexed detection is achieved, but the ability to identify specific regions is lost
Solution Approach 1:
Each attachment region in the barcode domain is assigned a distinct sequence that serves as a unique identifier for a specific target region. This local differentiation enables the system to maintain specific region identification capability while detecting multiple nucleic acids simultaneously through sequential label binding to different attachment regions.
Solution Approach 2:
The probe structure adds a temporal dimension to detection by enabling sequential binding of detectable labels to multiple attachment regions. This sequential process in time allows the system to resolve and identify specific target regions even when multiple nucleic acids are detected simultaneously in the same sample.
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 rapid and sensitive detection and identification of multiple nucleic acids by sequentially ordering detectable labels associated with different attachment regions, enhancing the accuracy and efficiency of nucleic acid analysis.
Implementation Method 1
contacting the sample with at least one probe capable of recognizing and binding a first specific region of the at least one target molecule
Implementation Method 2
binding to the first attachment region a first complementary nucleic acid molecule comprising a detectable label
Implementation Method 3
a first complementary nucleic acid molecule of a first reporter complex comprising a detectable label
Data Source
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AI summary
The present invention provides probes, methods, kits, and apparatuses that provide accurate, rapid, and sensitive multiplexed detection, identification, and quantification of target nucleic acids in a sample.