Nucleic Acid Detection via Multi-Probe Hybridization
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Solution Overview
Problem
Current nucleic acid detection methods, such as sandwich hybridization, require nucleic acid amplification techniques like PCR, which can lead to false positivity due to amplified contaminants, and are labor-intensive for multi-gene detection, while techniques without amplification often require special enzymes or equipment.
Innovation Solution
A method involving simultaneous hybridization of multiple detection probes with different regions of the target nucleic acid, without amplification or sensitization, to achieve high sensitivity detection using a capture probe immobilized on a support.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If nucleic acid amplification techniques like PCR are used to increase detection sensitivity, then detection sensitivity is improved, but false positivity occurs due to amplified contaminants
Solution Approach 1:
The invention uses multiple detection probes that bind to different regions of the target nucleic acid sequence. By segmenting the detection process across multiple probe binding sites rather than relying on a single amplification event, the method achieves high sensitivity without the need for PCR amplification, thereby avoiding false positivity from amplified contaminants.
Solution Approach 2:
The invention introduces a capture probe as an intermediary element that immobilizes the target nucleic acid on a support surface. This capture probe acts as a mediator between the target nucleic acid and the detection probes, enabling sensitive detection through the formation of a stable sandwich structure without requiring nucleic acid amplification.
2Adaptability or versatility
If multiple primer sets are used to detect multiple genes simultaneously, then multi-gene detection capability is improved, but quality control of primer sets requires much labor
Solution Approach 1:
The invention employs a universal capture probe design that can immobilize various target nucleic acids on the support. By using the same capture probe infrastructure for different genes and simply changing the detection probes that recognize specific gene sequences, the system achieves multi-gene detection capability without requiring separate primer sets for each gene, thereby simplifying quality control.
3Measurement precision
If sensitization techniques with labeling substances or tag sequences are used to achieve high sensitivity without amplification, then detection sensitivity is improved, but special enzymes or complex reactions are required
Solution Approach 1:
The invention extracts and eliminates the need for complex sensitization techniques, special enzymes, and tag sequences by using a simpler approach: multiple detection probes that directly hybridize to different regions of the target nucleic acid. The signal detection is achieved through the binding itself rather than through additional enzymatic reactions or labeling steps, thereby reducing device complexity while maintaining high sensitivity.
4Measurement precision
If sensitization techniques with special support or optical planar waveguide are used to achieve high sensitivity without amplification, then detection sensitivity is improved, but special equipment is required
Solution Approach 1:
The invention replaces expensive, specialized equipment like optical planar waveguides with simple, disposable support surfaces such as microtiter plates or slide glasses. The capture probes are immobilized on these inexpensive supports, and the detection is performed using conventional detection methods, thereby achieving high sensitivity without requiring special equipment while maintaining ease of use and low cost.
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 sensitive detection of nucleic acids without the need for amplification or special equipment, reducing false positivity and labor requirements, and allowing for simultaneous detection of multiple genes.
Implementation Method 1
bringing a target nucleic acid or fragmentation product thereof, a plurality of detection probes, and a capture probe immobilized on a support, into contact with each other to hybridize the capture probe with the target nucleic acid or fragmentation product thereof
Implementation Method 2
to hybridize the target nucleic acid or fragmentation product thereof with the plurality of detection probes, thereby binding the plurality of detection probes to the support through the capture probe and the target nucleic acid or fragmentation product thereof
Data Source
Figure 1A~1C
Figure 2A~2C
Figure 3
AI summary
Disclosed is a method for detecting nucleic acid with high sensitivity even in cases where the target nucleic acid is detected by sandwich hybridization using neither nucleic acid amplification nor a sensitization technique. The method for detecting a target nucleic acid comprises the steps of: sequentially or simultaneously bringing a target nucleic acid or fragmentation product thereof, a plurality of detection probes, and a capture probe immobilized on a support, into contact with each other to hybridize the capture probe with the target nucleic acid or fragmentation product thereof and to hybridize the target nucleic acid or fragmentation product thereof with the plurality of detection probes, thereby binding the plurality of detection probes to the support through the capture probe and the target nucleic acid or fragmentation product thereof; and then detecting the plurality of detection probes bound to the support.