Nucleic Acid Detection via Photocrosslinking Stabilization
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Solution Overview
Problem
Existing nucleic acid detection methods face challenges in accurately detecting target nucleic acid molecules due to interference from non-specific association products and the solid phase carrier, which affects the accuracy and reliability of the detection process.
Innovation Solution
A method involving a sample solution with a first nucleic acid probe labeled with a luminescent substance and a second nucleic acid probe that specifically hybridizes with the target nucleic acid molecule, forming covalent bonds through a photocrosslinking reaction, followed by binding to a solid phase carrier and releasing the luminescent marker for detection, thereby stabilizing the association product and reducing non-specific interactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a labeled probe is used to detect target nucleic acid molecules through hybridization, then detection capability is provided, but non-specific association products are formed leading to reduced measurement precision
Solution Approach 1:
The probe is pre-modified with a photoreactive group before hybridization. After specific hybridization with the target nucleic acid, UV irradiation is applied to form covalent bonds between the probe and target, stabilizing the association product and preventing non-specific binding. This preliminary modification enables subsequent selective stabilization that eliminates harmful non-specific association products.
Solution Approach 2:
The patent replaces traditional mechanical separation methods with a photochemical mechanism. UV irradiation induces photocrosslinking that forms covalent bonds, substituting physical separation techniques with a chemical bonding approach to achieve more effective elimination of non-specific association products.
2Reliability
If the association product is stabilized by covalent bonding, then non-specific association is suppressed, but the complexity of the detection procedure increases
Solution Approach 1:
The photoreactive group is incorporated into the probe structure, allowing continuous UV irradiation during the hybridization process to simultaneously achieve specific binding and covalent stabilization in one operation. This eliminates the need for separate stabilization steps, maintaining procedural simplicity while ensuring reliable specific association.
3Measurement precision
If the luminescent marker is separated from the solid phase carrier, then detection sensitivity is improved, but the procedure requires additional separation steps
Solution Approach 1:
The detection system is segmented into distinct functional components: the luminescent marker attached to the probe, the target nucleic acid, and the solid phase carrier. After covalent stabilization, the luminescent marker remains with the probe-target complex, allowing selective separation from the carrier through simple filtration or centrifugation, achieving sensitive detection without time-consuming 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
This approach enables high-accuracy detection of target nucleic acid molecules by stabilizing the association product and separating the luminescent marker from the solid phase carrier, minimizing interference and improving detection sensitivity.
Implementation Method 1
a first nucleic acid probe that has a luminescent substance in the form of a first marker bound thereto and specifically hybridizes with the target nucleic acid molecule, and a second nucleic acid probe that has a second marker bound thereto and specifically hybridizes with a target nucleic acid molecule
Implementation Method 2
forming covalent bonds through a photocrosslinking reaction
Data Source
AI summary
A method for detecting a target nucleic acid comprising: forming a three-component association product by allowing the association of at least a nucleic acid molecule, a first nucleic acid probe having a first marker bound thereto, and a second nucleic acid probe having a second marker bound thereto; forming at least one covalent bond between the target nucleic acid molecule and the first nucleic acid probe and between the target nucleic acid molecule and the second nucleic acid probe; and binding the three-component association product to a solid phase carrier through the second marker; recovering the three-component association product bound to the solid phase carrier; releasing the first marker from the recovered three-component association product; and detecting the target nucleic acid molecule by detecting the free first marker.