L-Nucleotide Tail Capture Probes for Low-Interference Nucleic Acid Capture
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Solution Overview
Problem
Existing nucleic acid capture methods face challenges in achieving high specificity and sensitivity, particularly in the presence of non-target nucleic acids, leading to false positives and compromised data interpretation in diagnostic and analytical techniques.
Innovation Solution
The use of chimeric capture probes with a D-nucleic acid first segment and an L-nucleic acid second segment, which hybridize specifically to target nucleic acids via immobilized L-nucleic acid probes, preventing non-specific binding and enhancing capture specificity and sensitivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional capture probes with standard nucleic acid sequences are used, then the capture process is simple and straightforward, but non-specific binding occurs leading to reduced specificity and false positives
Solution Approach 1:
The capture probe is designed as a composite structure containing both D-nucleic acid segments (for specific target binding) and L-nucleic acid segments (for immobilization). This composite approach allows the probe to simultaneously achieve high specificity through D-nucleic acid hybridization while preventing non-specific binding of natural nucleic acids to the immobilized L-nucleic acid portion, thereby resolving the contradiction between reliability and complexity.
Solution Approach 2:
The L-nucleic acid portion acts as an intermediary that mediates the immobilization of the capture probe to the solid support without interfering with the specific D-nucleic acid to target binding. This intermediary element enables the probe to be固定在支持物上 while maintaining high specificity, as natural D-nucleic acids cannot hybridize to the L-nucleic acid immobilized portion.
2Productivity
If capture probes are immobilized directly on solid support, then the immobilization process is simplified, but hybridization efficiency decreases due to reduced accessibility
Solution Approach 1:
The capture probe is segmented into distinct functional regions: a D-nucleic acid segment for target hybridization and an L-nucleic acid segment for immobilization. This segmentation allows the probe to first hybridize to the target in solution (maintaining high efficiency) and then be immobilized via the L-nucleic acid portion, separating the hybridization and immobilization functions to resolve the contradiction between productivity and ease of operation.
Solution Approach 2:
The capture probe performs preliminary hybridization to the target nucleic acid in solution before being immobilized on the solid support. This preliminary action ensures that the probe maintains full accessibility and hybridization efficiency during the capture step, while the subsequent immobilization of the L-nucleic acid portion provides stable fixation without interfering with the already-formed probe-target complex.
3Measurement precision
If standard nucleic acid sequences are used in capture probes, then the probes are easy to synthesize and handle, but they bind non-specifically to non-target nucleic acids reducing assay accuracy
Solution Approach 1:
The probe uses a composite of D- and L-nucleic acid sequences, where the D-segment provides specific target recognition and the L-segment provides immobilization capability. This composite design enhances detection accuracy by preventing non-specific binding (since natural D-nucleic acids cannot hybridize to immobilized L-nucleic acid) while remaining synthesizable through standard oligonucleotide synthesis methods that can incorporate both enantiomers.
Solution Approach 2:
Different portions of the capture probe have different properties: the D-nucleic acid portion is designed for high-specificity binding to the target sequence, while the L-nucleic acid portion is designed specifically for immobilization and has no affinity for natural nucleic acids. This local differentiation of function within the probe structure achieves high measurement precision without significantly complicating the overall synthesis process.
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 significantly improves the purity and reliability of nucleic acid capture, reducing non-target interference and enhancing the accuracy of subsequent analytical techniques such as sequencing and amplification assays.
Implementation Method 1
the second segment of the capture probe and the immobilized probe comprise homopolymers of L-nucleic acids that can hybridize to one another
Implementation Method 2
the first segment includes a D nucleic acid of at least 10 D-nucleobase units complementary to the target nucleic acid
Data Source
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AI summary
There is disclosed a method of capturing a target nucleic acid, comprising: contacting a target nucleic acid with a capture probe and an immobilized probe, the capture probe comprising a first segment that binds to the target nucleic acid and a second segment that binds to the immobilized probe, wherein the second segment of the capture probe and the immobilized probe comprise L-nucleic acids that can hybridize to one another, wherein the target nucleic acid binds to the first segment of the capture probe, and the second segment of the capture probe binds to the target, thereby capturing the target nucleic acid.