Mask Oligonucleotide Stabilization for Nucleic Acid Detection
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Solution Overview
Problem
Current gene testing methods for detecting bacterial or viral infections and genetic mutations are complex and lack sensitivity in maintaining nucleic acids in a single-stranded state, leading to inaccurate detection and false positives/negatives.
Innovation Solution
The use of mask oligonucleotides to stabilize single-stranded regions in target nucleic acids, allowing for hybridization and detection through nucleic acid chromatography, which enables simple and highly sensitive detection and quantification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If denaturation conditions are eased (neutralized), then the target nucleic acid returns to the double-stranded state, but this prevents probe hybridization to single-stranded regions
Solution Approach 1:
The patent introduces a first oligonucleotide as an intermediary molecule that hybridizes to the target nucleic acid and prevents it from returning to the double-stranded state. This mediator maintains the single-stranded configuration without requiring continuous harsh denaturation conditions, enabling subsequent probe hybridization while simplifying the detection methodology.
2Measurement precision
If high temperature or denaturants are maintained to keep nucleic acid single-stranded, then probe hybridization is enabled, but detection sensitivity decreases due to nonspecific binding
Solution Approach 1:
The patent applies preliminary action by introducing the first oligonucleotide before the detectably labeled oligonucleotide probe. This preliminary oligonucleotide binds to the target nucleic acid and locks it in a single-stranded state, preventing nonspecific binding and secondary structure formation. This preliminary stabilization ensures that subsequent probe hybridization occurs under optimal conditions, enhancing detection accuracy while eliminating the need for harsh denaturation conditions.
3Adaptability or versatility
If secondary structures such as intramolecular loops are present, then nucleic acid maintains natural structure, but probe hybridization is inhibited leading to false negatives
Solution Approach 1:
The first oligonucleotide performs preliminary action by binding to the target nucleic acid and preventing the formation of intramolecular loops and secondary structures. This preliminary stabilization ensures the target remains in an accessible single-stranded state, allowing the detectably labeled oligonucleotide probe to hybridize effectively and accurately detect the target sequence.
4Measurement precision
If complex detection procedures are used to maintain single-stranded state, then detection sensitivity improves, but time consumption and operational complexity increase
Solution Approach 1:
The first oligonucleotide performs preliminary stabilization of the target nucleic acid in a single-stranded state, eliminating the need for time-consuming denaturation steps and continuous harsh condition maintenance. This preliminary action streamlines the detection process, reducing both time consumption and operational complexity while maintaining high detection sensitivity through effective probe hybridization.
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 allows for stable maintenance of single-stranded nucleic acids, enhancing probe hybridization rates and enabling the use of protein labels, resulting in simple, quick, and precise detection of nucleic acids from various organisms.
Implementation Method 1
hybridizing a first oligonucleotide to a target nucleic acid to be detected
Implementation Method 2
a detectably labeled oligonucleotide probe complementary to a portion of the target single-stranded nucleic acid is allowed to hybridize, and then the target nucleic acid is detected using the label as an indicator
Data Source
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AI summary
Very simple, highly sensitive detection or quantification of target nucleic acids of interest has been achieved by: hybridizing mask oligonucleotides to regions in a single-stranded region of a nucleic acid to be assayed between which a region to be hybridized by an oligonucleotide probe is positioned, thereby opening the probe-hybridizing region and keeping the single-stranded region of the target nucleic acid stable, and then subjecting this nucleic acid having the single-stranded region to nucleic acid chromatography.