Blocker Nucleic Acid for RNA Detection
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Solution Overview
Problem
Existing RNA detection methods face challenges in achieving high sensitivity and accuracy, particularly in detecting short RNA sequences and preventing non-specific reactions.
Innovation Solution
The method involves using a sensor DNA with a target RNA recognition site and a module region, along with a blocker nucleic acid that complements the module region of the sensor DNA, to inhibit non-specific amplification during PCR.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional RNA detection methods are used, then detection can be performed, but sensitivity and accuracy are insufficient especially for short RNA sequences
Solution Approach 1:
A sensor DNA molecule is introduced as an intermediary that contains both a target RNA recognition site and a module region. The sensor DNA hybridizes with target RNA and serves as a template for polymerization, enabling detection of short RNA sequences with high sensitivity and accuracy without requiring complex detection systems.
Solution Approach 2:
The detection method utilizes changes in molecular parameters during polymerization - when sensor DNA hybridizes with target RNA, the polymerase can extend the sensor DNA using the RNA as template. This parameter change (polymerization occurrence) provides a measurable signal that indicates target RNA presence with high precision.
2Measurement precision
If sensor DNA is used for RNA detection, then detection capability is improved, but non-specific reactions and false positives increase
Solution Approach 1:
A blocker nucleic acid is introduced that is complementary to the module region of the sensor DNA. This blocker preliminarily prevents non-specific amplification of free sensor DNA by hybridizing with it, thereby eliminating false positive signals before they can interfere with the detection results.
Solution Approach 2:
The potential harm of non-specific sensor DNA amplification is converted into a benefit by using the blocker nucleic acid to specifically suppress only the non-specific signals. The blocker itself becomes a useful reagent that enhances detection accuracy by eliminating false positives while leaving specific target detection unaffected.
3Reliability
If detection sensitivity is increased to detect short RNA sequences, then early diagnosis capability is improved, but false positive rates increase due to non-specific reactions
Solution Approach 1:
The sensor DNA acts as an intermediary that provides high sensitivity detection through its polymerization capability, while the blocker nucleic acid acts as a second intermediary that ensures signal specificity by suppressing non-specific reactions. Together, they enable reliable early diagnosis with high confidence.
Solution Approach 2:
The sensor DNA is designed with different functional regions having different properties: the target recognition site provides high affinity binding for sensitivity, while the module region serves as a specific target for the blocker to ensure specificity. This local differentiation of function allows simultaneous achievement of sensitivity and specificity.
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 enhances the sensitivity and accuracy of RNA detection, allowing for the analysis of short RNA sequences with high precision and low detection limits at the femtomole and attomole levels.
Implementation Method 1
a) hybridizing a sensor DNA, which includes a sequence complementary to a target RNA to be detected, with the target RNA
Implementation Method 2
b) performing polymerization using a polymerase, in which a module region of the sensor DNA is used as a template and the target RNA is used as a primer
Implementation Method 3
c) treating the blocker nucleic acid that complementarily binds to the sensor DNA thereto to inhibit amplification of the sensor DNA that does not hybridize with target RNA
Data Source
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AI summary
The present invention relates to a method for detecting a target RNA, which includes treating a blocker nucleic acid. In particular, the method of the present invention can inhibit non-specific reactions through the treatment of the blocker nucleic acid, and analyze even short RNA sequences, thereby enabling detection with high sensitivity and accuracy, and thus can be widely utilized for the diagnosis of various diseases such as infections and cancers.