Chain-elongation nucleic acid set for short-chain detection
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Solution Overview
Problem
Current methods face challenges in detecting short-chain nucleic acids, such as siRNA and miRNA, due to their short sequences, which make amplification difficult and reduce sensitivity.
Innovation Solution
A method involving a chain-elongation nucleic acid set that elongates short-chain nucleic acids to form long-chain nucleic acids, using a universal primer set for multiplex amplification and detection with a probe immobilized substrate to detect hybridization, allowing for simultaneous detection of multiple target nucleic acids.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional amplification techniques are used for short-chain nucleic acids, then the detection process is simple, but the sensitivity is low due to insufficient amplification
Solution Approach 1:
The patent applies preliminary action by performing chain elongation on short-chain nucleic acids before amplification. The chain-elongation nucleic acids hybridize to the target short-chain nucleic acids and extend their length, creating longer templates that can be effectively amplified by subsequent PCR using universal primers. This pre-processing step resolves the fundamental problem that short-chain nucleic acids lack sufficient length for effective primer annealing and amplification.
2Productivity
If multiple short-chain nucleic acids are detected using traditional methods, then individual detection is possible, but simultaneous multiplex detection is difficult
Solution Approach 1:
The patent implements universality by designing chain-elongation nucleic acids with standardized functional regions (amplification regions and detection regions) that can be combined with different target-specific regions. This modular design allows a single set of universal amplification primers to amplify multiple different target sequences simultaneously, enabling multiplex detection. The standardized structure of chain-elongation nucleic acids makes them compatible with universal reagents, achieving multi-functionality across different target detection scenarios.
Solution Approach 2:
The patent applies segmentation by dividing the chain-elongation nucleic acid into distinct functional segments: a target hybridization region, an amplification region, and a detection region. This segmentation allows different regions to serve different purposes in the detection process and enables the use of universal primers that bind to conserved amplification regions across multiple targets, facilitating simultaneous amplification of multiple targets in a single reaction.
3Measurement precision
If chain elongation is performed to increase sensitivity, then detection sensitivity improves, but the detection process becomes more complex
Solution Approach 1:
The patent resolves the complexity issue by making the chain-elongation process compatible with universal reagents. The amplification regions of chain-elongation nucleic acids are designed to be recognized by universal amplification primers, and the detection regions are designed to be recognized by universal detection probes. This universality means that the same reagents can be used for chain elongation and subsequent amplification/detection of multiple targets, reducing operational complexity despite the additional chain elongation step.
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 method enhances the sensitivity and accuracy of detecting multiple short-chain nucleic acids simultaneously, overcoming the limitations of traditional amplification techniques by converting short sequences into longer ones for effective detection.
Implementation Method 1
obtaining a target nucleic acid or a complementary sequence thereof and a long-chain nucleic acid group containing a first sub-chain-elongation nucleic acid corresponding thereto and a second sub-chain-elongation nucleic acid corresponding thereto
Implementation Method 2
obtaining an amplification product group by maintaining the long-chain nucleic acid group and the primer set under amplification conditions
Implementation Method 3
detecting presence/absence and/or an amount of hybridization between the amplification product group and a first probe
Data Source
AI summary
According to one embodiment, a detection method is a method for detecting a plurality of target nucleic acids in a sample. The method includes (a) preparing a chain-elongation nucleic acid set group, a primer set, and a probe immobilized substrate, (b) obtaining the target nucleic acid and a long-chain nucleic acid group containing a first sub-chain-elongation nucleic acid and a second sub-chain-elongation nucleic acid, (c) obtaining an amplification product group by maintaining the long-chain nucleic acid group and the primer set under amplification conditions, (d) detecting presence/absence and/or an amount of hybridization, and (e) detecting the plurality of target nucleic acids.