Nucleic Acid Detection at Different Temperatures With One Label
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Solution Overview
Problem
Conventional real-time detection methods for nucleic acid sequences require multiple types of labels or melting analysis to differentiate between multiple sequences, leading to inefficiencies and increased complexity.
Innovation Solution
The method employs different detection temperatures for multiple nucleic acid sequences using a single type of label and detector, allowing for the detection of two or more sequences by adjusting signal generation based on hybridization and cleavage temperatures.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If different types of labels are used to detect multiple target nucleic acid sequences, then detection capability for multiple sequences is improved, but device complexity and cost increase
Solution Approach 1:
The patent changes the detection temperature parameter to differentiate between multiple target sequences. Each probe is designed with a specific melting temperature (Tm) corresponding to its target sequence, allowing detection at different temperatures using the same fluorescent label and detector, thus avoiding the need for multiple labels and reducing device complexity
Solution Approach 2:
The patent makes a single fluorescent label and detector system capable of detecting multiple different target sequences by utilizing temperature differentiation. The same detection system performs multiple detection functions by adjusting the detection temperature to match the Tm of different probes, achieving multi-functionality without increasing device complexity
2Adaptability or versatility
If melting analysis is used to detect multiple target nucleic acid sequences with a single label, then detection capability is improved, but detection time increases
Solution Approach 1:
The patent segments the detection process into multiple temperature stages, where each stage detects a specific target sequence at its optimal temperature. This allows parallel detection of multiple sequences at different temperatures within a single reaction vessel, significantly reducing the time required compared to sequential melting analysis of each sequence individually
3Adaptability or versatility
If probes with different Tm values are designed to detect multiple sequences, then detection capability is improved, but probe design difficulty increases
Solution Approach 1:
The patent systematically adjusts probe parameters (length, composition, GC content) to achieve specific Tm values for different targets. By changing these parameters, probes can be designed to have distinct melting temperatures that correspond to their target sequences, enabling temperature-based differentiation while maintaining ease of design through parameter optimization
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 enhances convenience, cost-effectiveness, and efficiency by enabling the detection of multiple sequences using a single label and detector, reducing the need for multiple labels and simplifying the detection process.
Implementation Method 1
labeled probes or primers specifically hybridized with target nucleic acid sequences
Implementation Method 2
signals generated from fluorescent labels at a selected detection temperature
Implementation Method 3
detection of two target nucleic acid sequences having different detection temperatures
Data Source
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AI summary
The present invention relates to detection of target nucleic acid sequences using different detection temperatures. The present invention employing different detection temperatures enables to detect a plurality of target nucleic acid sequences in conventional real-time manners even with a single type of label in a single reaction vessel. The conventional technologies detect a plurality of target nucleic acid sequences by a melting analysis after target amplification. Unlikely, the present invention does not require a melting analysis after target amplification, such that the time for analysis is greatly reduced.