Two-Probe Tm Detection for Multiplex Nucleic Acid Testing
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Conventional real-time detection methods for nucleic acids can only detect a single target nucleic acid per label, limiting the number of nucleic acids that can be simultaneously detected in a single reaction.
Innovation Solution
A method using two probes with different Tm values that hybridize adjacently to the target nucleic acid, allowing for the detection of multiple nucleic acids in real-time using a single type of label by employing signal-changing and signal-constant temperature ranges.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If conventional real-time detection methods using labeled probes or primers are used, then a single target nucleic acid can be detected, but the number of target nucleic acids that can be simultaneously detected is limited by the number of labels (5 or less)
Solution Approach 1:
The invention changes the detection parameter from label-based identification to temperature-based identification. By using two probes with different Tm values (melting temperatures), the system can distinguish between multiple target nucleic acids by detecting signals at different detection temperatures, eliminating the need for multiple labels
Solution Approach 2:
A single type of label is made universal for detecting multiple target nucleic acids. The same label can be used for all targets, and differentiation is achieved through the temperature-dependent hybridization characteristics of the probes rather than requiring different label types or colors
2Quantity of substance
If multiple labels are used to detect multiple target nucleic acids, then the detection capacity increases, but the device complexity and cost increase
Solution Approach 1:
The invention shifts from using multiple labels with different properties to using a single label type with detection differentiated by temperature parameters. The system measures fluorescence signals at multiple detection temperatures corresponding to the Tm values of different probes, simplifying the manufacturing and implementation
3Quantity of substance
If conventional methods with multiple labels are used, then multiple targets can be detected, but the analysis time and cost increase
Solution Approach 1:
The invention merges the detection of multiple target nucleic acids into a single reaction vessel using a single label type. By performing all detections simultaneously at different temperatures in one reaction, the system eliminates the need for separate reactions or sequential analysis, reducing both time and cost
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
Enables the detection of multiple target nucleic acids in a single reaction vessel by adjusting detection temperatures, reducing analysis time and requiring only a single type of label, while maintaining high efficiency and cost-effectiveness.
Implementation Method 1
two probes having different Tm values capable of hybridizing adjacent to the target nucleic acid
Implementation Method 2
detecting signals at n detection temperatures... the composition for detecting the ith target nucleic acid has a signal-changing temperature range
Data Source
Figure 1(i)~1(iii)
Figure 2(i)~2(iii)
Figure 3(i)~3(iii)
AI summary
The present disclosure relates to a method for detecting n target nucleic acids in a sample using a composition for detecting a target nucleic acid, which comprises two probes having different Tm values. The method according to the present disclosure enables the real-time detection of multiple target nucleic acids using a single type of label.