Nucleic Acid Amplification Kinetics for Internal Control
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Solution Overview
Problem
Current nucleic acid amplification techniques require sophisticated and expensive hardware to differentiate between signals from target and internal control nucleic acids in the same reaction vessel, often necessitating the use of multiple reporter systems and additional detection steps, which increases complexity and costs.
Innovation Solution
The method involves conducting nucleic acid amplification reactions with different kinetics for the target and internal control in the same reaction vessel, allowing for signal differentiation through varying amplification kinetics, such as exponential vs. non-exponential processes, without the need for multiple reporter systems, using algorithms to analyze signal readings against time to distinguish between the two amplification processes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple reporter systems are used to differentiate between target and internal control signals, then measurement precision is improved, but device complexity and cost increase
Solution Approach 1:
The patent applies parameter changes by utilizing different amplification kinetics (exponential vs non-exponential) as a distinguishing parameter between target and internal control signals. Instead of using multiple reporter systems with different emission wavelengths, the invention changes the kinetic parameters of the amplification reactions themselves, allowing differentiation through time-dependent signal analysis with a single reporter system.
Solution Approach 2:
The patent uses algorithms to create a computational copy or model of the expected signal patterns from exponential and non-exponential amplification processes. By comparing the actual detected signal against these predefined kinetic patterns, the system can differentiate between target and control signals without requiring additional physical detection hardware.
2Measurement precision
If multiple reporter systems are used to differentiate between target and internal control signals, then measurement precision is improved, but cost increases
Solution Approach 1:
The invention changes the kinetic parameters of amplification reactions to create distinguishable signal patterns, eliminating the need for expensive multi-channel detection hardware. This parameter-based differentiation approach significantly reduces system cost while maintaining the ability to distinguish between target and internal control signals.
3Measurement precision
If additional detection steps are implemented to differentiate signals, then measurement precision is improved, but productivity decreases
Solution Approach 1:
The patent maintains continuous signal detection throughout the amplification process, analyzing kinetic patterns in real-time without interrupting the reaction. This continuous monitoring approach allows for immediate differentiation between target and control signals, eliminating the need for separate detection steps and preserving high productivity.
Solution Approach 2:
The invention replaces mechanical or procedural separation steps with computational analysis of temporal signal patterns. By using algorithms to distinguish exponential from non-exponential kinetics during the ongoing reaction, the system eliminates additional manual detection steps while maintaining accurate signal differentiation.
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 simplifies the hardware requirements by enabling effective internal control monitoring without the need for expensive multi-signal detection systems, reducing costs and complexity while maintaining accurate differentiation between target and internal control signals.
Implementation Method 1
nucleic acid amplification in which product is detectable by the presence of a signal generated by polynucleic acid formation from the first polynucleotide
Data Source
AI summary
A method for determining the presence and/or amount of a first polynucleic acid in a sample comprising subjecting the sample to nucleic acid amplification in which product is detectable by the presence of a signal generated by polynucleic acid formation from the first polynucleotide characterised in that the nucleic acid amplification reaction is conducted, in the same reaction vessel, with a predetermined amount of a second polynucleic acid which is subjected to nucleic acid amplification, the product of which is detectable by the presence of the same signal generated by polynucleic acid formation from the second polynucleotide as that generated by polynucleic acid formation from the first polynucleotide and wherein the product of the second polynucleic acid is produced with different reaction kinetics from the product of the first polynucleic acid such that the second polynucleic acid acts as an internal control for the method.