Isothermal Amplification Components for Rapid Nucleic Acid Testing
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Solution Overview
Problem
Traditional nucleic acid amplification methods, such as PCR, require thermocycling and specialized machinery, making them time-consuming and unsuitable for rapid on-site or point-of-care diagnostics.
Innovation Solution
Isothermal amplification methods using hyperthermophile polymerase and complementary oligonucleotides to amplify nucleic acids without thermocycling, allowing for rapid nucleic acid amplification in a single vessel within 10 minutes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If thermocycling is used for nucleic acid amplification, then amplification can be achieved, but the process becomes time-consuming and requires specialized machinery
Solution Approach 1:
The patent changes the temperature parameter from cyclic (thermocycling) to constant (isothermal), eliminating the need for complex temperature cycling machinery while maintaining amplification efficiency. The reaction is performed at a single temperature of 65°C or higher, simplifying the device requirements.
Solution Approach 2:
The patent extracts and eliminates the thermocycling step from the amplification process, retaining only the essential isothermal extension step. This removes the need for specialized thermocycling machinery while preserving the core amplification function.
2Productivity
If thermocycling is used for nucleic acid amplification, then amplification can be achieved, but the process becomes time-consuming
Solution Approach 1:
The patent implements continuous isothermal extension without interruption by maintaining a constant temperature of 65°C or higher throughout the reaction. This eliminates the time lost during temperature transitions in thermocycling, enabling rapid amplification in less than 10 minutes.
Solution Approach 2:
By changing from cyclic temperature parameters to a constant high temperature parameter, the patent eliminates the time-consuming heating and cooling cycles, achieving rapid amplification in a continuous isothermal process.
3Ease of operation
If standard polymerase is used, then amplification can proceed, but the process requires denaturation steps that complicate the protocol
Solution Approach 1:
The patent changes the operating temperature to 65°C or higher, which allows the polymerase to function without requiring separate denaturation steps. The high constant temperature simultaneously achieves denaturation and extension, simplifying the protocol to a single isothermal step.
Solution Approach 2:
The patent merges the denaturation and extension steps into a single isothermal process by using a temperature (65°C or higher) that accomplishes both functions simultaneously, eliminating the need for separate temperature cycling steps.
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 quick and efficient nucleic acid amplification without specialized equipment, facilitating rapid detection of target sequences in biological samples.
Implementation Method 1
at least one component providing hyperthermophile polymerase activity
Implementation Method 2
at least one oligonucleotide, which at least one oligonucleotide comprises a polynucleotide complementary to a target sequence in the sample nucleic acid
Data Source
AI summary
The technology relates in part to methods and compositions for isothermal amplification of nucleic acids.