Surface Plasmon Resonance PCR Thermal Cycling
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Solution Overview
Problem
Current gene sequencing and nucleic acid analysis methods, such as PCR, are limited by slow speed and low sensitivity, which hampers the diagnosis and therapeutic development for genetic diseases, and existing SPR-based DNA detection methods lack simultaneous amplification and real-time detection capabilities.
Innovation Solution
The method involves immobilizing polymerase or primers on a gold surface and using optical stimulation of surface plasmon resonance for rapid thermal cycling, coupled with SPR for real-time detection of PCR amplicons, allowing for simultaneous amplification and detection of nucleic acids at picomolar levels without the need for fluorescent probes or dyes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If conventional PCR methods are used, then nucleic acid amplification can be achieved, but the speed is slow and sensitivity is low
Solution Approach 1:
The patent replaces conventional thermal cycling mechanisms with optical stimulation of surface plasmon resonance. Light energy excites surface plasmons on gold nanoparticles, generating localized heat that drives thermal cycling reactions directly on the sensor surface, eliminating the need for external thermal cyclers and enabling rapid amplification.
Solution Approach 2:
The patent changes the thermal cycling parameters by using optical excitation instead of conventional heating. Surface plasmon resonance converts optical energy to thermal energy at the nanoscale, creating localized temperature changes that enable rapid denaturation, annealing, and extension steps without requiring prolonged heating cycles.
2Measurement precision
If conventional PCR methods are used, then nucleic acid amplification can be achieved, but real-time detection capability is lacking
Solution Approach 1:
The patent merges the amplification and detection functions into a single integrated system. Gold nanoparticles serve dual purposes: catalyzing the PCR amplification reaction through surface plasmon resonance while simultaneously providing the detection signal through their optical properties. This eliminates the need for separate detection steps and enables real-time monitoring.
Solution Approach 2:
The system performs self-detection through the inherent optical properties of the gold nanoparticles. The surface plasmon resonance signal provides direct detection of nucleic acid binding and amplification products without requiring external fluorescent probes or dyes. The amplification process itself generates the detection signal.
3Measurement precision
If fluorescent probes or dyes are used for detection, then detection sensitivity can be improved, but contamination risks increase
Solution Approach 1:
The patent extracts and eliminates the need for fluorescent probes and dyes from the detection system. By using the inherent optical properties of gold nanoparticles and surface plasmon resonance, the system achieves detection without requiring external fluorescent labels that could contaminate the sample or require complex purification steps.
Solution Approach 2:
The system uses disposable gold nanoparticle surfaces that can be easily discarded after use. The nanoparticles remain on the sensor surface during the experiment and can be removed by simple washing, eliminating the need for expensive fluorescent probes that require careful handling and purification to prevent contamination.
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 enables rapid and sensitive nucleic acid amplification and detection, overcoming the limitations of conventional PCR methods by achieving millisecond thermal cycling and picomolar detection sensitivity, suitable for high-throughput analysis and reducing contamination risks.
Implementation Method 1
inducing heating and/or thermal cycling by optical stimulation of surface plasmon resonance resulting in rapid thermal dissipation
Implementation Method 2
immobilizing a polymerase or forward/reverse primers on a gold surface
Implementation Method 3
measuring rates and magnitudes of interaction between the template and molecules immobilized on the gold surface by analyzing mass-induced changes at resonance absorption with SPR
Data Source
AI summary
Methods of performing PCR are provided. Methods may include using an optical source to provide heating for thermocyling the PCR reaction. Methods may include using surface plasmon resonance and/or fluorescence resonance enhanced transfer to allow real-time monitoring of a PCR reaction. Methods may include immobilizing a template, primer, or polymerase on a surface such as a gold or other surface plasmon resonance active surface.