Raman-Active Genetic Probe for Single Point Variant Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting single point variants in DNA sequences, such as those associated with diseases like Alzheimer's and cancer, face challenges including high costs, time-consuming processes, and limitations due to auto fluorescence and background fluorescence in fluorophore-labelled assays, with a need for improved specificity and sensitivity.
Innovation Solution
The development of genetic probes with Raman-active moieties incorporated into oligonucleotides, allowing for label-free detection of DNA hybridization using Raman spectroscopy, which measures vibrational changes during base pair formation, enabling high specificity and sensitivity at any chosen temperature without generating different read-outs through target strand binding efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorophore-tagged DNA-based probes are used for hybridisation assays, then detection sensitivity is improved, but auto fluorescence and background fluorescence limit the accuracy
Solution Approach 1:
The patent removes the fluorophore tag from the probe system and extracts the detection function to a separate Raman spectroscopy measurement system. This separates the hybridisation probe (which binds to the target) from the detection mechanism (which measures Raman scattering), eliminating the harmful fluorescence background while maintaining sensitivity.
Solution Approach 2:
The patent replaces the optical fluorescence-based detection system with a Raman scattering-based detection system. Instead of using fluorophores that emit light upon excitation, the system uses Raman spectroscopy to detect vibrational scattering, fundamentally changing the detection physics to avoid fluorescence limitations.
2Measurement precision
If enzyme-based methods are used for point variant detection, then accuracy is improved, but the process becomes time-consuming and expensive
Solution Approach 1:
The patent replaces enzyme-based biochemical detection with direct optical detection using Raman spectroscopy. Instead of using enzymes that require incubation time to produce a signal, the system directly measures the Raman scattering from the hybridised DNA probe-target complex, dramatically reducing detection time while maintaining accuracy.
3Reliability
If fluorophore-labelled assays are used, then detection capability is improved, but the system becomes complex and expensive
Solution Approach 1:
The patent extracts the detection function from the probe molecule itself (removing the fluorophore tag) and places it in a separate, dedicated Raman spectroscopy system. This simplifies the probe design to a pure DNA hybridisation probe while using the Raman system as a separate, specialized detection tool.
Solution Approach 2:
The Raman spectroscopy system serves as a universal detection platform that can detect various nucleic acid interactions without requiring probe-specific fluorophore labeling. The system uses the inherent Raman scattering properties of the DNA bases themselves, making the detection method more generally applicable and less complex.
4Device complexity
If Raman spectroscopy is used for label-free detection, then cost and complexity are reduced, but detection sensitivity must be maintained
Solution Approach 1:
The patent optimizes the Raman detection parameters including laser wavelength selection, integration time, and spectral resolution to maximize the sensitivity of the label-free detection. By carefully controlling these parameters, the system achieves sufficient sensitivity for single-point variant detection without requiring fluorescent labels.
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 provides rapid, reliable, and cost-effective detection of single point variants, capable of distinguishing between matched and mismatched bases, improving the diagnostic capabilities for genetic diseases by overcoming the limitations of existing fluorescence-based methods.
Implementation Method 1
Raman spectroscopy measures the light scattering caused by changes in the vibrational properties of the molecule due to bond switching during chemical reaction, conformational changes, and non-bonding (electrostatic, dipolar, hydrogen bonding etc.) interactions.
Data Source
AI summary
The invention relates to a genetic probe, wherein the genetic probe comprises: an oligonucleotide, or an oligonucleotide analogue, with a Raman-active moiety incorporated therein, wherein the Raman-active moiety is incorporated into a base of the oligonucleotide or oligonucleotide thereof; and associated methods, uses, kits and compositions for determining a single point variant nucleotide in a target nucleic acid.