Catalytic Probe Detection of Nucleic Acids Without PCR
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Solution Overview
Problem
Current methods for detecting infectious agents like MRSA and viruses are slow, require specialized skills, and are energy-intensive, relying heavily on PCR which is sensitive to contaminants and costly due to repeated heating and cooling cycles.
Innovation Solution
A method using a probe with a catalytic element and binding element that exposes nucleic acids or proteins to a substrate, causing a physical change, such as precipitation or fluorescence, which entraps identifiable components, allowing for rapid detection without the need for PCR or pre-binding capture steps.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If PCR is used to amplify target oligonucleotide, then detection sensitivity is improved, but detection time and energy consumption increase
Solution Approach 1:
The invention extracts and eliminates the PCR amplification step from the detection process. By using a probe with catalytic element that directly binds to target nucleic acids and produces a measurable signal through substrate reaction, the method removes the time-consuming amplification cycles while maintaining detection sensitivity through direct binding and catalytic signal generation.
Solution Approach 2:
The invention replaces the mechanical/thermal cycling system of PCR with a chemical catalytic system. Instead of repeated heating and cooling cycles to amplify DNA, the method uses a catalytic element on the probe that chemically transforms substrate molecules to produce a detectable signal, eliminating the need for thermal cyclers and reducing both time and energy requirements.
2Measurement precision
If PCR is used to amplify target oligonucleotide, then detection sensitivity is improved, but device complexity and cost increase
Solution Approach 1:
The invention removes the complex PCR system including thermal cyclers, multiple reagent steps, and amplification cycles from the detection device. The simplified system uses a probe with catalytic element that directly detects target nucleic acids through binding and catalytic substrate transformation, eliminating the need for complex instrumentation while maintaining sensitivity.
Solution Approach 2:
The invention replaces the complex mechanical thermal cycling system with a simpler chemical catalytic reaction system. The catalytic element on the probe performs substrate transformation at constant temperature, eliminating the need for thermal cyclers, multiple heating/cooling stages, and complex reaction monitoring systems required by PCR.
3Quantity of substance
If repeated heating and cooling cycles are used in PCR, then target amplification is achieved, but energy consumption increases
Solution Approach 1:
The invention extracts and eliminates the energy-intensive heating and cooling cycles from the detection process. By using a probe with catalytic element that operates at constant temperature through chemical catalysis, the method achieves target detection without the repeated thermal cycling that consumes significant energy in PCR systems.
Solution Approach 2:
The invention replaces the energy-consuming thermal cycling mechanism with a chemical catalytic mechanism. The catalytic element on the probe facilitates substrate transformation at constant temperature, eliminating the need for continuous heating and cooling cycles that drive PCR amplification and consume substantial energy.
4Measurement precision
If PCR is used for detection, then target detection is achieved, but sensitivity to contaminants increases
Solution Approach 1:
The invention removes the PCR amplification step that is highly sensitive to contaminants. By using direct binding of the catalytic probe to target nucleic acids followed by substrate reaction, the method eliminates the amplification process that amplifies contamination effects, providing more robust detection in the presence of contaminants.
Solution Approach 2:
The invention replaces the contamination-sensitive PCR system with a catalytic detection system. The catalytic element on the probe performs substrate transformation based on direct target binding rather than exponential amplification, making the detection less susceptible to contamination since there is no amplification step to propagate contaminant signals.
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 method provides a rapid, reliable, and economical means of detecting target nucleic acids or proteins, reducing detection time and energy consumption while eliminating the need for complex laboratory setups.
Implementation Method 1
The probe also comprises a catalyst (or precursor(s) thereof). The immobilised oligonucleotides are then washed with a mixture of substrate for the catalyst, a second enzyme/substrate pair, matched to the probe system
Implementation Method 2
exposing nucleic acid sequences from a sample to a probe under hybridisation conditions the probe comprising a first catalytic element or precursor thereof and a binding element capable of binding at least partially to the target nucleic acid sequence
Implementation Method 3
The reaction of this second system will cause a measurable shift in the physical state of the system, such as a precipitation, polymerisation, colour change or a change in fluorescence, the formation of which will entrap or otherwise affect the identifiable components
Data Source
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AI summary
A method of testing for the presence of a preselected target nucleic acid, protein or antigen in a biological sample by exposing nucleic acids, proteins or antigens to a probe having a catalytic element and binding element. The catalytic element catalyses at least one reaction that results in a physical change such that identifiable elements provide an indication of the presence of the target.