Virus Detection via Antibody Capture and Fluorescence Microscopy
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Solution Overview
Problem
Current methods for detecting virus particles are limited by their inability to sensitively detect small numbers of virus particles, particularly those that cannot be cultivated, and often provide false positives or negatives due to contamination and the need for complex calibration, while also failing to differentiate between virus subspecies and detect intact particles.
Innovation Solution
A method involving immobilizing capture molecules on a substrate, bringing virus particles into contact with these molecules, and then using probes that emit a specific signal for spatially resolved detection via fluorescence microscopy, allowing for quantitative and qualitative analysis of virus particles, including differentiation between intact and empty virus envelopes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If electron microscopy is used to detect virus particles, then direct detection of virus particles is enabled, but the method requires expensive equipment, large numbers of virus particles, and complex sample preparation
Solution Approach 1:
The patent introduces antibody-based capture molecules and detection probes as intermediaries between the virus particles and the detection system. These biomolecular mediators enable specific binding and signal generation without requiring complex electron microscopy equipment, thus resolving the contradiction between direct detection capability and equipment complexity
Solution Approach 2:
The patent replaces the mechanical/optical system of electron microscopy with a biochemical assay system based on antibody-antigen binding and fluorescence detection. This substitution eliminates the need for expensive electron microscopy equipment while maintaining the ability to directly detect virus particles
2Measurement precision
If PCR-based methods are used to detect viruses, then viral DNA/RNA can be amplified and detected, but the methods are highly susceptible to contamination causing false positives and require complex calibration
Solution Approach 1:
The patent extracts the detection target from genomic material to intact virus particles. By detecting whole particles rather than amplifying nucleic acids, the method eliminates the contamination susceptibility inherent in PCR while maintaining high detection sensitivity through specific antibody binding
Solution Approach 2:
The patent uses antibody binding as a specific recognition mechanism that does not involve amplification. The capture molecules and probes specifically bind to viral epitopes without requiring DNA/RNA amplification, thereby eliminating false positives from contamination while maintaining sensitivity
3Quantity of substance
If genome-based techniques are used to quantify viruses, then the amount of viral RNA/DNA can be measured, but the amount of viral particles cannot be determined
Solution Approach 1:
The patent creates a direct correspondence between the detection signal and individual virus particles through antibody binding. Each bound probe-capture molecule complex represents one virus particle, enabling absolute quantification of particle numbers rather than just nucleic acid amount
4Measurement precision
If ELISA tests are used to detect viruses, then viral fragments can be detected, but intact virus particles cannot be distinguished
Solution Approach 1:
The patent uses multiple antibodies with different specificities (capture molecules and detection probes) that bind to different epitopes on the virus particle. This localized binding at specific sites preserves and detects information about the intact particle structure, unlike methods that only detect fragmented viral components
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 ultra-sensitive detection of virus particles in any sample, including those that cannot be cultivated, with direct and absolute quantification and characterization, reducing false results and improving diagnostic accuracy by distinguishing between virus subspecies and detecting intact particles.
Implementation Method 1
immobilizing the virus particles on the substrate by binding to capture molecules, wherein the capture molecules are covalently bound to the substrate
Implementation Method 2
the probes are capable of emitting a specific signal, and steps b) and d) can be performed simultaneously or d) before b), wherein a spatially resolved determination of the probe signal is carried out.where detection is carried out using spatially resolved fluorescence microscopy
Data Source
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AI summary
The invention relates to a method for the quantitative and/or qualitative determination of virus particles containing at least one binding site for a capture molecule and at least one binding site for a probe. The invention further relates to a kit for carrying out said method and to various applications.