Virion Capture and Detection via Aptamer-FISH Integration
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Solution Overview
Problem
Current methods for virus detection, such as ELISA and PCR, are either rapid but not sensitive or sensitive but not rapid, and they do not directly measure the concentration of infectious viral particles.
Innovation Solution
A method involving the capture of virions on a solid substrate using aptamers, followed by fixation, washing, and detection using fluorescence in situ hybridization (FISH), allowing for the direct detection of infectious virions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If ELISA-based detection of viral surface antigen is used, then detection speed is rapid, but sensitivity is insufficient
Solution Approach 1:
The patent combines antigen capture (ELISA principle) with genome detection (FISH principle) into a single integrated assay. Virions are captured on solid substrate via antigen-binder interaction, then fixed and subjected to FISH for genome detection, merging the rapid capture advantage with the sensitive detection advantage in one workflow
Solution Approach 2:
The solid substrate acts as an intermediary that first captures whole virions through antigen-binder interaction, then allows FISH probes to access and detect the viral genome within the fixed virions. This intermediary approach enables both rapid capture and sensitive genome detection without requiring separate parallel assays
2Measurement precision
If PCR-based detection of viral genome is used, then detection sensitivity is high, but detection speed is slow
Solution Approach 1:
The virions are pre-captured and fixed on the solid substrate before genome detection. This preliminary capture and fixation step concentrates the viral particles and stabilizes them, enabling rapid FISH detection without requiring lengthy PCR amplification steps, thus achieving both sensitivity and speed
3Ease of operation
If surface antigen or viral genome detection is performed separately, then detection is indirect, but direct measurement of infectious virion concentration cannot be achieved
Solution Approach 1:
The assay merges antigen-based virion capture with genome-based detection in a single integrated workflow. By requiring both antigen presence (for capture) and genome presence (for FISH detection), the method directly measures intact infectious virions rather than indirect markers, achieving accurate infectious titer measurement
Solution Approach 2:
The method uses FISH probes that copy the principle of detecting intact viral particles by requiring both surface antigen (capture target) and internal genome (detection target) to be present simultaneously, creating a dual-verification system that accurately reflects infectious virion concentration
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 fast, sensitive, and direct assay for virus detection, enabling the determination of infectious titers without the need for cell-based assays, thus improving accuracy and consistency.
Implementation Method 1
capturing the virion on a solid substrate comprising, coating the solid substrate, immobilizing a binder to the solid substrate, and contacting the solid substrate with the virion, thereby capturing the virion on the solid substrate
Implementation Method 2
fixation is methanol fixation
Implementation Method 3
detecting the virion
Data Source
AI summary
Provided herein are, in various embodiments, methods and kits for assaying one or more virions. In certain embodiments, the methods and kits of the disclosure provide for the calculation of virion titer and/or virion infectivity. In still further embodiments, the disclosure provides for methods and kits for enhancing assaying of viruses such as SARS-CoV-2.