Multi-Wavelength CRISPR Pathogen Detection in Single Chamber
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Solution Overview
Problem
Current methods for detecting pathogens in biological samples are often time-consuming, require multiple channels for each target pathogen, and may not be suitable for point-of-care testing in low-complexity medical environments.
Innovation Solution
The use of multi-wavelength sensing with spectrally unique loop-mediated isothermal amplification (LAMP) primers and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) enzymes tagged with fluorescent probes allows for simultaneous detection of multiple target pathogens in a single fluorescence reading chamber.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple independent channels are used for detecting each target pathogen, then detection accuracy is improved, but device complexity increases
Solution Approach 1:
The patent combines multiple pathogen detection channels into a single fluorescence reading chamber. Multiple CRISPR-Cas systems with different fluorescent probes (e.g., FITC for SARS-CoV-2, TRITC for Influenza A) are integrated into one chamber, allowing simultaneous detection of multiple pathogens without requiring separate independent channels for each pathogen.
Solution Approach 2:
The single fluorescence reading chamber is designed to perform multiple detection functions simultaneously. The system uses a universal reading chamber that can detect multiple different pathogens by employing spectrally distinct fluorescent probes, making the chamber multi-functional rather than dedicated to a single pathogen detection task.
2Reliability
If multiple independent channels are used for each target pathogen, then detection reliability is improved, but ease of operation deteriorates
Solution Approach 1:
By merging multiple detection channels into one reading chamber, the system reduces the operational steps required for testing. Users no longer need to operate multiple separate channels or devices, simplifying the testing procedure and making it more suitable for point-of-care environments where ease of operation is critical.
Solution Approach 2:
The universal reading chamber design allows a single device to handle multiple pathogen detections, improving ease of operation by eliminating the need for multiple specialized devices or complex multi-step procedures across different channels, while maintaining reliable detection through spectrally resolved fluorescence measurement.
3Device complexity
If a single fluorescence reading chamber is used, then device complexity is reduced, but measurement precision deteriorates
Solution Approach 1:
The system employs spectrally distinct fluorescent probes that emit at different wavelengths (e.g., FITC emits green fluorescence, TRITC emits red fluorescence). By detecting these different emission wavelengths within the single reading chamber, the system maintains the ability to precisely distinguish and quantify multiple pathogens simultaneously, preserving measurement precision despite using a single chamber.
Solution Approach 2:
The system transitions from spatial separation (multiple independent channels) to spectral separation (different wavelengths) as the distinguishing dimension. By measuring fluorescence emissions at multiple wavelengths within a single chamber, the system achieves multi-pathogen detection precision without requiring multiple physical channels, effectively adding a spectral dimension to the detection process.
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 reduces the number of independent channels needed for point-of-care testing, enables real-time detection, and is adaptable for use in various environments, including low-complexity medical settings.
Implementation Method 1
fluorescent probes allows for simultaneous detection of multiple target pathogens
Data Source
AI summary
Systems and methods for testing a biological sample for each of two or more target pathogens employ fluorescence detection from a single fluorescence reading chamber. A method of testing a biological sample for each of two or more target pathogens includes forming a detection cycle liquid by combining detection cycle compounds with a biological sample liquid formed from a biological sample. The detection cycle compounds include respective CRISPR enzymes for each of the two or more target pathogens. The respective CRISPR enzymes are tagged with respective spectrally unique fluorescent probes. The detection cycle liquid is exposed to a multi-wavelength fluorescence excitation light. A multi-wavelength fluorescence light emission detection signal indicative of intensity of fluorescence light emitted by the detection cycle liquid over multiple wavelengths is processed to generate detection data indicative of presence or absence of each of the two or more target pathogens in the biological sample.