Nuclease-Cleavage Diagnostic Device for Point-Mutation Detection
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Solution Overview
Problem
Existing diagnostic technologies, such as PCR and CRISPR-based methods, face challenges in detecting small genetic variations and require laboratory equipment, lacking specificity and sufficient signal amplification.
Innovation Solution
A diagnostic device with reaction chambers, a membrane, and a detector for detecting a target nucleic acid molecule, utilizing a nuclease to cleave nucleic acid molecules bonded to markers, allowing for signal amplification and separation of cleaved portions through a porous membrane or magnetic beads.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If PCR amplification is used on a LOC, then amplification of nucleic acid sequences is achieved, but specificity to detect small changes such as point mutations is insufficient
Solution Approach 1:
The detection system is segmented into multiple functional chambers (reaction chamber, separation chamber, detection chamber) that perform distinct operations. The nuclease cleavage step is separated from amplification, allowing each process to be optimized independently for both quantity generation and specificity.
Solution Approach 2:
A bulky molecule acts as an intermediary carrier that temporarily binds the target nucleic acid during cleavage, then is separated away to allow detection of the cleaved fragments. This intermediary enables the cleavage reaction to occur with high specificity while allowing subsequent detection of the cleaved products.
2Quantity of substance
If the number of amplified sequences is increased to produce a detectable signal, then signal amplification is achieved, but the methods require laboratory equipment and facilities
Solution Approach 1:
Multiple functions (amplification, cleavage, separation, detection) are merged into a single integrated LOC device with microfluidic channels connecting compact chambers. This consolidation eliminates the need for separate laboratory equipment while maintaining all necessary functions for signal amplification and detection.
Solution Approach 2:
A porous membrane is used in the separation chamber to physically separate the bulky molecule from the cleaved nucleic acid fragments based on size differences. This simple porous barrier enables separation without requiring complex centrifugation or filtration equipment.
3Measurement precision
If a membrane with appropriate porosity is used to filter the first portion from the solution containing the second portion, then separation of cleaved portions is achieved, but device complexity increases
Solution Approach 1:
The porous membrane performs separation automatically based on the inherent size difference between the bulky molecule and the cleaved nucleic acid fragments. No external control or complex mechanism is needed - the size-based filtration occurs passively as fluid flows through the membrane, making the system self-regulating.
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 rapid, accurate, and cost-effective on-site detection of nucleic acid molecules without laboratory equipment, allowing for simultaneous detection of multiple pathogens or genetic variants from a single sample.
Implementation Method 1
a membrane in liquid connection to the at least one reaction chamber, having porosity sized to filter the first portion from the solution containing the second portion
Implementation Method 2
at least one reaction chamber, comprising a nuclease for cleaving a nucleic acid molecule
Data Source
AI summary
Diagnostic devices comprising: an inlet for receiving a solution comprising a biological sample comprising a target nucleic acid molecule; at least one reaction chamber, comprising a nuclease for cleaving a nucleic acid molecule, into a first portion bonded to a marker and a second portion bonded to a bulky molecule; a membrane in liquid connection to the at least one reaction chamber, having porosity sized to filter the first portion from the solution containing the second portion; and an informative chamber configured to receive the filtered first portion, having at least one of, a transparent region showing a visible marker; and a detector for detecting the marker are provided. Methods of using the devices to detect a target nucleic acid molecule in a sample are also provided.