Molecular Beacon for Nucleic Acid Detection via Colorimetric Readout
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Solution Overview
Problem
Current diagnostic assays for nucleic acid detection are often bulky, expensive, and require electronic devices, limiting their use for point-of-need applications and home usage, especially since fluorescence-based methods require instrumentation for detection, while absorption-based assays lack sensitivity for nucleic acid detection.
Innovation Solution
A system comprising a translation module, amplification module, and detection module that uses nucleic acid peroxidases to provide a readable output, where the amplification module includes CRISPR-Cas13 or Cas12 modules to activate multiple peroxidase units per target nucleic acid strand, enabling sensitive detection without electronic devices.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescence-based methods are used for nucleic acid detection, then detection sensitivity is improved, but the requirement for electronic instrumentation increases
Solution Approach 1:
The patent replaces fluorescence-based optical detection with a colorimetric chemical detection system. Instead of using fluorophores that require excitation light sources and detectors, the invention employs peroxidase enzymes that catalyze color-changing reactions visible to the naked eye, substituting electronic/optical systems with simple chemical reactions
Solution Approach 2:
The patent utilizes colorimetric changes as the detection readout mechanism. Peroxidase enzymes catalyze reactions that produce colored products from colorless substrates, enabling visual detection without instrumentation. The system monitors color intensity changes to quantify nucleic acid presence and concentration
2Device complexity
If absorption-based assays are used for detection, then device simplicity is improved, but detection sensitivity deteriorates
Solution Approach 1:
The patent changes the detection parameter from direct absorption measurement to enzyme-catalyzed colorimetric signal amplification. By introducing peroxidase enzymes that amplify the signal through catalytic cycles, the system achieves high sensitivity with simple visual readout, transforming a low-sensitivity absorption assay into a high-sensitivity colorimetric assay
Solution Approach 2:
The patent introduces peroxidase enzymes as intermediary catalysts between the nucleic acid target and the detectable signal. These enzymes act as signal amplifiers that convert微量nucleic acid binding events into macroscopic color changes, bridging the gap between simple detection and high sensitivity requirements
3Productivity
If multiple peroxidase units are activated per target nucleic acid strand, then signal amplification is improved, but system complexity increases
Solution Approach 1:
The patent segments the detection system into distinct functional modules: a translation module that converts target nucleic acid into a recognizable format, an amplification module that activates multiple peroxidase units, and a detection module that reads the colorimetric signal. This segmentation allows complex signal amplification to be achieved through coordinated simple steps
Solution Approach 2:
The patent performs preliminary preparation of peroxidase units in an inactive state, embedded within nucleic acid structures that prevent their activity. Upon target detection, these pre-positioned enzymes are activated en masse, enabling rapid signal amplification without requiring complex real-time activation mechanisms
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
The system allows for sensitive and specific detection of nucleic acids with a naked-eye readable output, capable of detecting trace amounts of nucleic acids in a device-free format, enhancing accessibility for point-of-need applications and home usage.
Implementation Method 1
the reporter polynucleotide is configured to hybridize the target polynucleotide
Implementation Method 2
the molecular beacon comprises a peroxidase or a polynucleotide configured to encode the peroxidase
Implementation Method 3
the peroxidase or the activated peroxidase is configured to convert the substrate into a product
Implementation Method 4
Colorimetric Molecular Beacon
Data Source
AI summary
Systems comprising a translation module, an amplification module comprising one or more sensing modalities, and a detection module configured to accept the amplifier output and methods of using the same are provided. The systems or methods can be capable of detecting trace amounts of nucleic acid molecules with high sensitivity and specificity in a device-free system with naked-eye readable output. The systems can comprise three layers incorporated into a lateral flow assay and/or a paper strip assay, in such systems, the first layer can comprise the translation module, the second layer can comprise the amplification module, and the third layer can comprise the detection module.