Nucleic Acid Detection Dye Color Shift for Visual Analysis
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting nucleic acid amplification products are complex, require specialized equipment, and suffer from reduced sensitivity due to background signals and high costs associated with fluorescence-labeled reagents.
Innovation Solution
A method and kit utilizing dyes whose color tone changes upon binding to nucleic acids, allowing for visual detection without the need for specialized devices, by using substances that react with bound or unbound dyes to enhance contrast and simplify the detection process.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If fluorescent intercalators are added to reaction liquid before PCR and fluorescence intensity is measured with a fluorospectrophotometer, then the amount of amplified DNA can be determined without electrophoresis, but background signal from single-stranded nucleic acid binding enhances the signal and reduces detection sensitivity
Solution Approach 1:
The invention extracts and removes single-stranded nucleic acids from the reaction mixture before detection. By eliminating the source of background signal (single-stranded nucleic acids that bind fluorescent intercalators), the method preserves detection speed while improving detection sensitivity through reduced background interference
Solution Approach 2:
The invention introduces an intermediary step (separation or selective removal mechanism) between amplification and detection. This intermediary process selectively removes single-stranded nucleic acids while preserving double-stranded amplification products, allowing accurate fluorescence-based detection without background interference
2Measurement precision
If fluorescence-labeled primers are used and fluorescence polarization detection is performed, then nucleic acid amplification products can be detected, but complicated separation operations are required to remove unincorporated primers, reducing yield and detection sensitivity
Solution Approach 1:
The invention extracts unincorporated (free) primers from the reaction mixture before detection. By removing these interfering substances through selective precipitation or other separation methods, the system achieves accurate detection without requiring complex purification equipment or procedures
Solution Approach 2:
The invention employs simple, disposable separation methods (such as ethanol precipitation or magnetic bead-based cleanup) instead of expensive, complex purification systems. These straightforward approaches effectively remove free primers while preserving amplification products, maintaining detection sensitivity without adding operational complexity
3Measurement precision
If angle of rotation or circular dichroism of polarized light is measured to detect nucleic acid amplification products, then detection can be performed, but special devices are required making the method complex
Solution Approach 1:
The invention replaces complex optical measurement systems (polarized light rotation or circular dichroism devices) with a simpler fluorescence-based detection system. By using fluorescent intercalators and standard fluorospectrophotometers, the method achieves equivalent or superior detection capability with widely available, inexpensive equipment
Solution Approach 2:
The invention changes the detection parameter from optical rotation or circular dichroism to fluorescence intensity measurement. This parameter change enables the use of simpler, more common laboratory equipment while maintaining or improving detection sensitivity and ease of operation
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 simple and precise visual detection of nucleic acid amplification products using visible light, reducing costs and eliminating the need for expensive equipment, while improving detection sensitivity and speed.
Implementation Method 1
a dye which binds to nucleic acid... the color tone of which changes from blue to green
Implementation Method 2
contacting a substance which reacts with the dye, specifically an oxidizing agent, a reducing agent, an acid, a base or a pH buffering agent
Data Source
AI summary
A method and a device or kit for detecting a nucleic acid, which enable simple and precise visual detection of a nucleic acid amplified by an nucleic acid amplification method, without necessity of special devices are provided. The method for detecting a nucleic acid in a sample comprises: contacting a sample with a dye to react with each other; and observing a substance produced by the reaction with visible light, and evaluating the presence or absence of a nucleic acid by eye. The device or kit for detecting a nucleic acid in a sample comprises: a carrier that holds a dye which can bind to a nucleic acid; a path for passing a sample through the carrier; and an evaluation part for observing a substance produced by the reaction between the sample and the dye with visible light, and evaluating the presence or absence of a nucleic acid by eye.