Microfluidic Device for Naked-Eye Gene Detection via Isothermal RCA
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Solution Overview
Problem
Current gene detection methods for pathogens are time-consuming, require expensive equipment, and struggle with accuracy and the ability to detect multiple pathogens simultaneously, often needing external power and specialized devices like thermal cyclers.
Innovation Solution
A microfluidic device with a dumbbell-shaped template and immobilized primer, capable of rolling circle amplification at room temperature, allowing for naked-eye detection of target genes without external power or specialized devices, using a microfluidic device with a board, inlet, first and second channels, and a coating for primer immobilization.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If PCR-based detection methods are used, then detection accuracy is improved, but detection time and cost increase significantly
Solution Approach 1:
The invention changes the fundamental detection parameter from temperature-cycling-based PCR to isothermal rolling circle amplification (RCA). By maintaining a constant temperature (37°C) instead of cycling through multiple temperatures, the method achieves both high detection accuracy through specific template-primer binding and significantly reduced detection time without requiring thermal cyclers
Solution Approach 2:
The invention extracts and removes the thermal cycler requirement from the detection system. By using isothermal amplification instead of PCR, the complex temperature cycling equipment is eliminated, retaining only the essential detection functionality while reducing time and cost
2Reliability
If thermal cycler equipment is used, then PCR reaction conditions are met, but device complexity and cost increase
Solution Approach 1:
The invention extracts and eliminates the thermal cycler from the system. By using isothermal rolling circle amplification at 37°C, the complex temperature cycling equipment is removed entirely, simplifying the device while maintaining reliable reaction conditions through constant temperature incubation
Solution Approach 2:
The invention replaces expensive, complex thermal cycler equipment with simple, inexpensive isothermal reaction conditions. The microfluidic device can be a simple chip or even a disposable format, eliminating the need for costly specialized equipment while achieving reliable detection
3Measurement precision
If conventional PCR methods are used, then target gene amplification is achieved, but the ability to detect multiple pathogens simultaneously is limited
Solution Approach 1:
The invention segments the detection system into multiple independent channels or reaction zones within the microfluidic device. Each channel can contain different templates specific to different pathogens, allowing simultaneous amplification and detection of multiple targets through parallel isothermal RCA reactions
Solution Approach 2:
The invention creates a universal microfluidic platform that can detect multiple pathogens simultaneously. By using the same isothermal RCA mechanism with different pathogen-specific templates, the system achieves multiplex detection capability while maintaining the simplicity of a single device design
4Measurement precision
If external power supply and specialized devices are used, then detection sensitivity is improved, but ease of operation and portability are reduced
Solution Approach 1:
The invention extracts and removes the requirement for external power supplies and complex specialized devices. By using isothermal amplification at 37°C (body temperature), the system can operate without electricity, using only simple incubation conditions that can be achieved with passive heating or even body heat, dramatically improving portability and ease of operation
Solution Approach 2:
The invention enables the detection system to be self-sufficient without external power. The isothermal reaction can proceed using ambient or body temperature, and the microfluidic device can be designed as a standalone unit requiring no external equipment, making it suitable for field deployment and point-of-care testing
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, cost-effective, and accurate detection of single or multiple target genes, including pathogens, at room temperature without external power or specialized equipment, facilitating quick diagnosis and broad applicability for infectious diseases and bioterrorism.
Implementation Method 1
a template complementarily binding to the primer, wherein the template includes a binding region complementary to a target gene
Implementation Method 2
a primer immobilized on the coating of the second channel
Implementation Method 3
a combination of surface-associated molecular padlock DNA probes (MPPs) and rolling circle amplification (RCA)
Data Source
Figure 1~2E
Figure 3A~3F
Figure 4~5
AI summary
The present invention provides a target gene capable of being differentiated by the naked eye by amplifying the target gene to selectively block the fluid path and, specifically, a microfluidic device for detecting pathogen genes, and a detection method using the same. Therefore, the present invention can conveniently detect a single target gene, such as a single pathogen, or at the same time, several target genes, such as several pathogens, without complicated mechanical devices.