PCR Primer Microdot Array for Rapid Gene Detection
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Solution Overview
Problem
Current Lab-on-Chip devices for gene amplification are costly, time-consuming, and require multiple cycles of amplification, hybridization, and washing steps, necessitating special preparation and sterilization to ensure proper PCR amplification.
Innovation Solution
A substrate with microdots containing primers for gene amplification, stabilized for extended periods, allows for concurrent gene amplification and fluorescence sensing during RT-qPCR, enabling rapid detection of target genes using fluorophores and excitation energy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If multiple cycles of amplification, hybridization, and washing steps are performed in current Lab-on-Chip devices, then gene amplification accuracy is improved, but reaction time and operational complexity increase
Solution Approach 1:
Primers are pre-stabilized within microdots on the substrate before the PCR process begins. This preliminary stabilization eliminates the need for multiple washing steps to secure primers during amplification, allowing direct amplification and detection without time-consuming intermediate steps while maintaining amplification accuracy
Solution Approach 2:
The invention combines the amplification process with concurrent fluorescence detection in a single integrated step. The stabilized primers in microdots enable real-time fluorescence sensing during amplification, merging what were previously separate processes (amplification cycles followed by hybridization and washing) into one simultaneous operation, thereby reducing total reaction time
2Reliability
If special preparation and sterilization of reaction chambers are performed, then PCR amplification reliability is improved, but device complexity and cost increase
Solution Approach 1:
The invention uses a disposable substrate with pre-stabilized primers in microdots that can be discarded after a single use. This eliminates the need for complex sterilization and repeated preparation of reaction chambers, as each new substrate is already prepared and sterile-ready. The low-cost disposable nature reduces both device complexity and operational costs while maintaining reliable amplification
Solution Approach 2:
All necessary preparation including primer stabilization and sterilization is performed in advance during substrate manufacturing. The substrate arrives ready-to-use without requiring on-site preparation or sterilization steps, simplifying the device operation and reducing complexity while ensuring reliable amplification conditions
3Measurement precision
If multiple handling and washing steps are performed, then detection accuracy is improved, but ease of operation decreases
Solution Approach 1:
Primers are pre-stabilized within the microdots on the substrate before use. This preliminary stabilization ensures that primers remain firmly attached during the entire PCR process without requiring intermediate washing steps to secure them, thereby maintaining detection accuracy while significantly simplifying the operational steps required
Solution Approach 2:
The stabilized primers in microdots perform their function automatically during the amplification process without requiring external intervention for washing or repositioning. The system essentially serves itself by maintaining primer integrity through stabilization, eliminating the need for operator-performed washing steps while preserving detection precision
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 solution provides a cost-effective, rapid, and reliable method for gene detection by stabilizing primers in microdots, allowing for immediate fluorescence-based results without extensive handling, reducing reaction time and costs.
Implementation Method 1
Each microdot contains one or more primers for gene amplification... the primer will hybridize to a particular target gene to be tested
Implementation Method 2
The fluid also contains fluorophores which will be fixed into the gene as intercalating fluorophores... the microdots are subjected to an excitation energy, such as by applying a laser light, UV light, or other acceptable excitation energy that will cause the fluorophores to output light
Data Source
AI summary
A substrate has a plurality of microdots positioned thereon. Each microdot contains one or more primers for gene amplification for a particular target gene. The microdots are placed on the substrate and the substrate is positioned in a housing. The housing has a sample fluid to be tested introduced therein covering the microdot array. While the sample fluid is overlying the substrate, the amplification of the target gene is carried out if it is present within the sample. If the target gene that matches the primers is not present, then amplification will not take place. The fluid also contains fluorophores which will be fixed into the gene as it increases in size as it clearly detects if gene amplification has occurred by detecting the amount of light detected for a particular microdot. In a preferred embodiment, the sample fluid is placed on top of a sealing layer that is less dense then water, such as wax or mineral oil. During a heating of the sample fluid and sealing layer, the sample fluid will sink to the bottom of the sealing layer so that it is fully encased and protected.