Nonplanar Solid Substrate for Integrated Cell Capture and Nucleic Acid Amplification
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Solution Overview
Problem
Conventional methods for amplifying nucleic acids from cells are time-consuming, complex, and not suitable for Lab-On-a-Chip (LOC) applications, requiring high-concentration reagents that can interfere with subsequent processes like PCR, and do not allow for simultaneous cell isolation and nucleic acid amplification in a single vessel.
Innovation Solution
A method involving a nonplanar solid substrate is used to attach cells to its surface in a liquid medium with a pH of 3.0-6.0, followed by washing and performing PCR in a single vessel to amplify nucleic acids, increasing surface area for cell binding and reducing solubility, thus enabling efficient nucleic acid isolation and amplification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional methods are used for nucleic acid isolation and amplification, then nucleic acids can be amplified, but the process is time-consuming and complex, requiring multiple separate steps for cell separation, nucleic acid isolation, and amplification
Solution Approach 1:
The patent combines cell separation, nucleic acid isolation, and amplification into a single integrated process performed in one vessel. The nonplanar solid substrate serves multiple functions: it captures cells through binding, enables nucleic acid extraction directly from the captured cells, and allows subsequent amplification reactions without transferring samples between different processing stations or vessels.
Solution Approach 2:
The nonplanar solid substrate acts as a universal platform that performs multiple functions: cell capture, nucleic acid binding, and reaction support. This multi-functional substrate eliminates the need for separate specialized equipment for each processing step, thereby simplifying the overall system while maintaining high productivity.
2Reliability
If high-concentration reagents are used for DNA binding to solid phase, then nucleic acids bind effectively, but the reagents interfere with subsequent PCR processes
Solution Approach 1:
The patent extracts or removes the harmful high-concentration reagents from the system after they have served their purpose of enabling nucleic acid binding. The washing step specifically removes chaotropic salts and other binding reagents from the solid phase, eliminating their inhibitory effect on subsequent PCR while preserving the bound nucleic acids.
Solution Approach 2:
The nonplanar solid substrate acts as an intermediary that facilitates nucleic acid binding without requiring harmful high-concentration reagents. By providing a large surface area with appropriate surface chemistry, the substrate enables effective binding under milder conditions that are compatible with subsequent PCR amplification.
3Quantity of substance
If conventional planar substrates are used for cell binding, then cells can be attached, but the surface area is limited, reducing binding capacity
Solution Approach 1:
The patent employs a nonplanar solid substrate with curved or three-dimensional surface features that provide significantly larger surface area compared to flat planar substrates. This increased surface area allows for greater cell binding capacity while maintaining a compact form factor suitable for integrated systems.
Solution Approach 2:
The invention transitions from two-dimensional planar surfaces to three-dimensional nonplanar structures. By utilizing vertical and lateral dimensions simultaneously, the substrate maximizes surface area within a confined space, thereby increasing cell binding capacity without proportionally increasing the overall device footprint.
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
This approach allows for efficient and rapid nucleic acid amplification, reducing the need for additional reagent removal steps and enabling high-sensitivity PCR, even in samples with inhibitors like SPS, while automating cell separation and amplification processes.
Implementation Method 1
contacting a sample comprising a cell with a nonplanar solid substrate in a liquid medium having a pH in a range of 3.0-6.0 to attach the cell to the nonplanar solid substrate
Data Source
AI summary
Provided is a method of amplifying nucleic acid from a comprising: contacting a cell-containing sample with a nonplanar solid substrate in a liquid medium having a pH range of 3.0-6.0 to attach the cell to the solid substrate; washing the nonplanar solid substrate to remove materials that are not attached thereto; and performing PCR using the nucleic acid from the cell attached to the nonplanar solid substrate as a template sample to amplify nucleic acid from the cell, wherein the contacting, washing and PCR processes are performed in a single vessel.