Multiplex Target Particle Analysis via Segmented Probe Amplification
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Solution Overview
Problem
Current methods lack the capability to comprehensively analyze target particles in specimens simultaneously, particularly extracellular vesicles, which are crucial for disease detection and research due to their involvement in cancer and other diseases, but their configuration and function are not well understood.
Innovation Solution
A method involving the use of probes with specific binding portions and nucleic acid reporter portions, including common and specific amplification sequences, to capture, amplify, and analyze target particles by detecting surface markers, allowing for the simultaneous analysis of multiple target particles in a specimen.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If conventional analysis methods are used for target particles, then the analysis process is simple, but the capability to comprehensively analyze multiple target particles simultaneously is insufficient
Solution Approach 1:
The analysis method is divided into distinct sequential steps: capture step (capturing target particles using capture probes), amplification step (amplifying captured particles using amplification probes with polymerase), and detection step (detecting amplified particles). This segmentation allows each step to be optimized independently while achieving comprehensive simultaneous analysis of multiple target particles through parallel processing in multiple reaction vessels.
2Productivity
If multiple target particles are analyzed simultaneously, then the productivity and comprehensiveness of analysis is improved, but the complexity of the analysis method increases
Solution Approach 1:
The system employs universal probe structures with standardized regions (capture region, spacer region, amplification region) that can bind to different target particles through specific sequences while maintaining consistent amplification and detection protocols. This multi-functionality allows the same basic methodology to analyze multiple different target particles simultaneously across multiple reaction vessels, increasing productivity without proportionally increasing method complexity.
Solution Approach 2:
Amplification probes serve as intermediaries that bridge the capture step and detection step. These probes contain both polymerase binding sites for amplification and detection sequences, enabling the transformation of captured target particles into detectable signals through a standardized amplification process that works across multiple different target particle types.
3Loss of information
If comprehensive analysis of target particles is performed, then information on disease detection and research is improved, but the complexity of probe design and analysis procedure increases
Solution Approach 1:
The probe design incorporates local quality differentiation through distinct functional regions: capture regions with specific sequences for different target particles, spacer regions for structural stability, and amplification regions with standardized polymerase binding sites. This allows each probe to be specifically tailored for its target while maintaining a universal amplification and detection framework, reducing overall system complexity.
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 the comprehensive analysis of target particles, providing information on their types, quantities, and surface markers, which can be used for disease diagnosis and research, facilitating early disease detection and prognosis.
Implementation Method 1
a binding portion that specifically binds to any one of first to m-th surface markers of the target particle
Implementation Method 2
individually amplifying the reporter portion for each of the first1 to firstn containers using the common primer set to obtain first to n-th amplification products
Data Source
AI summary
According to one embodiment, a method includes dispensing the specimen into first1 to firstn containers configured to capture the target particle, removing a contaminant other than the target particle to be captured from the specimen, adding first to m-th probes to the first1 to firstn containers, removing excessive first to m-th probes that have not bound to the target particle, individually amplifying the reporter portion for each of the first1 to firstn containers using the common primer set to obtain first to n-th amplification products, removing an excessive common primer set from the first to n-th amplification products, dispensing the first to n-th amplification products into second1 to secondm containers respectively, amplifying the amplification products in the second1 to secondm containers using the first to m-th specific primer sets, and analyzing presence or absence or types of the target particles captured in the first1 to firstn containers.