Self-contained pouch for multiplex nucleic acid analysis
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Solution Overview
Problem
Traditional microbiology techniques for diagnosing infectious diseases are time-consuming, and methods like PCR and immuno-PCR face challenges with contamination and robustness, especially when dealing with multiple pathogens and low nucleic acid concentrations, requiring complex handling and increasing the risk of contamination.
Innovation Solution
A self-contained, flexible pouch system for nucleic acid analysis that allows for nested PCR and immuno-PCR, minimizing contamination through a closed system with multiple reaction zones and channels, enabling robust amplification and detection of multiple biological substances in a single assay.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If traditional microbiology techniques are used for diagnosing infectious diseases, then the diagnosis can be performed with simple equipment, but the diagnosis takes days or weeks, delaying treatment
Solution Approach 1:
The patent replaces traditional mechanical microbiology techniques (culturing, staining) with molecular biology techniques (PCR amplification). This substitution enables rapid detection of pathogen nucleic acids within hours rather than days, dramatically reducing diagnosis time while maintaining high accuracy through specific primer binding and amplification.
Solution Approach 2:
The patent changes the detection parameter from detecting visible microbial structures (requiring culture growth) to detecting molecular nucleic acid sequences. This parameter change allows direct detection of pathogens in clinical samples without culture requirements, enabling rapid diagnosis within hours while maintaining specificity through sequence-based identification.
2Adaptability or versatility
If large panels of PCR assays are run to diagnose multiple possible causative organisms, then the diagnostic coverage is improved, but the complexity and cost increase significantly
Solution Approach 1:
The patent merges multiple individual PCR assays into a single multiplex PCR reaction by incorporating multiple primer pairs targeting different pathogens into one reaction tube. This combining approach maintains the ability to detect multiple causative organisms simultaneously while dramatically reducing operational complexity from running many separate assays to executing one integrated reaction.
Solution Approach 2:
The patent creates a universal diagnostic platform where a single PCR reaction system can detect multiple different pathogens through the inclusion of multiple primer sets. This multi-functionality allows one assay system to serve multiple diagnostic purposes, improving adaptability without proportionally increasing complexity.
3Productivity
If multiplex PCR is used to assay for multiple targets concurrently, then the efficiency is improved, but the robustness of high level multiplex reactions decreases and analysis of multiple products becomes difficult
Solution Approach 1:
The patent segments the multiplex PCR analysis into distinct functional zones within a single reaction system. Different primer pairs and target sequences are organized in spatially separated regions or with distinct fluorescent labels, allowing simultaneous amplification of multiple targets while maintaining analytical resolution. This segmentation prevents product interference and maintains reaction robustness despite high multiplexing.
4Measurement precision
If nested secondary PCRs are performed to increase robustness, then the detection sensitivity is improved, but the handling complexity and contamination risk increase
Solution Approach 1:
The patent merges the nested PCR steps into a single simultaneous multiplex reaction rather than performing sequential nested reactions. Multiple primer pairs including nested primers are included in one reaction tube, achieving the enhanced detection sensitivity of nested PCR while eliminating the handling complexity and contamination risks associated with multiple sequential steps.
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 system provides a rapid, sensitive, and robust method for diagnosing infectious diseases by minimizing contamination and ensuring robust amplification, enabling the detection of multiple pathogens in a single assay with reduced handling and increased efficiency.
Implementation Method 1
a first-stage reaction blister configured for first-stage amplification of the sample
Implementation Method 2
each second-stage reaction chamber comprising a pair of primers configured for further amplification of the sample
Implementation Method 3
the blisters comprise a flexible material, such that pressure provided on an individual blister collapses the blister, forcing the contents from the blister
Implementation Method 4
the first stage reaction zone is an antigen-binding zone for immuno-PCR, in which antigens present in the sample are recognized and associated with a particular nucleic acid segment
Data Source
AI summary
Devices, containers, and methods are provided for performing biological analysis in a closed environment. Illustrative biological analyses include high density nucleic acid amplification and detection and immuno-PCR.