Microwell Array for Rapid Pathogen Detection
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Solution Overview
Problem
Current diagnostic techniques for detecting low concentration pathogens in fluid samples are time-consuming, costly, and lack sensitivity, often requiring multiple tests and relying on outdated methods like cell cultures and microscopy, which can lead to misdiagnoses and antibiotic overuse, especially in conditions like sepsis.
Innovation Solution
The development of a system and method using an array of wells for nucleic acid detection, including amplification techniques, that allows for rapid and sensitive detection of pathogens in very low concentrations within fluid samples, including bodily fluids, food, and environmental samples, by drying the sample and applying reagents to each well for bi-phasic reactions, enabling detection of multiple targets in a single assay.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional diagnostic techniques (cell cultures, microscopy, immunoassays) are used, then detection can be performed with simple equipment, but sensitivity is insufficient and detection time is too long (days to a week)
Solution Approach 1:
The sample is partitioned into an array of wells, creating numerous isolated reaction chambers. This segmentation allows parallel processing of multiple samples and enables detection of low-concentration pathogens by concentrating target nucleic acids in individual wells, thereby improving sensitivity while reducing overall detection time through parallelization
Solution Approach 2:
The method performs nucleic acid amplification directly in the array wells without requiring prior purification steps. By preparing the reaction mixtures in advance and loading them into the array, the system eliminates time-consuming intermediate purification steps while maintaining detection sensitivity
2Measurement precision
If nucleic acid amplification techniques are used, then detection sensitivity improves, but equipment cost and operational complexity increase
Solution Approach 1:
The array platform serves multiple functions: it holds samples, performs amplification reactions, and enables detection. The same physical structure supports various nucleic acid amplification methods (PCR, LAMP, RPA), making the system versatile and reducing the need for specialized equipment for each technique
Solution Approach 2:
The method uses nucleic acid copying through amplification techniques to generate multiple copies of target sequences from minimal initial material. This allows detection of low-concentration pathogens without requiring expensive high-sensitivity instrumentation, as the amplification process itself creates detectable signal levels
3Measurement precision
If sample purification steps are performed before analysis, then detection accuracy improves, but processing time and cost increase
Solution Approach 1:
The system performs amplification reactions directly in the array wells using pre-prepared reaction mixtures, eliminating the need for intermediate purification steps. The primers and enzymes are loaded in advance, allowing direct amplification from crude samples and significantly reducing processing time while maintaining detection accuracy
Solution Approach 2:
The method combines sample loading, amplification, and detection into a single integrated process within the array. By merging these steps and eliminating separate purification stages, the system achieves rapid processing without sacrificing detection accuracy, as the amplification reaction itself tolerates crude sample conditions
4Measurement precision
If single-pathogen testing is performed, then test specificity is high, but multiple separate tests are required increasing overall time and cost
Solution Approach 1:
The array is divided into multiple wells, each capable of testing for different pathogens using pathogen-specific primers. This segmentation allows simultaneous testing for multiple pathogens in parallel, maintaining high specificity for each target while reducing total testing time compared to sequential single-pathogen tests
Solution Approach 2:
The same array platform and amplification methodology can detect multiple different pathogens by simply changing the primer sets. This multi-functionality allows a single test system to replace multiple separate tests, reducing overall time and cost while maintaining the specificity needed for accurate pathogen identification
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 enables rapid detection of multiple pathogens in under 45 minutes, reducing diagnosis time and costs, improving accuracy, and allowing for quicker treatment by detecting pathogens at very low concentrations without the need for extensive sample preparation or purification.
Implementation Method 1
nucleic acid detection, including amplification techniques
Implementation Method 2
detecting a target nucleic acid if present in a dried sample island by a bi-phasic reaction
Data Source
AI summary
Described herein are systems and methods which utilize an array of wells to isolate pathogens and nucleic acid detection techniques to accurately and rapidly detect pathogens in fluid samples, even in very low concentrations, including from solid or semi-solid samples that have been fluidized. The provided systems and methods dry the fluid sample to deposit a fraction of the total volume in a number of wells and perform nucleic acid detection on individual wells to detect even individual pathogens and provide a quantitative analysis of the amount of pathogen within the sample. Also provided are methods and systems for precise delivery of dried materials, including biomolecules that are enzymes of use in the process, to microwells.