Spatially Separated Reagent Drying in PCR Assay Containers
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Solution Overview
Problem
Existing methods for stabilizing biological and chemical detection reagents in dry form face instability issues when mixed together, particularly with nucleic acid polymerases, leading to adverse reactions and reduced assay accuracy, and require complex manufacturing processes or expensive equipment for liquid handling.
Innovation Solution
A reagent container with spatially separated areas for drying reagents, where nucleic acid polymerases and their substrates are dispensed distinctly to prevent adverse interactions, allowing for air-drying instead of lyophilization and reducing the need for insulating layers.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If detection reagents are dried as a mixture to simplify the manufacturing process, then manufacturing complexity is reduced, but reagent stability deteriorates due to adverse reactions between components
Solution Approach 1:
The reagent mixture is segmented into spatially separated zones within the reaction well. Each zone contains specific reagents that are physically isolated from others during drying and storage. The segmented zones are defined by hydrophobic barriers that prevent mixing, allowing stable drying of individual reagent components in their designated areas.
Solution Approach 2:
Hydrophobic barrier structures are introduced as intermediary elements between different reagent zones. These barriers act as mediators that prevent direct contact and adverse reactions between incompatible reagents while allowing each reagent to be dried and stored in its own protected zone within the same reaction well.
2Stability of the object's composition
If an insulating layer is added between reagents to prevent adverse reactions, then reagent stability is improved, but device complexity increases due to additional manufacturing steps
Solution Approach 1:
The insulating barrier function is merged with the reaction well structure itself. The hydrophobic barriers are integrated directly into the well architecture, eliminating the need for separate insulating layers. This integration maintains reagent stability while reducing manufacturing complexity by combining multiple functions into a single structure.
Solution Approach 2:
The reaction well structure is designed to serve multiple functions: it contains the reagents, provides hydrophobic barriers for zone separation, and facilitates drying and storage. This multi-functional design eliminates the need for additional insulating components, reducing device complexity while maintaining stability.
3Stability of the object's composition
If lyophilization is used to remove excess water from reagents, then reagent stability is improved, but manufacturing complexity increases due to technically demanding processes
Solution Approach 1:
The invention employs a simple air-drying approach instead of complex lyophilization equipment. The reaction wells are designed to be disposable or single-use, allowing simple evaporation of excess water at ambient conditions. This eliminates the need for expensive and complex lyophilization apparatus while maintaining adequate reagent stability for the intended application lifecycle.
4Stability of the object's composition
If small volume dispensing is used to avoid dissolving the insulating layer, then reagent stability is maintained, but measurement precision deteriorates due to variation in dispensing
Solution Approach 1:
The reaction well is designed with zones of different hydrophobicity. The barrier zones have high hydrophobicity to repel aqueous reagent solutions, while the reagent dispensing zones have hydrophilic properties to facilitate complete evaporation. This local differentiation of properties allows larger dispensing volumes without dissolving barriers, improving dispensing accuracy while maintaining stability.
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 method stabilizes reagents during drying and storage, enhancing assay accuracy and simplifying the manufacturing process by preventing unwanted reactions and allowing for more reliable dry reagent use in assays like PCR and immuno-PCR.
Implementation Method 1
said first and second zones being separated from each other by a hydrophobic barrier
Implementation Method 2
wherein excess water is removed from the reagents
Data Source
AI summary
A reagent container having an inner surface upon which at least two reagents are dried, with the first reagent dried on a first area separate from a second area where the second reagent is dried. The first and second reagents are a nucleic acid polymerase and its substrate. A method is disclosed which includes dispensing at least the first and second reagents onto separate areas of the inner surface of the reagent container, and removing excess water from the reagents. The reagent container can be used in a polymerase chain reaction (PCR) assay, an assay that utilizes a reverse transcriptase, a reverse transcriptase polymerase chain reaction, an immuno-PCR assay, a nucleic acid sequence based assay, a proximity ligation assay, a ligase chain reaction assay, a rolling circle amplification assay, and a strand displacement amplification assay.