LAMP Assay Cartridge Mixing for Inhibitor-Resistant Pathogen Testing
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Solution Overview
Problem
Conventional nucleic acid testing methods, such as PCR, are inefficient in geographical regions lacking complex laboratory equipment, and there is a need for improved pathogen detection using loop-mediated isothermal amplification (LAMP) that can perform at non-cyclical and relatively low temperatures.
Innovation Solution
An assay cartridge with a sample introduction area, mixing region, test well, and fluid path, utilizing magnetic fields, vibrations, or sonic generators to enhance mixing and amplification of samples, and incorporating agents to inhibit inhibitors like lactoferrin, lysozyme, nucleases, and RNases, with a control circuit to manage magnetic forces.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If conventional PCR methods are used for pathogen detection, then detection capability is achieved, but the requirement for complex laboratory equipment and cyclical temperature control makes it unsuitable for resource-limited settings
Solution Approach 1:
The patent changes the temperature parameter from cyclical (PCR) to isothermal (constant temperature), enabling pathogen detection without complex thermal cycling equipment. The LAMP amplification process maintains a constant temperature of 60-65°C, eliminating the need for sophisticated temperature control systems while achieving sensitive pathogen detection in resource-limited settings
Solution Approach 2:
The patent replaces the mechanical/thermal cycling system of PCR with a chemical/biochemical isothermal amplification system. By using strand-displacing DNA polymerase and carefully designed primers, the system achieves nucleic acid amplification through biochemical reactions at constant temperature, substituting complex thermal mechanics with simpler biochemical processes
2Measurement precision
If LAMP amplification is performed without effective mixing, then the amplification process can proceed, but mixing efficiency is insufficient leading to reduced detection sensitivity
Solution Approach 1:
The patent applies mechanical vibration through a vibration generator that vibrates the assay cartridge at specific frequencies during the LAMP amplification process. This vibration enhances the mixing of reagents and amplification products within the test well, ensuring uniform distribution of reactants and improving detection sensitivity by preventing local concentration gradients that could reduce amplification efficiency
Solution Approach 2:
The patent introduces dynamic mixing by making the normally static reagents and sample mobile through vibration. The vibration generator creates dynamic motion within the test well, causing continuous movement and redistribution of the reaction mixture. This dynamic approach ensures thorough mixing without requiring complex mechanical stirrers or pumps, maintaining system simplicity while improving mixing efficiency
3Measurement precision
If inhibitors like lactoferrin, lysozyme, nucleases, and RNases are present in the sample, then sample processing can proceed, but these inhibitors reduce amplification efficiency and detection accuracy
Solution Approach 1:
The patent converts the harmful effect of inhibitors into a beneficial selection mechanism by using a plunger component that applies mechanical force to rupture the sample carrier. This force selectively releases the sample while leaving inhibitors trapped or degraded, effectively using the harmful presence of inhibitors as a marker for their removal through controlled mechanical disruption and filtration
Solution Approach 2:
The patent introduces a plunger component as an intermediary mechanical element between the sample carrier and the reaction system. This plunger applies controlled force to rupture the sample carrier and facilitates the separation of sample from inhibitors through a defined mechanical pathway, acting as a mediator that enables selective sample release while blocking or removing inhibitory substances before they can interfere with amplification
4Productivity
If magnetic forces are applied to enhance mixing, then mixing efficiency improves, but the requirement for additional control mechanisms increases device complexity
Solution Approach 1:
The patent employs magnetic beads that self-generate mixing action through their interaction with applied magnetic fields. The magnetic beads inherently convert magnetic energy into mechanical motion, creating self-propelled mixing without requiring external mechanical stirrers or complex control systems. The beads' magnetic properties enable them to automatically respond to field application and generate the necessary mixing forces through their own magnetic moment and interaction with the field gradient
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
Enhances pathogen detection sensitivity and efficiency in resource-limited settings by effectively mixing samples and inhibiting inhibitors, facilitating accurate amplification and detection using LAMP.
Implementation Method 1
The force applied is the result of one or more of a magnetic field generator, a vibration generator, a sonic generator, and physical movement
Implementation Method 2
the at least one mixing object comprises at least one magnetic bead and wherein the force is exerted by a first magnetic field generated by a first magnet
Implementation Method 3
The force applied is the result of one or more of a magnetic field generator, a vibration generator, a sonic generator, and physical movement
Implementation Method 4
The force applied is the result of one or more of a magnetic field generator, a vibration generator, a sonic generator, and physical movement
Data Source
AI summary
The present application is generally directed to systems, methods, and devices for diagnostics for sensing and/or identifying pathogens, genomic materials, proteins, and/or other small molecules or biomarkers, for example, using loop-mediated isothermal amplification (LAMP). Some implementations include additional improvements, such as improvements to sample and reagent mixing, sample deposition, and compensation of inhibitors in the sample. Also disclosed herein are nucleic acid primers for use in the sensitive and specific detection of pathogens in biological samples by LAMP, which may be performed in the devices disclosed herein. The biological samples may be derived from patients including humans, plants, food, soil, contaminated surfaces, or animals such as livestock.