Automated point-of-care devices for complex sample processing and methods of use thereof
Inventors
Pais, Andrea Maria Dominic • Pais, Rohan Joseph Alexander
Assignees
Publication Number
US-12151239-B2
Publication Date
2024-11-26
Expiration Date
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Abstract
The present invention provides methods and devices for simple, low power, automated processing of biological samples through multiple sample preparation and assay steps. The methods and devices described facilitate the point-of-care implementation of complex diagnostic assays in equipment-free, non-laboratory settings.
Core Innovation
The present invention provides methods and devices for simple, low power, automated processing of biological samples through multiple sample preparation and assay steps to facilitate point-of-care implementation of complex diagnostic assays in equipment-free, non-laboratory settings. In accordance with the present invention, a sample-to-answer microfluidic device, an assay automation instrument, and a method for performing automated assays such as a nucleic acid amplification test (NAAT) on the microfluidic device are disclosed, comprising a portable assay automation instrument and a microfluidic cartridge containing stored reagents in liquid and dried format that are dispensed in a predefined sequence to perform a sample-to-answer NAAT.
The disclosure describes a reagent dispensing unit (RDU) that uses one or more reagent pouches containing aqueous reagents and a non-aqueous immiscible reagent packaged together with a frangible sealing layer, and one or more plungers and optional sharp objects or protrusions to rupture the frangible seal and deliver reagents to fluidic wells. The immiscible non-aqueous reagent is described as functioning to push out aqueous reagent to eliminate dead volume, form a barrier to prevent evaporation during heating steps, and reduce the need for complex valves, pumps, or precision metering of reaction chamber volumes.
The invention further teaches a rotatable microfluidic cartridge positioned between actuator elements comprising spatially oriented magnets and heater elements so that, in a single actuation step comprising rotation, magnetic beads can be captured, re-suspended, and transported between reagent wells to perform bind, wash, and elute sequences, and the amplification well can be thermally cycled by positioning over distinct heater elements. The disclosure also describes sample extraction and specimen processing units for maximizing sample elution from swabs and for sequential reagent delivery using rotary or wind-up actuators, flow-through reagent pouches to mitigate dead-volume losses, and lateral flow or other downstream detection units.
Claims Coverage
One independent claim is present with three main inventive features.
Reagent pouch comprising aqueous and non-aqueous immiscible reagent
A reagent-dispensing unit comprising at least one reagent pouch with a sealing layer, wherein the first non-compartmentalized reagent pouch comprises an aqueous reagent and a non-aqueous immiscible reagent packaged together in the first non-compartmentalized reagent pouch.
Actuator element with spatially oriented magnets and protrusion
At least one actuator element comprising spatially oriented magnets, and at least one protrusion configured to apply an actuation force to the at least one reagent pouch to dispense one or more reagents into a rotatable microfluidic cartridge; the actuator element may further comprise spatially oriented heater elements and protrusions to actuate flow-through pouches.
Rotatable microfluidic cartridge configured for magnetic bead capture and transport
A rotatable microfluidic cartridge comprising a plurality of wells, positioned proximate to the actuator element, configured to receive magnetic beads, and configured to rotate about a central axis such that in a single actuation step comprising rotating the microfluidic cartridge the spatially oriented magnets capture, re-suspend, and transport magnetic beads between the plurality of wells.
The independent claim centers on integration of mixed aqueous/immiscible reagent pouches, actuator elements with spatially oriented magnets/protrusions (and optional heaters), and a rotatable cartridge enabling magnetically driven capture, resuspension, and transport of magnetic beads among wells to achieve automated sample processing and assay steps.
Stated Advantages
Facilitates simple, low-power, automated processing of biological samples through multiple preparation and assay steps to enable point-of-care implementation in equipment-free, non-laboratory settings.
Eliminates dead volume issues and delivers aqueous reagents completely by using a non-aqueous immiscible fluid to push out aqueous reagent from reagent pouches and conduits, removing the need for sophisticated metering systems.
Reduces system complexity by avoiding the use of complex valves, pumps, and precision-molded metering chambers for accurate reagent delivery.
Provides an immiscible overlay that forms a barrier on top of the aqueous fluid to prevent evaporation during heating steps such as thermocycling or heat incubation.
Does not require precise actuation control for run-to-run repeatability because excess immiscible reagent overflows and ensures complete aqueous reagent delivery.
Enables rapid magnetic bead–based sample preparation including capture, re-suspension, and transport across wells, with an illustrative claim that the binding-to-elution sequence can be completed in two minutes.
Supports self-contained reagent storage and sequential reagent delivery to reduce hands-on time and operator-driven pipetting, lowering risk of user error and enabling use in low-resource and CLIA-waived settings.
Permits non-electrical actuation (e.g., wind-up spring) for automation without electric/battery power, advantageous in low-resource settings.
Provides an oil/immiscible phase dispensing system using a pressure head and venting to create smooth laminar oil flow and mitigate bubble formation without additional debubbling mechanisms.
Documented Applications
Sample-to-answer microfluidic device and assay automation instrument for performing nucleic acid amplification tests (NAATs), including PCR and isothermal amplification.
Point-of-care diagnostic assays in equipment-free, non-laboratory and low-resource settings, including efforts to enable CLIA-waived testing by minimizing sample preparation and fluid handling steps.
Magnetic bead–based sample preparation workflows on-chip to perform bind, wash, and elute sequences for nucleic acid purification prior to amplification.
Integrated detection using lateral flow strips for colorimetric read-out of amplified products and potential integration with fluorescence, electrochemical, or colorimetric detection technologies.
Sample collection and extraction devices for maximizing elution from swabs and sequential reagent delivery from a specimen processing unit with rotary or wind-up actuators.
Flow-through reagent pouch approaches for transferring and reconstituting reagents (including liquid magnetic bead suspensions or lyophilized reagents) into reaction chambers while mitigating dead-volume losses.
Downstream analysis units including DNA hybridization microarrays, protein arrays, and immunoassays as explicitly mentioned possible integrations.
Processing of a variety of biological sample types including saliva, blood, plasma, serum, urine, sputum, CSF, tissue, feces, plant, food, soil, and small organisms as examples of adaptable sample types.
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