Assay devices and methods of use thereof
Inventors
Ririe, Kirk M. • Wernerehl, Aaron D. • Pasko, Christopher P. • Laayoun, Ali • Vachon, Carole • Dupont-Filliard, Agnès • Mesta, Laurent • Hatch, Andrew C. • Huynh, Erik W. • Jones, David E.
Assignees
Biomerieux SA • Biofire Defense LLC • Biofire Diagnostics LLC
Publication Number
US-12172169-B2
Publication Date
2024-12-24
Expiration Date
2038-05-23
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Abstract
Systems, methods, and apparatuses are provided for self-contained nucleic acid preparation, amplification, and analysis.
Core Innovation
Described are self-contained reaction vessels, systems, and methods for rapid nucleic acid preparation, amplification, and analysis. The innovation uses a multi-zone vessel (also referred to as a pouch or card) that incorporates a sample lysis zone, a first-stage reaction zone, and a second-stage reaction zone comprising multiple reaction wells. These reaction vessels include all necessary reagents and components for sample processing, nucleic acid amplification, and analysis, allowing operation with minimal user intervention after sample introduction.
The invention addresses challenges in infectious disease diagnostics, including delayed diagnosis due to slow traditional methods and limitations of PCR workflows such as low pathogen concentrations and the impracticality of running large multiplex panels. Existing approaches are hampered by robustness, analysis complexity, contamination risk, and difficulty in processing varying sample types. The self-contained system aims to eliminate or minimize contamination, enable automation or semi-automation, allow simple packaging and manufacturing, and accelerate sample-to-answer workflows.
Key features include fluidly connected zones for sample preparation, nucleic acid recovery, and sequential amplification (first-stage and second-stage), with second-stage reaction wells preloaded with primers for parallel or differentiated analyte testing. The reaction vessel supports fabrication by automated or semi-automated methods, uses integrated reagent blisters or packs for both liquid and dry reagents, and provides flexible/rigid combinations for zones as needed. Fluid movement, thermal cycling, and reagent delivery are managed within the closed vessel, which can be operated in compatible instruments. The vessel enables simultaneous or sequential amplification and analysis with reduced user intervention and risk of contamination.
Claims Coverage
The patent claims cover one main inventive feature based on a single independent claim outlining the structure and function of a self-contained reaction vessel for nucleic acid analysis.
Self-contained reaction vessel with multi-zone flexible barrier structure and integrated reagent blisters
A self-contained reaction vessel designed for nucleic acid analysis comprising: - A sample lysis zone configured to lyse cells or spores present in a sample. - A first reaction zone fluidly connected to the sample lysis zone, which is configured for recovering nucleic acids from a lysed sample and for first-stage amplification of those nucleic acids. - A second-stage reaction zone fluidly connected (but separate from) the first reaction zone, the second-stage reaction zone containing multiple second-stage reaction wells. Each well includes a pair of primers, allowing further amplification, and all wells can be thermally cycled contemporaneously. - Multiple liquid reagent blisters, each fluidly connected to one or more of the sample lysis zone, the first reaction zone, or the second-stage reaction zone, with liquid reagents provided at the time of manufacture. - The vessel is constructed by at least a first flexible barrier film layer bonded to a second flexible barrier film layer, creating open spaces for each of the zones and blisters between the layers. The barrier film layers have a specified water vapor transmission rate (WVTR) in the range of about 0.05 g/m²/24 hrs to about 2 g/m²/24 hrs.
The patent provides coverage for a closed, multi-zone, flexible reaction vessel for self-contained nucleic acid sample preparation, amplification, and analysis, integrating reagent storage and fluidic connections, suitable for automated workflows.
Stated Advantages
Enables rapid, self-contained nucleic acid preparation, amplification, and analysis with minimal user input after sample introduction.
Reduces contamination risk by performing all steps within a closed system.
Supports automated or semi-automated manufacturing and operation for test devices, improving cost-effectiveness and simplicity.
Accelerates sample-to-answer time by integrating sample lysis, nucleic acid recovery, and amplification in a unified device.
Accommodates a variety of sample types and enables precise reagent delivery through onboard reagent blisters filled at manufacture.
Permits use with simple packaging, maximizes reagent stability with flexible barrier films, and can include both rigid and flexible zones as needed.
Documented Applications
Diagnosis of infectious diseases through rapid detection and identification of pathogens in clinical specimens.
Sample-to-answer molecular testing workflows in clinical laboratories and potentially point-of-care or field settings.
Amplification and analysis of nucleic acids from human, veterinary, industrial, or environmental samples, including but not limited to blood, urine, stool, saliva, soil, and water.
Assays for multiple pathogens and analytes via multiplex amplification and analysis in parallel reaction wells.
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