Self-contained nucleic acid processing
Inventors
Johnson, Kyle K. • Brimhall, Owen D. • Politis, Jonathan J.
Assignees
Publication Number
US-11090652-B2
Publication Date
2021-08-17
Expiration Date
2037-07-28
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Abstract
Instruments and methods for amplifying nucleic acids in a sample provided in a flexible, self-contained, substantially closed sample container.
Core Innovation
The invention provides instruments and methods for amplifying nucleic acids in a sample using a flexible, self-contained, substantially closed sample container. The sample container features a loading attachment with sample and reagent zones, connected by sealable flow paths that allow input of sample and rehydration solution. Once loaded and sealed, the container is fully closed, minimizing contamination risk and ensuring containment.
A key element of the invention is the incorporation of a bead milling component, which operates with reciprocating paddle elements to generate a lysate from the sample by mechanically lysing cells within a designated lysis zone. The system further integrates compression members and seal members that enable directional movement of fluids between the fluidly connected reaction zones, facilitating process automation from sample lysis through nucleic acid extraction, purification, and amplification.
The invention targets challenges in molecular diagnostics relating to rapid, multiplex detection of infectious agents, overcoming issues of low pathogen concentration, sample volume limitations, and contamination through open handling. By automating difficult sample preparation steps, including adjustable lysis protocols and seamless transfer to amplification and detection steps within a fully closed container-hardware system, the invention improves robustness, reduces manual operations, and enhances diagnostic accuracy and speed.
Claims Coverage
The independent claims define several inventive features relating to self-contained nucleic acid amplification instruments, fluid handling, and bead milling mechanisms within flexible reaction containers.
Self-contained nucleic acid processing instrument with flexible, closed reaction container
The instrument is configured with an opening designed to receive a flexible sample container between two support members. The flexible container contains a fluid sample and multiple interconnected reaction zones: a lysis zone, a downstream nucleic acid extraction zone, and a downstream nucleic acid amplification zone. One or more sealable ports allow controlled fluidic access. The instrument is constructed so that, once the ports are sealed, the reaction container is fully closed, preventing external contamination.
Bead milling component with reciprocating paddle elements
A bead milling component, associated with one of the support members, comprises a reciprocating drive member operatively coupled to at least two adjacent (or side-by-side) paddle elements. These paddles are designed to extend alternately and repeatedly from a position adjacent to the support member outwards to contact the lysis zone, thus mechanically agitating the lysis zone and generating a lysate from the sample. The operation causes disruption of cellular material for downstream processing.
Integrated heater for temperature control in amplification zone
The instrument incorporates a heater positioned on a support member, specifically for controlling the temperature of nucleic acids within the amplification zone. This facilitates thermal cycling and precise temperature regulation necessary for efficient nucleic acid amplification (e.g., PCR).
The claims cover a self-contained nucleic acid analysis system featuring a closed, flexible reaction vessel, advanced fluid and sample handling using bead milling with paddle actuation, and integrated thermal control for automated, contamination-resistant sample processing and nucleic acid amplification.
Stated Advantages
Reduces contamination risk by enclosing all processes within a self-contained, fully closed sample container.
Automates complex sample preparation, including robust cell lysis and nucleic acid recovery, minimizing manual operations and error.
Enables efficient, high-sensitivity multiplex nucleic acid diagnostics from various sample types, even with low target concentrations or limited sample volumes.
Integrates all steps—from lysis through amplification and detection—within a single, secure platform for improved workflow and diagnostic speed.
Provides adjustable bead milling protocols to accommodate a wide variety of sample types, balancing effective lysis with nucleic acid integrity.
Documented Applications
Rapid extraction, processing, and identification of microbes or nucleic acids from biological, clinical, environmental, or veterinary samples.
Automated syndromic panel testing for infectious disease diagnosis, including high-throughput identification of multiple pathogens (viruses, bacteria, eukaryotes) in a single assay.
Analysis of tough-to-lyse samples such as spores, environmental particulates, or paraffin-embedded materials for nucleic acid-based testing.
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