Integrated system for processing microfluidic samples, and method of using same
Inventors
Handique, Kalyan • Brahmasandra, Sundaresh N. • Ganesan, Karthik • Wu, Betty • Phadke, Nikhil • Parunak, Gene • Williams, Jeff
Assignees
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Publication Number
US-11666903-B2
Publication Date
2023-06-06
Expiration Date
Abstract
This patent application describes an integrated apparatus for processing polynucleotide-containing samples, and for providing a diagnostic result thereon. The apparatus is configured to receive a microfluidic cartridge that contains reagents and a network for processing a sample. Also described are methods of using the apparatus.
Core Innovation
The disclosed invention provides an apparatus for processing and amplifying a plurality of polynucleotide-containing samples using an automated sampling device, a reagent reservoir, and a disposable multi-lane microfluidic cartridge supported on a platform. The multi-lane microfluidic cartridge includes a plurality of sample lanes, with each lane having a dedicated pipette inlet and a region where amplification occurs. The cartridge is configured to permit loading of polynucleotide-containing samples at different times, such that a first sample is passed within the microfluidic cartridge at a different time than a second sample.
A first heat source, separate from the microfluidic cartridge, applies heat to one or more selected regions of the microfluidic cartridge at one or more selected times to prepare one or more polynucleotides from the polynucleotide-containing samples for amplification. Multiple second heat sources, also separate from the microfluidic cartridge, align with selected regions and apply heat at selected times to amplify the prepared polynucleotides in an automated manner. In this approach, amplification includes cyclically heating in a series of heating phases after the cartridge is supported by the platform, where each heating phase cycles between at least two temperatures and maintains a substantially uniform temperature within the amplification region at each temperature.
The apparatus further integrates cartridge processing and detection functions by including, within a sample lane, a processing chamber that is heated by the first heat source. A magnet moves into and out of place to apply a magnetic field to a polynucleotide-loaded retention member in the processing chamber when the retention member is received in the cartridge. An optical detector detects the presence of the one or more amplified polynucleotides within the region where amplification occurs.
Claims Coverage
The independent claim defines an integrated, automated apparatus with multi-lane microfluidic processing, separate heat sources for preparing and amplifying polynucleotides, cyclic heating with substantially uniform temperature control, magnetic-field handling of a polynucleotide-loaded retention member, and optical detection of amplified polynucleotides. The inventive features below consolidate the overlapping independent claim content across the input items.
Automated multi-lane cartridge processing with time-different sample loading
A platform supports a multi-lane microfluidic cartridge having a plurality of sample lanes, each with a dedicated pipette inlet and a region where amplification occurs, and the cartridge permits loading polynucleotide-containing samples at different times so that a first sample is passed at a different time than a second sample.
First separate heat source for polynucleotide preparation
A first heat source separate from the multi-lane microfluidic cartridge applies heat to one or more selected regions at one or more selected times to prepare one or more polynucleotides from the polynucleotide-containing samples for amplification.
Second separate heat sources for automated amplification in selected regions
A plurality of second heat sources separate from the multi-lane microfluidic cartridge align with selected regions and apply heat at selected times to amplify the prepared polynucleotides in an automated manner.
Cyclic multi-temperature amplification phases with substantially uniform temperature
A second heat source cyclically heats in a series of heating phases, each phase cycling between at least two temperatures and maintaining a substantially uniform temperature within the region where amplification occurs at each temperature.
Magnetic-field actuation of polynucleotide-loaded retention member
A magnet moves into and out of place to apply a magnetic field to a polynucleotide-loaded retention member in a processing chamber when the cartridge is supported by the platform.
Optical detection of amplified polynucleotides within amplification region
An optical detector detects the presence of one or more amplified polynucleotides within the region where amplification occurs.
Overall, the claim coverage centers on automated processing and amplification of multiple polynucleotide-containing samples in a multi-lane microfluidic cartridge, with separate heat sources for preparation and amplification, cyclic heating between at least two temperatures with substantially uniform temperature in the amplification region, magnetic-field handling of a polynucleotide-loaded retention member, and optical detection of amplified polynucleotides.
Stated Advantages
Automated amplification reactions performed in an automated manner.
Cyclical heating in series of heating phases with cyclic heating between at least two temperatures.
Substantially uniform temperature within the region where amplification occurs at each temperature.
Optical detection of amplified polynucleotides within the region where amplification occurs.
Enable reduction of PCR inhibitors enabling PCR-ready samples without centrifugation.
Self-contained reagent storage through sealed/actuated reagent reservoirs.
On-demand portability.
Automated multi-sample processing using a cartridge platform approach, including multiplexed analysis across multiple cartridges.
Documented Applications
Processing and amplifying multiple polynucleotide-containing samples using a bench-top real-time PCR device and a microfluidic PCR operation with automated amplification in a multi-lane microfluidic cartridge supported on a platform.
Genotyping/assay workflow targeting GBS (Group B Streptococcus), including detection concepts described for GBS (e.g., cfb gene, primers/probes, and internal controls) in lyophilized PCR pellet implementations.
Automated processing and amplification of a plurality of polynucleotide-containing samples using a multi-lane microfluidic cartridge platform for diagnostic use.
PCR and optical fluorescence detection of a plurality of polynucleotide-containing samples.
Multiplexed analysis across multiple cartridges, including concurrent or successive processing of multiple polynucleotide-containing samples loaded at different times.
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