Microfluidic system for amplifying and detecting polynucleotides in parallel
Inventors
Handique, Kalyan • Brahmasandra, Sundaresh N. • Ganesan, Karthik • Williams, Jeff
Assignees
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Publication Number
US-11142785-B2
Publication Date
2021-10-12
Expiration Date
Abstract
The present technology provides for an apparatus for detecting polynucleotides in samples, particularly from biological samples. The technology more particularly relates to microfluidic systems that carry out PCR on nucleotides of interest within microfluidic channels, and detect those nucleotides. The apparatus includes a microfluidic cartridge that is configured to accept a plurality of samples, and which can carry out PCR on each sample individually, or a group of, or all of the plurality of samples simultaneously.
Core Innovation
The invention provides a microfluidic PCR diagnostic system in which a multi-lane microfluidic cartridge includes an amplification region in each lane for amplification of polynucleotides. An instrument includes a receiving bay configured to receive the multi-lane microfluidic cartridge, and the instrument controls amplification and detects amplification products in the amplification region during amplification. The instrument includes separately controllable heat sources to direct heat to an amplification region of a lane when the cartridge is received in the receiving bay, together with a detector mounted in the instrument configured to detect an amplification product during amplification.
The system further incorporates an automatic pipetting machine mounted in the instrument that loads polynucleotide-containing samples into the multi-lane microfluidic cartridge when the cartridge is received in the receiving bay. The automatic pipetting machine is configured to add a polynucleotide-containing sample after a different polynucleotide-containing sample has been amplified, and the amplified polynucleotide-containing sample remains in the microfluidic cartridge while another polynucleotide-containing sample is added to an unused lane. The instrument is configured to move the multi-lane microfluidic cartridge from a first location inside the instrument to a second location before amplification is carried out.
The invention also supports detector-based detection of amplification products and multi-lane, multiplexed detection using a detector configured to independently detect a plurality of fluorescent dyes having different fluorescent emission spectra, where each fluorescent dye corresponds to a fluorescent polynucleotide probe or a fragment thereof. The instrument and cartridge support high throughput workflows, including processing successive samples in unused lanes while previously amplified samples remain, and refined configurations with a second receiving bay and a touchscreen display.
Claims Coverage
The provided independent claims cover four main inventive aspects: a multi-lane cartridge with lane-specific amplification regions; an instrument receiving bay with separately controllable heat sources; a detector for detecting amplification products, including multi-dye fluorescence in refined versions; and automatic pipetting that adds samples to unused lanes while prior amplified samples remain, with cartridge movement before amplification in one claim.
Multi-lane cartridge with lane amplification regions
Providing a multi-lane microfluidic cartridge, each lane comprising an amplification region within which amplification of polynucleotides is carried out.
Receiving bay with separately controllable heat sources for amplification
An instrument comprising a receiving bay configured to receive the multi-lane microfluidic cartridge and a plurality of separately controllable heat sources mounted in the instrument, with a heat source configured to direct heat to an amplification region of a lane when the cartridge is received in the receiving bay.
Detector detecting amplification products in amplification regions
A detector mounted in the instrument and configured to detect the presence of an amplification product in the amplification region during amplification.
Automatic pipetting of successive samples into unused lanes while amplified samples remain
An automatic pipetting machine configured to load polynucleotide-containing samples into the multi-lane microfluidic cartridge, add a polynucleotide-containing sample after a different polynucleotide-containing sample has been amplified, and leave the amplified polynucleotide-containing sample in the cartridge while another sample is added to an unused lane.
Cyclic heating phases for amplification
A plurality of separately controllable heat sources configured to cyclically heat in a series of heating phases to amplify polynucleotides in the amplification region, each heating phase comprising cycling between at least two temperatures.
Movement of cartridge within the instrument before amplification
The instrument is configured to move the multi-lane microfluidic cartridge from a first location inside the instrument to a second location before amplification of polynucleotides is carried out.
Independent multi-dye fluorescence detection
The detector is configured to independently detect a plurality of fluorescent dyes in the amplification region, the plurality of fluorescent dyes having different fluorescent emission spectra and each fluorescent dye corresponding to a fluorescent polynucleotide probe or a fragment thereof.
The coverage centers on a multi-lane microfluidic cartridge with lane amplification regions, an instrument receiving bay with separately controllable heat sources to drive amplification, a detector for detecting amplification products, and an automatic pipetting machine that adds successive polynucleotide-containing samples to unused lanes while amplified material remains in the cartridge. Refined coverage includes cyclic heating phases, independent detection of multiple fluorescent dyes with different emission spectra, and movement of the cartridge within the instrument before amplification.
Stated Advantages
Documented Applications
Not explicitly described in patent.
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