Multiplexed diagnostic detection apparatus and methods
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Publication Number
US-11453906-B2
Publication Date
2022-09-27
Expiration Date
Abstract
Disclosed herein are devices configured for the amplification and detection of multiple targets from a sample, and methods of using the same. The devices disclosed herein comprise microfluidic cartridges have a first stage (amplification) and a second (detection) stage. The two-stage design of the cartridges enables testing for multiple targets within a sample, i.e., from a single nucleic acid amplification reaction. Methods for the amplification and detection of a plurality of target nucleic acids from a sample are also disclosed herein.
Core Innovation
A multiplexed microfluidic cartridge includes a plurality of sample lanes, where each lane comprises a microfluidic network having an inlet and a single amplification chamber configured to be heated to amplify polynucleotides in a sample. The microfluidic network further includes a first amplification valve upstream of the single amplification chamber and a second amplification valve downstream of the single amplification chamber.
A plurality of detection chambers are disposed downstream of the single amplification chamber, with fluid communication between stages via microfluidic channels. The cartridge controls fluid transfer to allow filling multiple detection chambers from the single amplification chamber, with a first detection valve downstream of the single amplification chamber and upstream of the plurality of detection chambers, wherein the first detection valve is spatially separated from the second amplification valve.
A second detection valve downstream of the plurality of detection chambers isolates the sample within the detection chambers. The microfluidic channeling includes a second channel configured to transmit sample from the single amplification chamber via the first detection valve to two or more subchannels, each connecting to one of the plurality of detection chambers, and each detection chamber receives a divisible portion of the sample from the one isolatable amplification chamber.
During filling of two or more downstream detection chambers, the second amplification valve is closed when the sample is transmitted from the single amplification chamber to the plurality of detection chambers downstream of the single amplification chamber. A valve isolating the one isolatable amplification chamber from the microfluidic channel is closed when the sample is propelled downstream with a motive force applied to the sample.
Claims Coverage
The independent claims are clm-00001, clm-00009, and clm-00015, each directed to a microfluidic cartridge architecture for amplified polynucleotides transferred to multiple downstream detection chambers using isolation valves between amplification and detection stages. The main inventive features center on a single or one isolatable amplification chamber, separated upstream and downstream valves for stage isolation, and microfluidic channeling that enables distributing a sample into two or more detection chambers while closing the amplification stage during transfer.
Single heated amplification chamber with separated upstream and downstream amplification valves
A microfluidic cartridge having a plurality of sample lanes, each lane including a single amplification chamber configured to be heated to amplify polynucleotides, with a first amplification valve upstream of the single amplification chamber and a second amplification valve downstream of the single amplification chamber.
Multiple downstream detection chambers isolated by spatially separated detection valves
A plurality of detection chambers downstream of the single amplification chamber, with a first detection valve downstream of the single amplification chamber and upstream of the plurality of detection chambers, wherein the first detection valve is spatially separated from the second amplification valve, and a second detection valve downstream of the plurality of detection chambers isolating the sample within the plurality of detection chambers.
Distributed filling of two or more detection chambers via subchannels while closing the amplification valve
A second channel transmitting the sample from the single amplification chamber via the first detection valve to two or more subchannels, each subchannel connecting to one of the plurality of detection chambers to allow filling of two or more detection chambers, wherein the second amplification valve is closed when the sample is transmitted to the plurality of detection chambers.
Multiplexed divisibility into multiple isolatable detection chambers with motive force propelling
A multiplexed microfluidic cartridge having one isolatable amplification chamber configured to be heated to amplify polynucleotides, and a plurality of isolatable detection chambers downstream, wherein each detection chamber receives a divisible portion of the sample from the one isolatable amplification chamber, and a microfluidic channel configured to allow filling of two or more detection chambers with a motive force applied to the sample to propel the sample, with a valve isolating the one isolatable amplification chamber from the microfluidic channel closed when the sample is propelled downstream.
Across the independent claims, the central claimed concept is a multiplexed microfluidic cartridge architecture that amplifies polynucleotides in a single or one isolatable heated amplification chamber, then isolates and transfers sample to multiple downstream detection chambers using spatially separated valves and microfluidic channeling that distributes a divisible portion into two or more detection chambers while closing the amplification stage during that downstream filling step.
Stated Advantages
Documented Applications
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