MULTIPLEXED DIAGNOSTIC DETECTION APPARATUS AND METHODS
First Claim
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1. A microfluidic cartridge comprising:
- a plurality of sample lanes, each lane comprising a microfluidic network having;
an inlet;
an amplification chamber in which the polynucleotides in a liquidic sample are amplified;
a first amplification valve upstream of the amplification chamber in an initial open position prior to introduction of a sample and a first amplification gale upstream of the amplification chamber in an initial closed position prior to introduction of a sample;
a second amplification valve downstream of the amplification chamber in an initial open position and a second amplification gate downstream of the amplification chamber in an initial closed position prior to introduction of a sample;
a first channel leading from the inlet, via the first amplification valve and first amplification gate, to the amplification chamber;
a plurality of detection chambers downstream of the second amplification gate;
a first detection valve upstream of the plurality of detection chambers and a second detection valve downstream of the plurality of detection chambers, the first and second detection valves each in an initial open position prior to introduction of a sample; and
a second channel leading from the amplification chamber, via the second amplification gate and first detection valve, to the plurality of detection chambers.
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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.
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Citations
68 Claims
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1. A microfluidic cartridge comprising:
a plurality of sample lanes, each lane comprising a microfluidic network having; an inlet; an amplification chamber in which the polynucleotides in a liquidic sample are amplified; a first amplification valve upstream of the amplification chamber in an initial open position prior to introduction of a sample and a first amplification gale upstream of the amplification chamber in an initial closed position prior to introduction of a sample; a second amplification valve downstream of the amplification chamber in an initial open position and a second amplification gate downstream of the amplification chamber in an initial closed position prior to introduction of a sample; a first channel leading from the inlet, via the first amplification valve and first amplification gate, to the amplification chamber; a plurality of detection chambers downstream of the second amplification gate; a first detection valve upstream of the plurality of detection chambers and a second detection valve downstream of the plurality of detection chambers, the first and second detection valves each in an initial open position prior to introduction of a sample; and a second channel leading from the amplification chamber, via the second amplification gate and first detection valve, to the plurality of detection chambers. - View Dependent Claims (2, 3, 9, 10, 11, 12, 14)
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4-8. -8. (canceled)
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13. (canceled)
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15-56. -56. (canceled)
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57. A microfluidic cartridge comprising:
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a plurality of sample lanes, each lane comprised of a microfluidic network having a first and a second stage connected by a microfluidic channel, the first stage comprising; an inlet; an amplification chamber in fluid communication with the inlet; a first amplification valve and a first amplification gate, both upstream of the amplification chamber; a second amplification valve and a second amplification gate, both downstream of the amplification chamber; a first vent; the second stage being distal to the first stage along the microfluidic network and comprising; a plurality of detection chambers; a first detection valve upstream of the plurality of detection chambers and a second detection valve downstream of the plurality of detection chambers; and a second vent. - View Dependent Claims (58, 59, 60, 61, 62)
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63. A multiplexed microfluidic cartridge comprising:
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a plurality of sample lanes, each lane comprised of a microfluidic network having; one isolatable amplification chamber in which the polynucleotides in a liquidic sample are amplified; a plurality of isolatable detection chambers downstream of the amplification chamber, each detection chamber in fluid communication with the amplification chamber such that each detection chamber receives a divisible portion of the amplified sample from the amplification chamber; wherein the amplification chamber and detection chambers are connected via a microfluidic channel, the microfluidic channel including a gate and valve between the amplification chamber and detection chambers, wherein the gate is in an initially closed position prior to insertion of a sample and the valve is in an initially open position prior to insertion of a sample. - View Dependent Claims (64, 65, 66, 67, 68)
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Specification