MULTIPLEXED DIAGNOSTIC DETECTION APPARATUS AND METHODS

• US 20140329301A1
• Filed: 05/01/2014
• Published: 11/06/2014
• Est. Priority Date: 11/04/2011
• Status: Active Grant

• Current Assignee: HandyLab Incorporated (Becton, Dickinson & Co)
• Original Assignee: HandyLab Incorporated (Becton, Dickinson & Co)
• Inventors: Handique, Kalyan
• Granted Patent: US 11,453,906 B2
• Time in Patent Office: Days
• US Class Current: 435/287.2
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2400/06 Valves, specific forms thereof
• CPC Class Codes: B01L 2400/0677 phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502723 characterised by venting arrangements
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: F16K 2099/0084 Chemistry or biology, e.g. "lab-on-a-chip" technology
• CPC Class Codes: F16K 99/0036 operated by temperature variations

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.

Claims

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.

2. The microfluidic cartridge of claim 1, further comprising a third channel in each of the microfluidic networks, the third channel leading from the amplification chamber, via the second amplification valve, to a first vent.

3. The microfluidic cartridge of claim 2, further comprising a fourth channel in each of the microfluidic networks, the fourth channel leading from the plurality of detection chambers, via the second detection valve to a second vent.

9. The microfluidic cartridge of claim 1, wherein the amplification valves comprise a temperature responsive substance that melts upon heating to seal one of at least the first and second channels in fluid communication with the amplification chamber.

10. The microfluidic cartridge of claim 1, wherein the detection valves comprise a temperature responsive substance that melts upon heating to seal one of at least the second and fourth channels that communicates with the plurality of detection chambers.

11. The microfluidic cartridge of claim 1, wherein the amplification chamber has a volume of 5-10 μ

l.

12. The microfluidic cartridge of claim 1, wherein the plurality of detection chambers each has a volume of approximately 1 μ

l.

14. The microfluidic cartridge of claim 1, further comprising one or more fluorescence detection windows disposed over the detection chambers.

4-8. -8. (canceled)

13. (canceled)

15-56. -56. (canceled)

57. A microfluidic cartridge comprising:

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.

58. The cartridge of claim 57, wherein the first and second amplification gates are in an initially closed position prior to introduction of a sample.

59. The cartridge of claim 57, wherein the first and second amplification valves are in an initially open position prior to introduction of a sample.

60. The cartridge of claim 57, wherein the first vent is downstream of the second amplification valve, but is not downstream of the second amplification gate.

61. The cartridge of claim 57, wherein the first and second stages of each microfluidic network are in the same horizontal plane.

62. The cartridge of claim 57, wherein the liquidic volume of the first stage is greater than the liquidic volume of the second stage.

63. A multiplexed microfluidic cartridge comprising:

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.

64. The cartridge of claim 63, wherein each microfluidic network comprises one amplification chamber and six detection chambers.

65. The cartridge of claim 63, wherein each of the plurality of detection chambers in the microfluidic network is of equal volume.

66. The microfluidic cartridge of claim 65, wherein each detection chamber has a volume of approximately 1 μ

l.

67. The microfluidic cartridge of claim 63, wherein the amplification chamber has a volume of approximately 5-10 μ

l.

68. The microfluidic cartridge of claim 63, wherein the collective volume of the detection chambers is less than the volume of the amplification chamber.