Microfluidic system for amplifying and detecting polynucleotides in parallel

• US 9,080,207 B2
• Filed: 12/03/2012
• Issued: 07/14/2015
• Est. Priority Date: 03/24/2006
• Status: Active Grant

• Current Assignee: HandyLab Incorporated (Becton, Dickinson & Co)
• Original Assignee: HandyLab Incorporated (Becton, Dickinson & Co)
• Inventors: Handique, Kalyan, Brahmasandra, Sundaresh N, Ganesan, Karthik, Williams, Jeff
• Primary Examiner(s): Kim, Young J
• Application Number:
  US13/692,929
• Publication Number:
  US 20130101990A1
• Time in Patent Office: 953 Days
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• 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 2200/147 Employing temperature sensors
• CPC Class Codes: B01L 2200/148 Specific details about calibrations
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/021 Identification, e.g. bar codes
• CPC Class Codes: B01L 2300/045 whereby the whole cover is slidable
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/1861 using radiation
• CPC Class Codes: B01L 2400/0442 thermal energy, e.g. vaporisation, bubble jet
• CPC Class Codes: B01L 2400/0481 squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0611 duck bill valves
• CPC Class Codes: B01L 2400/0677 phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
• CPC Class Codes: B01L 2400/0683 : mechanically breaking a wall or membrane within a channel or chamber
• CPC Class Codes: B01L 3/502715 : characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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: B01L 9/527 : for microfluidic devices, e.g. used for lab-on-a-chip
• 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: C12Q 1/686 : Polymerase chain reaction [PCR]
• CPC Class Codes: F16K 2099/0084 : Chemistry or biology, e.g. "lab-on-a-chip" technology
• CPC Class Codes: F16K 99/0001 : Microvalves microdevices B81B1/00; manufacture or treatment of devices or systems in or on a substrate B81C1/00; microfluidic structures B01L3/5027; micropumps F04B19/006
• CPC Class Codes: F16K 99/003 : Valves for single use only
• CPC Class Codes: F16K 99/0032 : using phase transition or influencing viscosity
• CPC Class Codes: F16K 99/0044 : using thermo-electric means
• CPC Class Codes: F16K 99/0061 : actuated by an expanding gas or liquid volume
• CPC Class Codes: G01N 2021/6419 : Excitation at two or more wavelengths
• CPC Class Codes: G01N 2021/6421 : Measuring at two or more wavelengths
• CPC Class Codes: G01N 2021/6441 : with two or more labels
• CPC Class Codes: G01N 2035/00881 : network configurations
• CPC Class Codes: G01N 21/6428 : Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53

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.

Claims

1. An apparatus, comprising:

a plurality of multi-lane microfluidic cartridges, each lane comprising a reaction zone;

a plurality of receiving bays, each receiving bay configured to receive one of the plurality of microfluidic cartridges;

each reaction zone comprising multiple separately controllable heaters thermally coupled thereto, the heaters configured to simultaneously and uniformly heat the reaction zone to carry out nucleic acid amplification on a polynucleotide-containing sample in the reaction zone;

a detector configured to detect the presence of an amplification product in one or more reaction zones; and

a processor coupled to the detector and the heaters, configured to control heating of one or more reaction zones by the heaters.

2. The apparatus of claim 1, wherein the separately controllable heaters thermally coupled to each reaction zone are configured to maintain a temperature gradient of less than 1°

C. across a width of the reaction zone at any point along a length of the reaction zone.

3. The apparatus of claim 1, wherein the processor is programmable to operate the detector to detect a polynucleotide or a probe thereof in a plurality of microfluidic cartridges located in the plurality of receiving bays.

4. The apparatus of claim 1, wherein the detector comprises an optical detector, the optical detector comprising a light source that selectively emits light in an absorption band of a fluorescent dye and a light detector that selectively detects light in an emission band of the fluorescent dye, wherein the flourescent dye is used to detect a nucleic acid amplification product.

5. The apparatus of claim 4, wherein the optical detector comprises a bandpass-filtered diode that selectively emits light in the absorption band of the fluorescent dye and a bandpass filtered photodiode that selectively detects light in the emission band of the fluorescent dye.

6. The apparatus of claim 1, further comprising instructions that direct the processor to maintain a substantially uniform temperature throughout a reaction chamber thermally coupled to the heaters.

20. A method of carrying out nucleic acid amplification on a plurality of samples, the method comprising:

introducing the plurality of samples into the plurality of multi-lane microfluidic cartridges of claim 1, wherein each lane comprises a reaction zone configured to permit heating of a sample independently of the other samples;

moving the plurality of samples into the respective plurality of reaction zones; and

heating the plurality of reaction zones in order to amplify polynucleotides contained with the plurality of samples in the plurality of multi-lane microfluidic cartridges, at least one reaction zone separately thermally controllable from another reaction zone.

21. The method of claim 20, further comprising detecting the presence of a polynucleotide or a polynucleotide probe in the plurality of samples.

22. The method of claim 20, wherein heating the reaction zones comprises maintaining a substantially uniform temperature throughout each reaction zone.

7. A device for carrying out nucleic acid amplification on a plurality of samples, the device comprising:

a plurality of multi-lane microfluidic cartridges, each lane comprising a reaction zone;

a plurality of receiving bays, each receiving bay configured to receive one of the plurality of microfluidic cartridges;

a plurality of separately controllable heaters thermally coupled to each reaction zone, the heaters configured to uniformly heat the reaction zone to carry out nucleic acid amplification on a polynucleotide-containing sample in the reaction zone;

a detector configured to detect the presence of an amplification product in one or more reaction zones;

a processor coupled to the detector and the plurality of separately controllable heaters, configured to control heating of one or more reaction zones by the plurality of separately controllable heaters; and

an input device coupled to the processor and configured to permit concurrent or consecutive control of the plurality of multi-lane microfluidic cartridges.

8. The device of claim 7, wherein at least one of the plurality of separately controllable heaters is a first contact heat source selected from a resistive heater, a radiator, a fluidic heat exchanger, and a Peltier device.

9. The device of claim 8, wherein the first contact heat source is thermally coupled to a distinct location in a first microfluidic cartridge received in a first receiving bay, whereby the distinct location is selectively heated.

10. The device of claim 9, wherein the distinct location has a surface area of between about 1 mm² and about 225 mm².

11. The device of claim 8, further comprising a second contact heat source configured to be independently thermally coupled to a distinct location in a second microfluidic cartridge received in a second receiving bay, whereby the distinct location in the second microfluidic cartridge is independently heated from the distinct location in the first microfluidic cartridge.

12. The device of claim 8, wherein the first contact heat source is configured to be in direct physical contact with the distinct location of the first microfluidic cartridge received in the first receiving bay.

13. The device of claim 8, further comprising a compliant layer configured to thermally couple the first contact heat source with at least a portion of the first microfluidic cartridge received in the first receiving bay.

14. The device of claim 7, wherein at least one of the plurality of separately controllable heaters is a radiative heat source configured to direct heat to a distinct location of a first microfluidic cartridge received in a first receiving bay.

15. The device of claim 7, wherein the input device is selected from the group consisting of a keyboard, a touch-sensitive surface, a microphone, a hard disk drive, an optical disk drive, a serial connection, a parallel connection, a wireless network connection, a wired network connection, and a mouse.

16. The device of claim 7, further comprising at least one sample identifier coupled to the processor, the sample identifier being selected from an optical character reader, a bar code reader, and a radio frequency tag reader.

17. The device of claim 7, further comprising at least one output coupled to the processor, the output being selected from a display, a printer, a speaker, a serial connection, a parallel connection, a wireless network connection, and a wired network connection.

18. The device of claim 7, further comprising a heating stage configured to be removable from the device, wherein at least one of the plurality of separately controllable heaters is located in the heating stage.

19. The device of claim 7, further comprising instructions that direct the processor to maintain a substantially uniform temperature throughout a reaction chamber thermally coupled to the heaters.