Integrated apparatus for performing nucleic acid extraction and diagnostic testing on multiple biological samples

• US 9,259,734 B2
• Filed: 03/09/2012
• Issued: 02/16/2016
• Est. Priority Date: 07/13/2007
• Status: Active Grant

• Current Assignee: HandyLab Incorporated (Becton, Dickinson & Co)
• Original Assignee: HandyLab Incorporated (Becton, Dickinson & Co)
• Inventors: Williams, Jeff, Handique, Kalyan, Wilson, Kerry
• Primary Examiner(s): Bowers, Nathan
• Assistant Examiner(s): Prakash, Gautam
• Application Number:
  US13/417,108
• Publication Number:
  US 20120171759A1
• Time in Patent Office: 1,439 Days
• Field of Search: 435/287.2, 435/6.1, 435/6.11, 435/6.12
• 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 3/5027 : 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 3/5085 : for multiple samples, e.g. microtitration plates
• 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 9/06 : Test-tube stands; Test-tube holders
• CPC Class Codes: B01L 9/527 : for microfluidic devices, e.g. used for lab-on-a-chip
• 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 2035/00881 : network configurations
• CPC Class Codes: G01N 2035/0436 : with pre-packaged reagents, i.e. test-packs
• CPC Class Codes: G01N 2035/103 : using disposable tips
• CPC Class Codes: G01N 35/026 : having blocks or racks of reaction cells or cuvettes

Abstract

The technology described herein generally relates to systems for extracting polynucleotides from multiple samples, particularly from biological samples, and additionally to systems that subsequently amplify and detect the extracted polynucleotides. The technology more particularly relates to microfluidic systems that carry out PCR on multiple samples of nucleotides of interest within microfluidic channels, and detect those nucleotides.

Claims

1. An apparatus configured to extract and amplify nucleic acid from a plurality of nucleic-acid containing samples, comprising:

a rack comprising a plurality of lanes, each lane of the plurality of lanes comprising

a first location configured to receive one of the plurality of samples, and

a second location configured to receive a reagent holder;

an integrated separator and heater comprising

a magnetic separator configured to move adjacent to one or more of the second locations of the rack; and

a heater assembly comprising a plurality of independently controllable heater units, each heater unit shaped and sized to accept a single process chamber of a reagent holder, each heater unit configured to apply heat independently of any other heater unit in the heater assembly;

a liquid dispenser movable from a first position above one of the first locations to a second position above one of the second locations; and

a bay comprising at least one heat source and configured to receive a microfluidic cartridge comprising a plurality of lanes, the bay configured to interact with the cartridge to separately amplify nucleic acid extracted from the plurality of samples by applying heat to each lane individually.

2. The apparatus of claim 1, further comprising a reagent holder comprising a process chamber and one or more reagent receptacles received in each of the second locations.

3. The apparatus of claim 2, wherein the magnetic separator is configured to apply a magnetic force to contents of the process chamber of each holder.

4. The apparatus of claim 2, further comprising a processor configured to control heating functions of the heater assembly in conjunction with operation of the magnetic separator.

5. The apparatus of claim 1, wherein the liquid dispenser is movable from the second position above one of the second locations to a third position above the bay.

6. The apparatus of claim 1, further comprising a detection system comprising a light source configured to emit light in an absorption band of a fluorescent dye and a light detector configured to detect light in an emission band of the fluorescent dye.

7. The apparatus of claim 1, wherein the at least one heat source is configured to be thermally coupled to the microfluidic cartridge and configured to apply heat to one or more selected regions of the microfluidic cartridge at one or more selected times, in order to

apply thermocycling operations sufficient to amplify the extracted nucleic acid.

8. The apparatus of claim 7, further comprising a processor configured to control applying heat to the one or more selected regions of the microfluidic cartridge at one or more selected times.

9. The apparatus of claim 1, wherein the at least one heat source comprises at least two contact heat sources, wherein the at least two contact heat sources are each configured to be independently thermally coupled to a different selected region of the microfluidic cartridge, whereby the different selected regions are independently heated.

10. The apparatus of claim 1, further comprising the microfluidic cartridge, wherein the microfluidic cartridge is configured to amplify nucleic acid by a method selected from the group consisting of:

PCR;

TMA;

SDA;

NASBA;

LCR; and

Rolling-Cycle Amplifications.

11. The apparatus of claim 1, wherein the apparatus is integrated into a single housing.

12. The apparatus of claim 1, wherein the plurality of samples are each independently selected from the group consisting of:

saliva, urine, blood, semen, mucus, and spinal fluid.

13. The apparatus of claim 1, wherein the second location is configured to receive a removable reagent holder, each second location allowing removal of a reagent holder independent of any other second location.

14. A diagnostic apparatus, comprising:

one or more racks, each configured to receive a number of samples and a corresponding number of holders, each holder configured to receive a process chamber and one or more receptacles configured to store reagents;

an integrated separator and heater comprising

a magnetic separator configured to move adjacent to the process chambers of a plurality of holders; and

a heater assembly comprising a number of independently controllable heater units, each heater unit shaped and sized to accept one process chamber, each heater unit configured to apply heat independently of any other heater unit in the heater assembly;

a liquid dispenser movable from a first position above a first holder to a second position above a second holder, and movable from the first position above the first holder to a different position above the first holder; and

one or more bays, each bay having a shape complementary to a shape of a microfluidic cartridge having a plurality of channels, wherein each bay is configured to cooperate with the microfluidic cartridge to separately process a material extracted from the number of samples by applying heat to each channel individually.

15. The apparatus of claim 14, further comprising instructions for controlling heating functions of the heater assembly in conjunction with operation of the magnetic separator.

16. The apparatus of claim 14, wherein the material is nucleic acid, and wherein the apparatus further comprises the microfluidic cartridge received in one or more of the bays, the microfluidic cartridge comprising a number of inlets each of which is in fluid communication with one of the channels, wherein the microfluidic cartridge is configured to amplify the nucleic acid extracted from one of the number of samples.

17. The apparatus of claim 16, wherein the microfluidic cartridge further comprises one or more windows that permit detection of amplified nucleic acid, wherein the apparatus further comprising one or more detection systems positioned in proximity to the one or more windows.

18. The apparatus of claim 16, wherein the liquid dispenser is movable from a position above one of the holders to a position above one of the number of inlets.

19. The apparatus of claim 14, wherein the liquid dispenser is movable from a position above a holder to a position above the one or more bays.

20. The apparatus of claim 14, wherein the one or more bays comprise at least one heat source configured to be thermally coupled to the microfluidic cartridge and configured to apply heat to one or more selected regions of the microfluidic cartridge at one or more selected times, in order to

apply a predetermined thermal program to the material.

21. The apparatus of claim 20, wherein the at least one heat source comprises at least two contact heat sources, wherein the at least two contact heat sources are each configured to be independently thermally coupled to a different selected region of the microfluidic cartridge, whereby the different selected regions are independently heated.

22. The apparatus of claim 14, wherein the one or more racks are each configured to receive a number of samples and a corresponding number of removable holders, the one or more racks allowing removal of each holder independent of any other holder.