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

• US 9,186,677 B2
• Filed: 07/14/2008
• Issued: 11/17/2015
• 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, Wilson, Kerry, Handique, Kalyan
• Primary Examiner(s): Hyun, Paul
• Assistant Examiner(s): EOM, ROBERT J
• Application Number:
  US12/218,498
• Publication Number:
  US 20090221059A1
• Time in Patent Office: 2,682 Days
• Field of Search: 422/63, 422/65, 422/560, 422/561, 422/562, 422/564
• 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 2300/06 Auxiliary integrated devices, integrated components
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• 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 2300/0832 cylindrical, tube shaped
• 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 2300/18 Means for temperature control
• 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 3/0275 : Interchangeable or disposable dispensing tips
• 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 3/502761 : specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B01L 3/52 : Containers specially adapted for storing or dispensing a reagent B01L3/02 takes precedence; containers for medical or pharmaceutical purposes A61J1/00; containers in general B65D; storing or dispensing test elements G01N33/4875; automated reagent dispensing G01N35/1002
• 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 2035/0425 : Stacks, magazines or elevators for plates
• CPC Class Codes: G01N 2035/0436 : with pre-packaged reagents, i.e. test-packs
• 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. A preparatory apparatus configured to extract nucleic acid simultaneously from a number of nucleic-acid containing samples comprising:

one or more racks, each rack comprising a plurality of lanes, wherein each lane comprises a first location adjacent to and spatially separate from a second location, the first location comprising a slot shaped and sized to receive a holder, the second location comprising an aperture shaped and sized to receive a sample tube;

a plurality of sample tubes and a corresponding number of holders removably mounted in one of the one or more racks, wherein each holder is removably mounted in the rack adjacent to and spatially separate from its corresponding sample tube, wherein each sample tube is removably mounted in the rack, and wherein each sample tube is configured to accept one of the number of nucleic-acid containing samples, and wherein each holder comprises a process chamber, a waste chamber, one or more pipette tips, and one or more receptacles, wherein the one or more receptacles contain sufficient quantities of one or more reagents for carrying out extraction of nucleic acid from one of the number of nucleic-acid containing sample;

a magnetic separator positioned to apply a magnetic force to the process chamber of each holder when each holder is mounted in the rack;

a heater assembly configured to independently heat each of the process chamber of each holder when each holder is mounted in the rack; and

a liquid dispenser configured to carry out fluid transfer operations between the plurality of sample tubes and the corresponding number of holders.

2. The apparatus of claim 1, further comprising:

a sample identification verifier configured to check an identity of each of the number of nucleic-acid containing samples.

3. The apparatus of claim 2, wherein the sample identification verifier is selected from the group consisting of:

an optical character reader, a bar code reader, and a radio frequency tag reader.

4. The apparatus of claim 1, wherein the liquid dispenser has 4 heads, wherein each head is configured to accept a pipette tip from the one or more pipette tips in a holder.

5. The apparatus of claim 4, wherein no two heads accept pipette tips from the same holder.

6. The apparatus of claim 1, wherein the liquid dispenser is attached to a gantry having 3 degrees of translational freedom.

7. The apparatus of claim 1, further comprising:

electronic circuitry configured to control operation of;

the liquid dispenser;

the heater assembly; and

the magnetic separator.

8. The apparatus of claim 7, wherein the electronic circuitry controls the motion of the liquid dispenser.

9. The apparatus of claim 7, wherein the electronic circuitry is configured to cause the heater assembly to apply heat independently to the process chambers.

10. The apparatus of claim 7, wherein the electronic circuitry is configured to cause the magnetic separator to move repetitively relative to the process chamber of each holder.

11. The apparatus of claim 1, wherein the holders are suitable for a single use and are disposable.

12. The apparatus of claim 1, wherein the magnetic separator is configured to move repetitively relative to the process chamber of each holder.

13. The apparatus of claim 1, having two racks.

14. The apparatus of claim 1, wherein the number of nucleic-acid containing samples is 12.

15. The apparatus of claim 1, further comprising a second module configured to receive and to store nucleic acid extracted from the number of nucleic-acid containing samples, wherein the second module has separate locations for nucleic acid extracted from each of the nucleic-acid containing samples.

16. The apparatus of claim 1, further comprising a second module configured to receive and to store nucleic acid extracted from the number of nucleic-acid containing samples, wherein the second module comprises a separate microfluidic PCR cartridge for nucleic acid extracted from each of the number of nucleic-acid containing samples.

17. The apparatus of claim 16, wherein the microfluidic PCR cartridge contains lyophilized PCR reagents.

18. The apparatus of claim 16, wherein the microfluidic PCR cartridge contains a first lyophilized PCR reagents suitable for detecting a first analyte, and a second lyophilized PCR reagents suitable for detecting a second analyte.

19. The apparatus of claim 1, further comprising a second module configured to receive and to store nucleic acid extracted from the number of nucleic-acid containing samples, wherein the second module comprises a Peltier cooler.

20. The apparatus of claim 1, further comprising a second module configured to receive and to store nucleic acid extracted from the number of nucleic-acid containing samples, wherein the second module has a plurality of areas for receiving nucleic acid extracted from the number of nucleic-acid containing samples.

21. The apparatus of claim 20, wherein the plurality of areas are configured to be cooled independently of one another.

22. The apparatus of claim 20, wherein the apparatus comprises more than one rack.

23. The apparatus of claim 1, wherein each of the holders comprises a pipette tip sheath configured to surround the one or more pipette tips.

24. The apparatus of claim 23, wherein the pipette sheath is configured to hold 4 pipette tips.

25. The apparatus of claim 1, wherein each of the holders comprises an identifier.

26. The apparatus of claim 25, wherein the identifier is a bar-code.

27. The apparatus of claim 1, wherein each of the holders contains lyophilized sample preparation reagents.

28. The apparatus of claim 1, wherein one of the one or more reagents comprises:

a wash buffer, a release buffer, or a neutralization buffer.

29. The apparatus of claim 1, wherein the process chamber of each holder has a stellated ridge-pattern on its interior bottom surface.

30. The apparatus of claim 1, wherein the apparatus is configured to carry out extraction of nucleic acid from the number of nucleic-acid containing samples in less than about 30 minutes.

31. The apparatus of claim 1, wherein the one or more racks are removable.

32. The apparatus of claim 1, wherein each rack is configured to accept the number of nucleic-acid containing samples and the corresponding number of holders when disposed outside of the apparatus.

33. The preparatory apparatus of claim 1, wherein the apparatus is portable or can be situated on a laboratory bench-top.

34. The preparatory apparatus of claim 1, wherein the number of nucleic-acid containing samples are each independently selected from the group consisting of:

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

35. The preparatory apparatus of claim 1, wherein the liquid dispenser is configured to carry out fluid transfer operation on two or more lanes simultaneously, said fluid transfer operations requiring only two-dimensional movement of the liquid dispenser in each lane.