Molecular diagnostics system and methods

• US 8,883,487 B2
• Filed: 12/21/2005
• Issued: 11/11/2014
• Est. Priority Date: 12/23/2004
• Status: Expired due to Fees

• Current Assignee: Abbott Point Of Care Incorporated (Abbott Laboratories)
• Original Assignee: Abbott Point Of Care Incorporated (Abbott Laboratories)
• Inventors: Collier, Gordon Bruce, MacLeod, Jason Andrew, Nemeth, Attila Csaba, Miller, Cary James, Wood, John Allister, Dicke, William Charles
• Primary Examiner(s): HOBBS, MICHAEL L
• Application Number:
  US12/308,307
• Publication Number:
  US 20100291666A1
• Time in Patent Office: 3,247 Days
• Field of Search: 435/287.2, 435/6.1
• US Class Current: 435/287.2
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• 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 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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 3/502769 characterised by multiphase flow arrangements
• 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: B03C 1/01 by addition of magnetic adjuvants
• CPC Class Codes: B03C 1/0332 using permanent magnets
• CPC Class Codes: B03C 1/0335 using coils
• CPC Class Codes: B03C 1/288 disposed at the outer circumference of a recipient
• CPC Class Codes: B03C 1/30 Combinations with other devices, not otherwise provided for
• CPC Class Codes: B03C 2201/18 Magnetic separation whereby the particles are suspended in a liquid
• CPC Class Codes: B03C 2201/26 for use in medical or biological applications
• CPC Class Codes: C12N 15/1003 Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
• CPC Class Codes: C12N 15/1006 by means of a solid support carrier, e.g. particles, polymers
• CPC Class Codes: C12N 15/1013 by using magnetic beads
• CPC Class Codes: C12Q 1/6844 : Nucleic acid amplification reactions
• CPC Class Codes: C12Q 1/6853 : using modified primers or templates
• CPC Class Codes: C12Q 1/686 : Polymerase chain reaction [PCR]

Abstract

The present invention relates to automated devices and methods for the extraction of nucleic acids from cells, the amplification of segments of nucleic acid and the detection of nucleic acids, all in a convenient and portable manner. The invention is particularly suited for use in point-of-care medical diagnostic testing.

Claims

1. An integrated single-use device for performing a nucleic acid analysis, comprising:

a housing,

an entry port for accepting a sample suspected of containing a target nucleic acid,

a first chamber operably connected to the entry port and comprising;

a reagent for extracting the target nucleic acid,

magnet beads comprising a coating configured to bind the target nucleic acid, and

a wax configured to melt and form a contiguous wax layer in a region of egress of the first chamber;

a means for generating a magnetic field configured to draw the magnet beads through the contiguous wax layer;

a first conduit permitting passage of extracted nucleic acid from the region of egress of the first chamber into an amplification chamber, said housing containing an amplification reagent that is capable of incorporating a detectable label into an amplified nucleic acid target,

the amplification chamber having a heating means and a temperature sensing means for controlling amplification conditions,

the amplification chamber being operably linked to a second conduit containing a sensing region with an immobilized capture oligonucleotide,

a means for moving the amplified target to the sensing region to permit binding of said amplified target to said capture oligonucleotide, and

a holding chamber operably attached to said second conduit and containing a fluid configured to substantially displace uncaptured amplified target from the sensing region and permit sensing of said detectable label retained in said sensing region.

2. The device of claim 1, wherein the label moiety is selected from the group consisting of fluorocein, dinitrophenol, biotin, cholesterol, digoxigenin and estrialdiol.

3. The device of claim 1, wherein the sensing region is interrogated by an optical detector.

4. The device of claim 1, wherein the sensing region is interrogated by an integrated optical detector.

5. The device of claim 1, wherein the sensing region is interrogated by an integrated electrochemical detector.

6. The device of claim 1, wherein amplification reagent is located in said first conduit.

7. The device of claim 1, wherein amplification reagent is located in said amplification chamber.

8. The device of claim 1, wherein amplification reagent is located in a holding chamber operably connected to said amplification chamber.

9. The device of claim 1, wherein amplification reagent is located in a holding chamber operably connected to said first conduit.

10. The device of claim 1, wherein said fluid is a wash fluid and contained in a sealed pouch within the holding chamber.

11. The device of claim 1, wherein said means for moving the amplification target is pneumatic.

12. An integrated single-use device for performing a nucleic acid analysis, comprising:

a housing,

an entry port for accepting a sample suspected of containing a target nucleic acid,

a first chamber operably connected to the entry port and comprising;

a reagent for extracting the target nucleic acid onto magnetic beads that are located in the first chamber, and

a wax configured to melt and form a substantially contiguous wax layer that covers an egress region of the first chamber into a first conduit,

wherein the magnetic beads that are associated with the extracted target nucleic acid are configured to pass through the wax layer into the first conduit by means of an applied magnetic field,

the first conduit permitting passage of the extracted target nucleic acid and the magnetic beads into an amplification chamber, said housing containing an amplification reagent that is capable of incorporating a detectable label into an amplified nucleic acid target,

the amplification chamber having a heating means and a temperature sensing means for controlling amplification conditions,

the amplification chamber being operably linked to a second conduit containing a sensing region with an immobilized capture oligonucleotide,

said housing containing a means for moving the amplified target to the sensing region to permit binding of said amplified target to said capture oligonucleotide, and

a holding chamber operably attached to said second conduit and containing a fluid configured to substantially displace uncaptured amplified target from the sensing region and permit optical sensing of the detectable label retained in the sensing region.

13. The device of claim 12, wherein the detectable label is selected from the group consisting of fluorocein, dinitrophenol, biotin, cholesterol, digoxigenin and estrialdiol.

14. The device of claim 12, wherein the sensing region is interrogated by an optical detector.

15. The device of claim 12;

wherein the sensing region is interrogated by an integrated optical detector.

16. The device of claim 12, wherein amplification reagent is located in said first conduit.

17. The device of claim 12, wherein amplification reagent is located in said amplification chamber.

18. The device of claim 12, wherein amplification reagent is located in a holding chamber operably connected to said amplification chamber.

19. The device of claim 12, wherein amplification reagent is located in a holding chamber operably connected to said first conduit.

20. The device of claim 12, wherein said fluid is a wash fluid and contained in a sealed pouch within the holding chamber.

21. The device of claim 12, wherein said means for moving the amplification target is pneumatic.