Microchip matrix device for duplicating and characterizing nucleic acids

• US 20020150933A1
• Filed: 01/02/2002
• Published: 10/17/2002
• Est. Priority Date: 07/02/1999
• Status: Active Grant

• Current Assignee: Alere Technologies GmbH (Abbott Laboratories Incorporated)
• Original Assignee: Alere Technologies GmbH (Abbott Laboratories Incorporated)
• Inventors: Ermantraut, Eugen, Schulz, Torsten, Ehricht, Ralf, Ellinger, Thomas, Tuchscherer, Jens, Poser, Siegfried
• Granted Patent: US 7,888,074 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01F 33/3031 using electro-hydrodynamic [EHD] or electro-kinetic [EKI] phenomena to mix or move the fluids
• CPC Class Codes: B01L 2200/142 Preventing evaporation
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0809 rectangular shaped
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0418 electro-osmotic flow [EOF]
• 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 3/508 rigid containers not provided for above

Abstract

The aim of the invention is to provide a device for duplicating and characterizing nucleic acids almost simultaneously and with a high sample throughput rate. The device consists of a chamber body with a recess whose edge sealingly holds an optically transparent chip. Said chip holds nucleic acids in individual spots on a detection surface. The chamber body is placed on an optically transparent chamber support with a bearing surface, in such a way that a capillary gap, which can be filled with a liquid sample, is formed between the detection surface of the chip facing towards the chamber support and said chamber support. The chamber body is provided with an inlet and an outlet, which are spatially separate from each other, and has a space, which laterally encompasses the chip and which has a gas reservoir. The chamber support is provided with heating elements.

Claims

1. Device for duplicating and characterizing nucleic acids in a reaction chamber,

characterized in that a chamber body (1) containing an optically permeable chip (2) having a detection area (12), and being optically permeable at least in the zone of the detection area (12) of the chip (2), is sealingly placed on an optically permeable chamber support (5), so that a sample chamber (3) having a capillary gap (7) is formed between the chamber support (5) and the detection area (12) of the chip (2), which is temperature-adjustable and flow-controllable.

2. Device according to claim 1,

characterized in that the temperature adjustment means are connected with the chamber support (5) and permit a rapid heating and/or cooling of the sample chamber (3) having the capillary gap (7).

3. Device according to claim 2,

characterized in that the temperature adjustment means are situated on the side of the chamber support (5) facing towards the chamber body (1).

4. Device according to any one of the preceding claims,

characterized in that the temperature adjustment means (16, 17) are configured in the form of optically transparent thin films and/or are so finely structured that the optical transparency of the chip (2) remains unaffected at least in the zones of the spots (13) of the detection area (12).

5. Device according to claim 4,

characterized in that the temperature adjustment means comprise micro-structured heating elements (17), preferably nickel-chromium thick film resistance heaters and/or micro-structured temperature sensors (16), preferably nickel-chromium thick film resistance sensors.

6. Device according to any one of the preceding claims,

characterized in that the chamber support (5) comprises systems for thoroughly mixing the liquid sample, which are configured in the form of optically transparent thin films and/or are so finely structured that the optical transparency of the chip (2) remains unaffected at least in the zones of the spots (13) of the detection area (12), whereby preferably a quadrupole system for inducing an electro-osmotic flow is concerned.

7. Device according to claim 6,

characterized in that the quadrupole system is realized as gold-titanium electrodes.

8. Device according to any one of the preceding claims,

characterized in that the chamber support (5) and the chamber body (1) preferably consist of glass and/or synthetic material and/or optically permeable synthetic materials particularly preferred of nylon, Teflon, topaz, polycarbonate, polystyrene, PMMA and/or polymethane ethyl acrylate.

9. Device according to any one of the preceding claims,

characterized in that the chamber support (5) consists of a thermally conducting material.

10. Device according to any one of the preceding claims,

characterized in that the chip consists of optically permeable materials, preferably of glass, borofloat glass, quartz glass, monocrystalline CaF₂, sapphire, PMMA and/or silicon.

11. Device according to any one of the preceding claims,

characterized in that the chamber body (1) comprises, at least in the zone of the chip (2) an optically permeable conical recess.

12. Device according to any one of the preceding claims,

characterized in that the chamber body disposes of an inlet (81) and an outlet (82) spatially separate from each other, for charging the sample chamber (3) and the capillary gap (7).

13. Device according to claim 12,

characterized in that the inlet (81) and the outlet (82) are arranged unilaterally to the chip (2) and are separated by a gas reservoir nose (9).

14. Device according to any one of the preceding claims,

characterized in that the chamber body (1) is sealingly and unreleasably connected with the chamber support (5) by an adhesive and/or weld connection, or is releasably connected through an additional sealing surface (43).

15. Device according to any one of the preceding claims,

characterized in that the detection area (12) is configured in the form of spots, onto which probes (56, 57, 58, 59) in the form of nucleic acid molecules are immobilized, said nucleic acid molecules preferably being DNA molecules and/or RNA molecules.

16. Device according to claim 15,

characterized in that the probes (56, 57, 58, 59) are immobilized through spacers (55).

17. Device according to any one of claims 1 through 14,

characterized in that the detection area (12) is configured in the form of spots, onto which probes (56, 57, 58, 59) in the form of peptides and/or proteins are immobilized, preferably antibodies, receptor molecules, hormones and/or pharmaceutically active peptides being concerned.

18. Device according to any one of the preceding claims,

characterized in that the evaluation of the chip-based characterization may ensue by forms of the optical detection and/or spectroscopy, particularly preferred by transmitted-light fluorescence measurement, dark field fluorescence measurement, confocal fluorescence measurement, reflected-light fluorescence measurement, photometry and/or differential photometry.

19. Device according to any one of claims 1 through 17,

characterized in that the evaluation of the chip-based characterization ensues by a silver precipitation reaction.

20. Use of a device according to any one of the preceding claims for an almost simultaneous performance of reprocessing reactions and/or conditioning reactions and a chip-based characterization of the products.

21. Use of a device according to claim 20, the reprocessing reaction and/or conditioning reaction concerning an amplification of nucleic acids by PCR.

22. Use of a device according to claim 20, the reprocessing reaction and/or conditioning reaction concerning a reverse transcription of RNA to cDNA.

23. Use of a device according to any one of claims 1 through 17, the reprocessing reaction and/or conditioning reaction concerning a digestive process of nucleic acids by means of restriction enzymes.

24. Use of a device according to claim 20 for the almost simultaneous amplification of DNA by PCR and for the chip-based characterization of the PCR products.