System for the Integrated and Automated Analysis of DNA or Protein and Method for Operating Said Type of System

• US 20090298059A1
• Filed: 05/22/2006
• Published: 12/03/2009
• Est. Priority Date: 05/25/2005
• Status: Active Grant

• Current Assignee: Boehringer Ingelheim Vetmedica GmbH (C.H. Boehringer Sohn AG & Co. KG)
• Original Assignee: Boehringer Ingelheim Vetmedica GmbH (C.H. Boehringer Sohn AG & Co. KG)
• Inventors: Paulicka, Peter, Gumbrecht, Walter
• Granted Patent: US 9,110,044 B2
• Time in Patent Office: Days
• US Class Current: 435/6.11
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• 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 2200/143 Quality control, feedback systems
• CPC Class Codes: B01L 2300/023 Sending and receiving of information, e.g. using bluetooth
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1877 using chemical reactions
• CPC Class Codes: B01L 2400/0406 capillary forces
• 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 3/502723 characterised by venting arrangements
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• 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 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 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: G01N 2035/00019 cassette structures
• CPC Class Codes: G01N 2035/00346 Heating or cooling arrangements
• CPC Class Codes: G01N 2035/00376 Conductive heating, e.g. heated plates
• CPC Class Codes: G01N 2035/00841 : results of the analyses
• CPC Class Codes: G01N 2035/1034 : Transferring microquantities of liquid
• CPC Class Codes: G01N 35/00009 : provided with a sample supporting tape, e.g. with absorbent zones
• CPC Class Codes: G01N 35/00584 : Control arrangements for automatic analysers
• CPC Class Codes: G01N 35/00732 : Identification of carriers, materials or components in automatic analysers
• CPC Class Codes: G01N 35/00871 : Communications between instruments or with remote terminals
• CPC Class Codes: G01N 35/08 : using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
• CPC Class Codes: G01N 35/1002 : Reagent dispensers

Abstract

An embodiment of the present invention relates to a system for the integrated and automated analysis of DNA or protein, including a single-use cartridge, an analysis device comprising a control device, and devices for capturing and processing signals. An embodiment of the present invention relates, in particular, to the control device for carrying out a completely automatic process and evaluation of molecular diagnostic analysis via single-use cartridges (Lab-on-a-Chip). The first devices are provided for controlling an analysis process which occurs in the cartridge, subsequently the displacement and the thermostatisation of liquids, and the second devices are provided for processing the signals which are obtained during the analysis. The first and the second devices are synchronised in such a manner that the analysis process of the sample can be carried out in a totally integrated manner thus producing an immediate result.

Claims

1. A system for the integrated and automated analysis of DNA or protein, comprising:

a cartridge to receive a sample and various reagents suitable for the analysis in dry and long-term-stable form;

an evaluation device, containing a control device to perform an analysis process when the cartridge is introduced into the evaluation device;

means for processing the sample within the cartridge, for which purpose liquid from a liquid container is fed at least to the dry reagents in the cartridge for the purpose of dissolving and subsequently detecting sample constituents to be detected;

first means for controlling introduction of the cartridge into the analysis device and the subsequent analysis process proceeding in the cartridge including movement and thermostatic regulation of liquids introduced into the cartridge; and

second means for processing signals obtained through the analysis process are present,

the first and second means, in the control device, being coordinated with one another in such a way that the analysis process of the sample can be carried out in a fully integrated manner and the signals are directly outputtable.

2. The system as claimed in claim 1, wherein interfaces for the fluidics, on the one hand, and the electronics, on the other hand, are present between the cartridge and the control device.

3. The system as claimed in claim 2, wherein the control device comprises at least one of at least one serial and one wireless interface via which at least one of the signals and at least partly processed signals are communicatable.

4. The system as claimed in claim 3, wherein the wireless interface is formed as a Bluetooth system.

5. The system as claimed in claim 1, further comprising an evaluation server.

6. The system as claimed in claim 1, further comprising a PDA.

7. The system as claimed in claim 1, further comprising a cartridge receptacle to receive, lock, unlock and release the cartridge.

8. The system as claimed in claim 7, further comprising sensors to identify a correct orientation of the cartridge in the cartridge receptacle.

9. The system as claimed in claim 8, further comprising sensors to identify a correct type of the cartridge in the cartridge receptacle.

10. The system as claimed in claim 1, further comprising a water reservoir to dilute the sample, to dissolve dry reagents stored in the cartridge and to carry out flushing operations in the control device.

11. The system as claimed in claim 10, wherein fluidic contact is made with the water reservoir by way of a septum being pierced by a cannula.

12. The system as claimed in claim 10, further comprising a filter to ventilate the water reservoir when water is drawn.

13. The system as claimed in claim 1, further comprising:

a power supply unit.

14. The system as claimed in one claim 1, further comprising:

a pump.

15. The system as claimed in claim 1, wherein the control device comprises a line system with valves by which water is introduceable into the cartridge.

16. The system as claimed in claim 15, further comprising a pressure sensor in the line system.

17. The system as claimed in claim 15, further comprising at least one interface to feed water into the cartridge.

18. The system as claimed in claim 17, wherein the at least one interface is formed as a hollow needle.

19. The system as claimed in claim 1, further comprising means for identifying the filling of channels on the cartridge.

20. The system as claimed in claim 19, wherein the means for identifying the filling are embodied as capacitive sensors with electrodes, wherein the electrodes are arranged in such a way that, when the cartridge is introduced into the control device, one of the electrodes is situated above the cartridge and one of the electrodes is situated below the cartridge.

21. The system as claimed in claim 1, further comprising valves to close off and open venting ports of the cartridge.

22. The system as claimed in claim 1, further comprising means for closing off and opening chambers on the cartridge.

23. The system as claimed in claim 22, wherein the means for closing off and opening the chambers are embodied as movable plungers.

24. The system as claimed in claim 1, further comprising a combined unit composed of a magnetic bead collector for capturing magnetic beads and a thermostatic regulation device for carrying out thermocycling processes for carrying out a PCR in the control device.

25. The system as claimed in claim 24, wherein the thermostatic regulation device comprises two thermostatic regulation modules arranged in a sandwich arrangement, and wherein the cartridge in the inserted state is situated between the thermostatic regulation modules.

26. The system as claimed in claim 25, further comprising means for generating a defined contact pressure between the thermostatic regulation modules and the cartridge.

27. The system as claimed in claim 1, further comprising means for making electrical contact with an electrical chip module.

28. The system as claimed in claim 27, further comprising a thermostatic regulation unit for the chip module.

29. The system as claimed in claim 1, further comprising control, measuring and regulating electronic devices for the functions.

30. The system as claimed in claim 1, further comprising a microcontroller for control and data acquisition.

31. The system as claimed in claim 1, further comprising a display for representing the analysis results.

32. A method for operating the control device in the system claimed in claim 1, comprising:

introducing a sample into a cartridge;

carrying out an analysis of constituents to be detected in the sample in an integrated process; and

evaluating measurement signals resulting from the analysis operation.

33. The method as claimed in claim 32, wherein a cell disruption and a PCR are carried out for a DNA analysis.

34. The method as claimed in claim 33, further comprising:

opening the cartridge receptacle and using the pump to draw water from the reservoir given a suitable valve position;

inserting, via a user, the cartridge with a sample;

automatically starting the analysis;

pumping water into the cartridge via a water port to wash out excess sample material;

diluting, as a result of the inflow of water, the sample and pumping it into the cell disruption channel, wherein dry reagents are dissolved, wherein;

a residence time ensues for cell disruption, and wherein liberated DNA binds to magnetic beads;

filing, nearly at the same time two ELISA reagent channels, wherein dry ELISA reagents are dissolved;

flushing the content of the cell disruption channel by further pumping of water through the PCR chamber, wherein the magnetic beads with DNA are held back by a magnetic field, the remainder is flushed into a waste channel;

closing off the valves of the PCR chamber are closed off;

starting the thermocycling is sated, wherein dry PCR reagents are dissolved;

carrying out a predetermined number of thermocycles at defined temperatures;

opening the valves of the PCR chamber after conclusion of the PCR;

pumping water into the cartridge via a further water port and flushing the PCR product from the PCR chamber into a detection chamber;

thermostatically regulating the detection chamber, wherein hybridization of the DNA with catcher molecules takes place;

pumping air possibly present between the PCR product and the dissolved ELISA reagents into a waste channel;

pumping the dissolved enzyme label via the detection chamber into a further waste channel;

pumping the dissolved substrate via the detection chamber into a further waste channel; and

starting the electrical detection.

35. The method as claimed in claim 33, wherein an SNP analysis is carried out by recording melting curves.

36. The method as claimed in claim 32, wherein a protein analysis is carried out.

37. The method as claimed in claim 32, wherein a hapten analysis is carried out.

38. The system as claimed in claim 11, further comprising a filter to ventilate the water reservoir when water is drawn.

39. The system as claimed in claim 16, further comprising at least one interface to feed water into the cartridge.

40. The method as claimed in claim 34, wherein an SNP analysis is carried out by recording melting curves.