Method for operating a system for the integrated and automated analysis of DNA or protein

• US 10,184,946 B2
• Filed: 08/18/2015
• Issued: 01/22/2019
• 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: Gumbrecht, Walter, Paulicka, Peter
• Primary Examiner(s): Dauner, Joseph G.
• Application Number:
  US14/828,866
• Publication Number:
  US 20160041193A1
• Time in Patent Office: 1,253 Days
• Field of Search: None
• 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

A method for the integrated and automated analysis of DNA or protein, including a single-use cartridge, with an analysis device having a control device, and devices for capturing and processing signals, the control device carrying out a completely automatic process and evaluation of molecular diagnostic analysis via single-use cartridges (Lab-on-a-Chip). Controlling of an analysis process, which occurs in the cartridge, involve the subsequent displacement and thermostatisation of liquids with a first device, and with a second device the signals which are obtained during the analysis are processed. The first and the second devices are synchronized 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 method for operating a control device in a biochip analysis device comprising:

a cartridge in the form of a plastic card which contains a system of microfluidic channels that form a reservoir that contains a reaction substance, a sample chamber and flow channels, a fluid input and a deformable means for covering the system of microfluidic channels,

a control device having a cartridge receptacle for receiving, locking, unlocking and releasing the cartridge, the control device comprising;

a power supply connected to the cartridge receptacle,

at least one first sensor adapted for identifying whether or not a cartridge located in the cartridge receptacle is a correct cartridge or correctly located therein,

a microcontroller adapted for control and data acquisition, and having means for transmitting data to an evaluation unit or computer, and

a pump controlled by the microcontroller and adapted to direct a flow of liquid through a cartridge located in the cartridge receptacle,

wherein the cartridge receptacle is comprised of upper and lower parts that are movable apart for insertion of a cartridge and together for locking an inserted cartridge in the cartridge receptacle, the method comprising the steps of;

introducing a sample into said cartridge;

opening the cartridge receptacle;

using a means for moving to move a part of the cartridge receptacle upon insertion of the cartridge into the cartridge receptacle, such that the cartridge is correctly received in the cartridge receptacle;

transferring the sample to a PCR chamber;

dissolving dry PCR reagents;

closing off valves of the PCR chamber;

starting thermocycling;

carrying out a set number of thermocycles at defined temperatures;

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

transferring the PCR product from the PCR chamber to a detection chamber;

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

the carrying out the DNA analysis comprising actively changing the temperature in the detection chamber, such that hybridization of the PCR product with catcher molecules can take place;

pumping dissolved enzyme into the detection chamber;

pumping dissolved substrate into the detection chamber;

starting electrical detection; and

evaluating measurement signals resulting from the analysis or transmitting data to an external device or computer.

2. The method as claimed in claim 1, wherein an SNP analysis is carried out by recording melting curves.

3. The method as claimed in claim 1, wherein the control device comprises said means for moving a part of the cartridge receptacle and further comprising a means for locking a cartridge inserted into the cartridge receptacle.

4. Method as claimed in claim 1, wherein the control device comprises fill level sensors adapted for monitoring of the flow of fluid in the cartridge.

5. Method as claimed in claim 1, wherein a thermostatic regulation device is provided that has thermal coupling plates at positions adapted for transferring heat to a cartridge in the cartridge receptacle, one of the thermal coupling plates on a side facing the film of an inserted cartridge.

6. Method as claimed in claim 1, further comprising bringing two parts of the cartridge receptacle together by said means for moving.

7. Method as claimed in claim 1, further comprising bringing a Peltier element into position by said means for moving.

8. Method as claimed in claim 1, further comprising performing redox cycling in the detection chamber for detection of DNA.

9. Method as claimed in claim 1, wherein the closing of the valves of the PCR chamber is performed by actuators pressing onto the deformable means for covering the system of microfluidic channels of the cartridge.

10. A method for controlling an integrated and automated analysis of DNA or protein in a system comprising a cartridge containing various reagents for the analysis and to receive a sample to be analyzed and comprising a control device for controlling an analysis process, the method comprising:

introducing a sample into the cartridge;

determining, using a capacitive filling level sensor, a liquid level in a channel of the cartridge;

actively influencing the temperature in a detection chamber of the cartridge by a Peltier element of the control device which is thermally coupled to a chip module;

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

recording measurement data from the analysis process via a computing unit;

transmitting the measurement data to an evaluation unit via a wireless module; and

evaluating the measurement data; and

representing the results of the analysis on a display,

further comprising using a means for moving for bringing the Peltier element into position for said influencing of the temperature in the detection chamber.

11. Method according to claim 10, further comprising:

controlling all components of the control device involved in the analysis process via a microcontroller.

12. Method according to claim 11, wherein the microcontroller is connected to the computing unit via a serial interface.

13. Method according to claim 10, further comprising:

controlling a pump for pumping a fluid through channels of the cartridge.

14. Method according to claim 10, further comprising:

controlling at least one valve arranged in a channel of the cartridge.

15. Method according to claim 10, further comprising:

controlling valves of a PCR chamber via a servomotor and a signal processing unit.

16. Method according to claim 10, further comprising:

monitoring the movement of a fluid in the cartridge.

17. Method according to claim 10, wherein carrying out the analysis includes performing a DNA analysis, and the DNA analysis includes cell disruption and PCR.

18. Method as claimed in claim 17, wherein an SNP analysis is carried out by recording melting curves.

19. Method according to claim 10, further comprising:

carrying out a thermocycling process for PCR, wherein dry PCR reagents are dissolved.

20. Method as claimed in claim 10, wherein a protein analysis is carried out.

21. Method as claimed in claim 10, further comprising closing valves of a PCR chamber by actuators pressing onto a deformable means for covering the cartridge.

22. Method as claimed in claim 10, further comprising actively influencing the temperature in the detection chamber by a Peltier element which is thermally coupled to a chip module in order to support a hybridization between the replicated DNA and corresponding capture molecules that takes place at a surface of a chip module of the cartridge.

23. A method for controlling an integrated and automated analysis of DNA or protein in a system comprising a cartridge containing various reagents for the analysis and to receive a sample to be analyzed and comprising a control device for controlling an analysis process, the method comprising:

introducing a sample into the cartridge;

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

closing off a PCR chamber of the cartridge by closing two valves by actuators pressing onto a film of the cartridge;

pressing a device for transferring heat to the PCR chamber against a plastic body of the cartridge when a PCR is carried out, wherein said device comprises two Peltier units which are arranged on opposite sides of the PCR chamber;

carrying out a thermocycling process for PCR, wherein dry PCR reagents which are stored in the cartridge are dissolved;

carrying out a predetermined number of thermocycles at defined temperatures;

recording measurement data from the analysis process via a computing unit;

transmitting the measurement data to an evaluation unit via a wireless module; and

evaluating the measurement data,

further comprising using a means for moving to bring the device for transferring heat into position against the plastic body.

24. Method as claimed in claim 23, further comprising inserting the cartridge into a cartridge receptacle and bringing two parts of the cartridge receptacle together by said means for moving.

25. Method as claimed in claim 23, further comprising actively influencing the temperature in a detection chamber of the cartridge by a Peltier element which is thermally coupled to a chip module in order to support a hybridization between replicated DNA and corresponding capture molecules that takes place at a surface of a chip module of the cartridge.

26. Method as claimed in claim 25, further comprising performing redox cycling in the detection chamber for detection of DNA.

27. Method as claimed in claim 23, further comprising electrically detecting the PCR product by means of a sensor array.

28. A method for controlling an integrated and automated analysis of DNA in a system comprising a cartridge containing various reagents for analysis and receiving a sample to be analyzed and comprising a control device for controlling an analysis process and having Peltier elements, the method comprising:

introducing a sample to be analyzed into the cartridge;

opening a cartridge receptacle of the control device;

using a motor means for moving a part of a cartridge receptacle upon insertion of the cartridge and/or bringing two parts of the cartridge receptacle together when the cartridge has been completely introduced;

upon insertion of the cartridge, bringing Peltier elements of the control device into position by another motor means;

transferring a sample to be analyzed to a PCR chamber of the cartridge;

closing off the PCR chamber using actuators by pressing two valves onto a film of the cartridge;

performing PCR by carrying out a predetermined number of thermocycles;

opening the PCR chamber after conclusion of the PCR;

transferring the PCR product from the PCR chamber to a detection chamber of the cartridge;

actively influencing the temperature in the detection chamber using a Peltier element which is thermally coupled to a chip module in order to support hybridization between the PCR product and corresponding capture molecules that takes place at a surface of the chip module of the cartridge;

performing redox cycling in the detection chamber for detection of DNA;

electrically detecting the PCR product by means of a sensor array;

recording measurement data from the analysis process with a computing unit;

evaluating the measurement data; and

representing the results of the analysis on a display.

29. Method as claimed in claim 28, further comprising transmitting the measurement data to an evaluation unit via a wireless module.

30. Method as claimed in claim 28, further comprising interrupting the movement of the cartridge by a limit switch when the cartridge has been completely introduced.

31. Method as claimed in claim 28, further comprising marking DNA linked to catcher molecules by a label enzyme.

32. Method as claimed in claim 28, further comprising pressing Peltier units, which are arranged on opposite sides of the PCR chamber for transferring heat to the PCR chamber, against a plastic body of the cartridge when PCR is carried out.

33. Method as claimed in claim 28, wherein a Peltier element for actively influencing the temperature in the detection chamber is pressed against the cartridge.

34. Method as claimed in claim 28, wherein the sensor array comprises a CMOS sensor chip.