EP1807208A1

EP1807208A1 - Arrangement for integrated and automated dna or protein analysis in a single-use cartridge, method for producing such a cartridge and operating method for dna or protein analysis using such a cartridge

Info

Publication number: EP1807208A1
Application number: EP05801513A
Authority: EP
Inventors: Heike Barlag; Siegfried Birkle; Walter Gumbrecht; Daniela KÜHN; Peter Paulicka; Manfred Stanzel
Original assignee: Siemens AG
Current assignee: Siemens AG
Priority date: 2004-10-15
Filing date: 2005-10-17
Publication date: 2007-07-18
Anticipated expiration: 2025-10-17
Also published as: WO2006042734A1; CN101039750A; JP2008517259A; EP1796838B1; EP1807208B1; WO2006042838A1; US20090136922A1; CN101039751B; US20090130658A1; EP1796838A1; US7851227B2; CN101039751A; JP4546534B2; CN100534619C

Abstract

A cartridge (card) having a system of microchannels and/or microcavities is used for automated DNA or protein analysis, wherein said microchannels or microcavities comprise geometrical structures for receiving dry reagents. For the purpose of industrial production, the cartridge is produced from a flat card support, e.g., by injection moulding. The reagents are spotted into the open channels, dried and then the channels are sealed by means of a film. A finished cartridge can thus be provided with a test sample and the fully automated measuring sequence can be initiated by inserting said cartridge into a read-out device.

Description

description

Arrangement for integrated and automated DNA or protein analysis in a single-use cartridge, production method for such a cartridge and method of operating the DNA or protein analysis using such a cartridge

The invention relates to an arrangement for integrated and automated DNA or protein analysis in a single-use cartridge. Cartridge is a flat card in credit card format. In addition, be ^¬ the invention contemplates the production of such a cartridge. Finally, the invention also relates to an operating method for DNA or protein analysis using such a cartridge.

For nucleic acid analysis, for example for the analysis of white blood cells from whole blood, for the purpose of answering hu mangenomischen questions the cells ^¬ must be broken first in a ers¬ th station as a sample preparation step and then the DNA iso ^¬ be profiled liberated. In a second station, a PCR (polymerase chain reaction) for selective DNA amplification (amplification) is carried out in order to increase the concentration of the DNA to be detected to such an extent that it is detected in a third station can be.

In the laboratory, the latter sub-processes are carried out separately according to the known state of the art. The three stations explained above each involve several work steps and are performed independently of each other with different devices. Carried out the individual steps weitge ^¬ starting manually.

The realization of these steps is the presence of laboratory equipment - such as a cell disruption apparatus, a PCR device (so-called thermocycler), possibly a PCR device, which is for Quantitative PCR is suitable for an electrophoresis Appara ^¬ ture, a hybridization station, an optical reader, so-called Eppendorf tubes, multiple pipetting equipment including cooling containers for reagents -. dependent and by trained personnel in compliance with safety regulations must be¬ züglich Danger of infection, waste disposal od. Like. Durchge leads. In particular, several volumetric, ie precise, dosages (pipetting) of reagent solutions must be carried out. Such steps are time consuming and costly.

Devices are known for biochemical analysis of the prior art, which accordingly make dung WO 02/073153 Al USAGE ^¬ of particularly silicon-based measurement modules, which can be integrated in a chip card. Here, whoever the WO 02/072262 Al already lysis module integrated in accordance with ^¬ the reagents used for analysis in dry-stored form into the Ana¬.

On this basis, the object of the invention is the realization of a low-cost, easy-to-handle, complete DNA or protein analysis process in a miniaturized cartridge. In particular, the following improvements compared to the laboratory method should be realized: - A complete integration of all substances (possibly except

Water) in a closed single-use Cartrid¬ ge;

a storage of the reagents in a storage-stable form at room temperature; - ridge automatic execution of all processes in the Cart ^¬;

no manual operations, except injection of the sample to be analyzed, e.g. Blood;

- No direct contact with hazardous substances (blood and reagent waste remain in the cartridge);

- Cartridge geometry allows efficient and fast thermocycling;

- All detection processes should be electrically and be read out ein¬ easy;

- The cartridge used is small and inexpensive to produce.

The object is achieved by an arrangement with egg ^¬ ner cartridge according to claim 1. The preparation of such cartridge is the subject of claim 34. Operating methods with such cartridges are the subject of independent claims 38 and 42. In this case, at ^¬ is demanding directed to the Prote in ^¬ analysis 38 on DNA analysis and claim 42nd Further developments of the arrangement, of the production method and of the mode of operation during the analysis are specified in the respective subclaims.

The invention is based, in particular, on WO 02/072262 A1 and the further prior art mentioned therein. There, an analysis device with dry-stored in fluid channels, stable at room temperature reagents is described, which are brought into solution by supplying water shortly before their intended use. The invention is furthermore based on the unpublished DE 10 2004 021780 A1 and the unpublished DE 10 2004 021822 A1. In addition, it is also known to use specially electrically readable detection modules.

In contrast, the subject of the invention is such a disposable cartridge with a system of microchannels and / or microcavities for a given process flow after sample taking, the cartridge having structures for receiving the dry reagents and these structures means for performing both the cell digestion on the one hand and the PCR on the other hand, but also the electrochemical detection are associated. In particular, the channels have different, problem-adapted structures. Spe ^¬ essential the digestion channel advantageously has stepped cross sections for optimum wetting of the dry reagent, while the PCR chamber and the Elisa reagent channels have potty-shaped wells.

It can thus be achieved that, in a process sequence, the supply and preparation of the sample, the DNA

Amplification and the actual detection of the DNA is possible.

The inventive system of the geometric structures ren in microchannels or microcavities for receiving dry reagents to provide suitable conditions for DNA analysis of a hand and the other hand, protein analysis ^¬. The following features and measures are essential:

- The changes introduced in the microchannel or the microcavity reagents are dryable substances with vernachläs ^¬ sigbarem vapor pressure. The substances which are stable at room temperature retain their properties for cell digestion and / or PCR and / or detection. Here, mixtures may be formed with excipients thin films of the materials and the mixtures may thin Pa ^¬ refine layers be watertight covered.

In the invention, the reagents and auxiliaries are already introduced into depressions of the cartridge channels 4 as dry substances in the production of the cartridge. This gives the following advantages:

- easy and precise application of the reagents in the production of the cartridge

Protection of the reagents during filling of the reagent channels, i. The reagents are not washed away by a so-called water flow, but are retained when the entire channel is filled. Only after filling the channel, the reagent spots dissolve via diffusion processes and there is a homogeneous reagent solution.

In a further particular embodiment, the recesses are located at predefined intervals along the reagent channel. The distances can be equidistant or be arranged particularly advantageous in variable spacing patterns.

The depressions can advantageously be filled with variable amounts of dry reagent. Through the combination of different dry reagent quantities and well spacing patterns, the desired concentration profiles of the final reagent solutions can be adjusted.

For certain functions, such as Cell disruption in the presence of magnetic beads and lysis reagents is a uniform distribution of the insoluble components, i. of the beads, in dry reagent required. For this purpose, the magnetic beads are dispensed as a suspension in the lysis channel. Evaporation of the solvent is observed to cause the beads to enter the

Retract edge regions of the lysis channel and a gleichmä ^¬ lar distribution is thus not given. By stepwise structuring of the lysis channel cross section, the magnetic beads are distributed over the steps and a uniform distribution is achieved.

In order to be able to carry out a cell disruption, a PCR and a so-called DNA / protein ELISA test in the same way with the cartridge according to the invention, it is advantageous that in the microchannels or microcavities substrates with DNA-binding properties, in particular the DNA-binding Magnetic beads are present. The lysis reagents and the magnetic beads may be contained together in a single dry matrix. Furthermore, the card also contains the reagents for an ELISA assay. Two reagents are described in detail for the ELISA assay required, that the first agent Re ^¬ a label enzyme, and as the second reagent is an enzyme substrate.

In particular, a detection module for the electrical detection of the hybridization processes is arranged in the cartridge. The detection module advantageously consists of an el-metal / plastic composite or a semiconductor-processed Silicon chip with precious metal electrodes. In particular, electrochemical, magnetic or piezoelectric measuring methods are suitable for electrical detection.

To apply the invention of the cartridge according to the invention is in particular an input port for a whole blood sample before ^¬ handen. Furthermore, means are for supplying water vorhan¬ to, for example, inflow ports for connection to an ex ternal ^¬ water supply or an integrated water reservoir. In the microchannels or microcavities, dry buffer substances have a defined ionic strength after the supply of water.

When applying the invention for the analysis of white blood cells from whole blood, means for mixing a whole blood sample with water or a buffer solution are advantageously present. In this case, means for flowing through the lysis / bead / reagent coated microchannel or microcavity with blood or blood / water or blood / buffer mixtures are present.

A magnetic field spe¬ for the invention in the cartridge in use essential for DNA analysis carried out PCR are still With ^¬ tel for generating for fixing the DNA / magnetic bead complex in the PCR cavity there. For this purpose, the PCR cavity must be able to be suitably closed and means for thermocyclization must be present.

Finally, in the case of the cartridge according to the invention, it is essential that there are means for storing spent sample material and spent reagents which form waste reservoirs. In this case, the means for germ-proof, cell-free or particle-free venting of at least one waste reservoir must be suitable. Finally, to read out the cartridge in a read-out device, which is not the subject of the invention, means for fixing the cartridge must be present.

Further details and advantages of the invention will become apparent from the following description of the figures Ausführungsbei¬ play with reference to the drawing in conjunction with the patent claims. They show in a schematic representation

1 shows a cartridge with an overview of individual

Microchannel / cavity systems with the associated function designations,

FIG. 2 shows the plan view of a cell disruption channel, FIG. 3 shows the cross section through the cell disruption channel, FIG. 5, FIG.

4 shows two alternatives of the flow channel cross-section in an enlarged view,

5 the top view of the PCR chamber in FIG. 1, FIGS. 6 and 7 the cross section through the PCR chamber according to FIG. 5,

Figure 8 is a plan view of an ELISA reagent channel in Fi gur ^¬ 1, 9, 10 in accordance with the cross section of the ELISA Reagenzkanals

FIG. 8 and FIGS. 11 to 23 show the plan view of the cartridge according to FIG. 1 in various process ^states during an automated evaluation.

In the figures, identical or equivalent elements have the same reference numerals. In particular, Figures 1 to 10 ei ^¬ hand, and Figures 11 to 23 on the other hand ge ^¬ be described jointly.

In Figure 1, a cartridge 100 for an ELISA ( "enzyme-linked immunosorbent assay") - test with an elevation of the existing therein microchannel or Darge cavities system ^¬ is, the clarity, the associated function ^¬ designations are also referred to , The cartridge 100 consists in detail of a plastic body 101 with there introduced fluidic structures, the one of

Plastic film are covered. The structures will be described below with reference to FIGS. 2 to 10. 1 shows a sample port 102 with adjoining metering section 105, via which defined liquid samples for, in particular, nucleic acid analysis, for example for the analysis of white blood cells from whole blood, can be obtained in order to answer human-genomic questions. This is followed by a channel region 110 for the cell disruption of the sample, and furthermore, especially for a DNA analysis, a region 120 for a PCR (Polymerase Chain Reaction) for selective DNA amplification (amplification) to increase the concentration of the DNA to be detected so far that it can be detected in a third station. The actual PCR chamber can be closed by valves 122, 122 '. The detection of the thus prepared sample, in particular according to the ELISA method, then takes place in the region 130.

From Figure 1 may also be water ports 103-103 '''he ^¬ clearly. When preparing a sample, water can be introduced into the Cartridge 100 as a transport and solvent. Furthermore, vent ports 104 to 104 '''are present.

In particular, as mentioned, the cartridge 100 has an input port 102 for a whole blood sample. For this purpose, means for supplying water are present. There may be an inflow port for connection to an external water supply or the inflow port may be connected to an integrated water reservoir.

The microchannels or microcavities 101-131 are filled in the Nor mally ^¬ with dry buffer substances which ensure a de ^¬ finierte ionic strength to the water supply. For the blood analysis are means for mixing whole blood samples and water or the buffer solution and / or means for flowing through a microsphere coated with a lysis bead reagent or the microcavity with blood or with blood-water or blood-buffer mixture available. In the channel system are wide areas 106, 107, 108, 109 is provided as re ^¬ servoirs for receiving waste ( "waste"). In addition, there is an area with channels 131 and 131 'for receiving different ELISA reagents.

In FIGS. 2 to 4, reference numerals 101 again identify the cartridge base body. The body includes a spe¬ essential for cell disruption ( "lysis") a shaped in a special way flow passage 111 with edges formed by the sides ^¬ step-shaped recesses 112 is used to receive me of dry reagents. The recesses 112 in this case have a plurality of steps with step heights of 10 to 500 .mu.m and ha ^¬ ben an extension of approximately 1 mm and a depth of about microns 100th

Especially in the illustration according to FIG. 4a, the alternative possibility arises of providing the lysis reagents only in the area 113 of the edges of the flow channel 111 in the case of a flow channel without additional depressions. On the other hand, in FIG. 4b, reagents of this kind, which in particular also contain magnetic beads for binding the released DNA, are distributed uniformly between the steps 112 via the flow channel 111. Magnetic beads have DNA- and protein-binding properties, provided that they have been pretreated accordingly. They may interfere with DNA-binding properties. optionally also be coated with antibodies. For introducing dry substances as matrix with lysis reagent and magnetic beads, reference is made in particular to the older DE 10 2004 021780 A1 and the earlier DE 10 2004 021822 of the applicant.

FIGS. 5 to 7 show the structure of a PCR chamber 120 in the cartridge main body 101 with flow channel 111. The valve arrangement for completing the PCR chamber in the case of appropriate use is not shown here. It is essential that in the PCR chamber 120 round cylindrical recesses 124, 124 'for receiving specific reagents 127, 127 ', which are needed in carrying out the PCR, are present. Specifically, it is shown in FIG. 7 that a dry-storable, room temperature-stable PCR reagent 127, 127 'is initially covered by a paraffin layer 128, 128'.

The proper implementation of PCR with valve-controlled thermocycling in a cartridge is nen in individual ^¬ in the parallel applications DE 10 2004 050576.4 and DE 10 2004 050510.1 applicant with the same application priority be ^¬ wrote, is expressly referred to in the present context ( "Incorporation by Reference "). In particular, it describes the use of magnetic beads for DNA binding and the concentration of the magnetic beads with the DNA in the PCR chamber 120 by means of controllable magnetic fields, which will not be discussed in detail here.

FIGS. 8 to 10 show the structure and the structure of the ELISA reagent channels 131 and 131 'from FIG. 1. In each case, well-shaped depressions 132 to 132 ^6' are present which are suitable for receiving pre-dosed and pre-ported quantities Reagents for the ELISA process according to FIG. 9 are suitable. This is described in detail in WO 02/072262 A1, which has already been cited at the outset as state of the art, to which reference is likewise expressly made in the present context ("Incorporation by Reference"). In Figure 10, the circular cylindrical recesses 132-132 ^{6 'with} dry reagents 133-133 ^6' filled dar¬ provided. In this case, a first reagent realizes a labeling enzyme and a second reagent an enzyme substrate, as is known to be required in the hybridization of the sample optionally prepared by a PCR with specific capture probes. In only schematically indicated area 130 of the detection may be in a module of a precious metal / plastic composite are different sensors for Erfas ^¬ solution of biochemical reactions. Especially in elektro¬ chemical measurements with semiconductor-processed chips, ie in particular silicon-based sensors, the signals electrically detected and processed immediately. In addition to electrochemical, magnetic and / or piezoelectric measuring methods with related sensors are also possible.

In FIGS. 11 to 23, the cartridge 100 according to FIG. 1 is shown in plan view, with the area of the cartridge 100 active during the analysis process being marked. For this purpose, the cartridge 100 is transformed into an analyzer, which is not shown in detail in the figures and not Gegens ^¬ tand the present patent application, is inserted.

The evaluation is illustrated below with reference to eleven specific procedural ^¬ RENS steps a) to m) after the cartridge is inserted into an evaluation unit with means for receiving the Cart ^¬ ridge and the evaluation device is activated at a fixed therein cartridge. With reference to FIG. 1, the following sub-steps result in detail:

a) About 10 μl of blood is added as a test sample. 1 μl is dosed automatically via the dosing capillary 105. b) The excess blood is washed into the void 106 (Waste 1). c) Subsequently, 1 μl of blood sample is diluted with water and transferred to the cell digestion channel 110. There, cell lysis of the blood cells and binding of the liberated DNA to the magnetic beads take place. d) Subsequently, the magnetic beads are transferred to the PCR chamber and collected there. There is a washing ^¬ process takes place, the washing solution in the void 107th

(Waste 2) is collected. e) Now the washing process is completed. f) Subsequently, the PCR chamber valves 122, 122 'are closed and the PCR is carried out. g) Similarly, the ELISA reagent channel 131, which contains the enzyme substrate, is filled with water during the PCR. h) Similarly, during the PCR of the ELISA reagent channel 131 ', which contains the label enzyme, ge filled with water ^¬ . i) After the PCR, the PCR chamber valves 122, 122 'are opened and the PCR product is passed via the detection module 130 (into the waste 3, channel 108), where the hybridization with the specific capture probes er¬ follows , j) The enzyme-substrate channel is vented into the waste channel 108 (waste 3). k) The label enzyme channel is in the waste channel 108th

(Waste 3) vented. 1) The label enzyme solution flows over the detection module

130 to Labein in the waste area 109 (Waste 4). m) The enzyme-substrate solution flows via the Detektionsmo ^¬ module 130 for enzymatic-electrochemical detection of hybridization in the waste area 109 (Waste 4).

This concludes the analysis process. In particular, in an electrochemical detection, the resulting signals can be read out electrically and processor-based evaluated according to a predetermined program.

The cartridge described in detail with channels and cavities with reference to FIG. 1 is made of a polymer material, such as polycarbonate, for example by injection molding. Initially, the card base body 101 is produced with structures that are open at the top, and the reagents are spotted into the initially open channels or cavities and then dried. The detection module is incorporated in a suitable manner in the cartridge, insbesonde ^¬ re glued. Finally, the channels and the ^{cavities are provided} , for example, with an elastic film as the top cover and are therefore closed for the intended use.

It is also possible that on the open card body 101 before the cover side completion and completion of the Cartridge certain special equipment, for example, as sealing materials and / or ventilation materials, aufge¬ be introduced.

The concrete measuring method was explained with reference to Figs. 11 to 23 for a specific case of DNA analysis of a sample of whole blood. Generally is provided which ne-described ^¬ Cartridge for DNA analysis of a hand and / or the Pro ^¬ protein analysis on the other hand use, wherein - as already mentioned above - an appropriate reader and zugehö ^¬ membered evaluation algorithms can be used as an aid. This results in defined operating methods which make the cartridge described in the examples only practical for decentralized use in the context of a medical "point of care" application.

Finally, especially for the DNA analysis, the integrated operating method of the cartridge described above in detail is summarized again as a combination or in the sequence of the individual substeps:

- Place the sample in cartridge

- Insert the cartridge in the reader

- Start of the fully automatic assay - Sample dosing via dosing line

- Washing the dosing line

- Dilute the sample and place in lysis channel

- Linger in the lysis channel

- Collection of the DNA bead complex by bead collectors in the PCR chamber

- Washing the DNA bead complex with water

- Close the PCR chamber

- Performing the PCR

- During the PCR: Fill the two ELISA reagent channels with water

- opening the PCR chamber

- Transport of the PCR product in detection chamber

- Hybridization in the detection chamber - Vent the two ELISA reagent channels

Filling and rinsing the detection chamber with ELISA reagent 1

- Fill and rinse the detection chamber with ELISA reagent 2

- Carrying out the electrochemical measurements.

In electrochemical measurements, the detection chamber is first flushed with an antibody solution carrying an enzyme label (ELISA reagent 1). Then, the rinsing of the detection chamber with enzyme substrate (ELISA reagent 2). The electrochemical measurements are carried out in a manner known per se at predeterminable, different temperatures and changeable flow rates of the enzyme-substrate solution.

For protein analysis, appropriate procedures are used, in which case the PCR does not apply.

Claims

claims

1. Arrangement for the integrated and automated DNA or protein analysis of a measurement sample in a disposable cartridge filled with dry reagents (card), having the following features for the automatic implementation of all measures necessary for the process analysis of individual partial processes in the cartidge:

in the cartridge (100) there is a system of microchannels and / or microcavities (101 to 131) for a microfluidic process technology,

- the microchannels or microcavities (101 to 131) wei ^¬ sen predetermined geometric structures to accommodate reagent, wherein - the reagents at specific points in the microchannels or microcavities (101 to 131) of the cartridge (100) in a are stored in a storage-stable form,

- There are means available to provide the dry-stored Reagen¬ zien for the respective sub-process in a suitable form, in particular as a liquid reagent.

2. Arrangement according to claim 1, characterized in that the structures for receiving the dry, storage-stable reagents reagents recesses (112, 132).

3. Arrangement according to claim 2, characterized in that the depressions have at least one step with step heights of 10 to 500 microns.

4. Arrangement according to claim 2, characterized in that the depressions have an extension of about 1 mm and a depth of about 100 microns.

5. Arrangement according to claim 3, characterized in that the recesses are Cylindrical, wherein the expansion of the Diameter represents.

6. Arrangement according to one of the preceding claims, characterized in that in the microchannels or in the micro cavities (101 to 131) introduced reagents have the following properties: - There are dry substances with negligible

Vapor pressure, which are stable at room temperature, so that the properties for a ZellaufSluluss or a PCR or for the detection of biochemical sizes are retained.

7. Arrangement according to one of the preceding claims, characterized in that mixtures of the respective substance with excipients form thin films which adhere to the walls.

8. Arrangement according to claim 7, characterized in that in parts of the microchannels or microcavities (101 to 131) a ^¬ brought substances or mixtures with thin paraffin layers are covered waterproof.

9. Arrangement according to one of the preceding claims, characterized in that in parts of the microchannels or micro ^¬ cavities (101 to 131) introduced substances have DNA- or protein-binding properties.

10. Arrangement according to one of the preceding claims, da¬ characterized in that the introduced into parts of the microchannels or microcavities (101 to 131) substances are magnetic beads with specific binding properties.

11. Arrangement according to claim 10, characterized in that the magnetic beads are coated with antibodies.

12. Arrangement according to claim 10, characterized in that the magnetic beads are coated with DNA-binding substances.

13. Arrangement according to one of the preceding claims, wherein equally lysis reagents and magnetic beads are present, characterized in that the lysis reagents and the magnetic beads are contained in a single dry matrix.

14. Arrangement according to one of the preceding claims, da¬ characterized in that a so-called DNA ELISA assay or protein ELISA assay (130, 131) is feasible, being used as reagents for the ELISA assay (130, 131) a label enzyme (reagent 1) and an enzyme substrate (reagent 2) are present.

15. Arrangement according to one of the preceding claims, da¬ characterized in that a detection module (130) for the electrical detection of the hybridization processes is present.

16. The arrangement according to claim 15, characterized in that the detection module (130) consists of a precious metal / plastic composite.

17. Arrangement according to claim 15, characterized in that the detection module consists of a semiconductor-processed silicon chip with noble metal electrodes.

18. Arrangement according to claim 15, characterized in that electrochemical, magnetic and / or piezoelectric measuring methods are used as means for the electrical detection.

19. Arrangement according to one of the preceding claims, da¬ characterized in that the cartridge (100) has an input port (102) for a whole blood sample.

20. Arrangement according to one of the preceding claims, character- ized in that means (103 to 103 ''') to Zu drove by water, esp. An influx, is present.

21. Arrangement according to claim 20, characterized in that an inflow port for connection to an external water supply is present.

Is that the inflow port is to an integrated water reservoir ^¬ closed 22. The arrangement according to claim 20, characterized.

23. Arrangement according to one of the preceding claims, da¬ characterized in that the microchannels or Mikrokavitä- th (101 to 131) are filled with dry buffer substances of defined ionic strength after water supply.

24. Arrangement according to one of the preceding claims, da¬ characterized in that means for mixing whole blood ^¬ samples and water or the buffer solution are present.

25. Arrangement according to one of the preceding claims, da¬ characterized in that means for flowing through the microsphere coated with lysis bead reagent or microcavity with blood or with blood-water or blood-buffer mixture are present ,

26. Arrangement according to one of the preceding claims, da¬ characterized in that means for generating a Mag ^¬ netfeldes for the purpose of fixing the DNA / magnetic bead or protein in-magnetic bead complex are present.

27. Arrangement according to one of the preceding claims, da¬ characterized in that means for generating a Mag ^¬ netfeldes for the purpose of fixing the DNA / magnetic bead complex in a PCR cavity are present.

28. Arrangement according to claim 21, characterized in that means for closing the PCR cavity present are .

29. Arrangement according to one of the preceding claims, da¬ characterized in that means for thermocycling of the sample are present.

30. Arrangement according to one of the preceding claims, da¬ characterized in that in the cartridge (card) means for storing spent sample material and / or used reagents are present.

31. Arrangement according to claim 30, characterized in that the means for storing spent sample material and / or spent reagents form waste reservoir.

32. Arrangement according to one of the preceding claims, da¬ characterized in that means for germ-proof, particle or cell-free venting of the waste reservoirs are present.

33. Arrangement according to one of the preceding claims, da¬ characterized in that means for fixing the cartridge are present in a read-out device.

34. A method for producing a cartridge used in the arrangement according to claim 1 or any one of claims 2 to 33, comprising the following method steps:

- A cartridge body with channels and / or cavities is made of polymer, - reagents are spotted in the open channels and dried there,

- The channels and / or cavities are closed by a film ver¬.

35. A manufacturing method according to claim 34, characterized gekenn¬ characterized in that the card body by Spritzgusstech- nik is produced.

36. A manufacturing method according to claim 34, characterized gekenn¬ characterized in that special materials, such as sealing membranes, venting membranes od. Like., Are applied to the karten body.

37. A manufacturing method according to claim 34, characterized gekenn¬ characterized in that before the sealing of the card body a detection module tion with measuring means introduced, in particular ^¬ sticks is.

38. The method of operation for DNA analysis in an assembly according to claim 1 or any one of claims 2 to 33, with the following measures:

- placing the sample in the cartridge,

- insert the cartridge into the reader,

- Start the fully automatic assay.

39. Operating method according to claim 38, with the following steps in the operation of the fully automatic assay:

A sample dosing takes place via a metering section,

the dosing line is washed,

the test sample is diluted and introduced into the lysis channel,

- by lingering in the lysis channel a ZellaufSchluss,

- the formed DNA-bead complex is brought by a liquid keitsstrom ^¬ into the PCR chamber and held by a bead collector in the PCR chamber - it is carried out by washing the DNA-bead complex with water,

- the PCR chamber is closed,

- the PCR is carried out

after completion of the PCR, the PCR product is transported into the detection chamber - hybridization processes with specific capture probes take place in the detection chamber,

- There is the flushing of the detection chamber with Markie¬ tion enzyme - There is the flushing of the detection chamber with enzyme substrate

- The electrochemical measurements are carried out

the electrochemical measurements are carried out at different temperatures and different flow velocities

Enzyme substrate solution performed.

40. Operating method according to claim 39, characterized gekennzeich¬ net, that during the PCR, the ELISA reagent channels are filled with water.

41. Operating method according to claim 39, characterized in that the two ELISA channels are vented gekennzeich¬ net after hybridization, that subsequently the detection chamber initially free of gas bubbles purged with the first ELISA reagent and then gas bubbles ge with the second ELISA reagent ^¬ is rinsed, and that subsequently the electrochemical Mes ^{¬ solution} is performed.

42. Operating method for a protein analysis in a An¬ arrangement according to claim 1 or one of claims 2 to 33, with the following measures:

- placing the sample in the cartridge,

- insert the cartridge into the reader, - start the fully automatic assay.

43. Operating method according to claim 42, with the following steps in the operation of the fully automatic assay:

- A dosing is done via a dosing, - the dosing is washed,

- The sample is diluted and transported by a liquid ^¬ stream in the detection chamber

In the detection chamber, binding processes take place between the proteins of the test sample and specific capture antibodies or catcher proteins; the detection chamber is flushed with egg an antibody solution bearing an enzyme label (ELISA reagent 1),

- The detection chamber is flushed with enzyme substrate (ELISA reagent 2) - the electrochemical measurements are carried out.

44. Operating method according to claim 43, characterized gekennzeich¬ net, that after the hybridization, the two ELISA channels are vented, then subsequently flushed the detection chamber to ^¬ next with the first ELISA reagent gas bubble-free and then with the second ELISA reagent gas bubble-free ge ^¬ is rinsed, and that subsequently the electrochemical Mes ^{¬ solution} is performed.