WO2021219533A1

WO2021219533A1 - Cartridge, system and method for amplification of at least one analyte

Info

Publication number: WO2021219533A1
Application number: PCT/EP2021/060806
Authority: WO
Inventors: Axel Niemeyer
Original assignee: Boehringer Ingelheim Vetmedica Gmbh
Priority date: 2020-04-29
Filing date: 2021-04-26
Publication date: 2021-11-04

Abstract

A cartridge, heating element, system and method for amplification of at least one nucleic acid of an in particular biological sample is proposed, wherein the heating element is inductively heated and arranged completely in a reaction cavity to contact directly the sample in order to heat it partially.

Description

Cartridge, system and method for amplification of at least one analyte

The present invention relates to a cartridge for amplification of at least one analyte, preferably a nucleic acid sequence, of an in particular biological sample according to the preamble of claim 1 , a heating element for amplification of at least one nucleic acid sequence according to the preamble of claim 120, a system for amplification of at least one analyte, preferably a nucleic acid sequence, of an in particular biological sample according to the preamble of claim 30, and a method for amplification of at least one analyte, preferably a nucleic acid sequence, of an in particular biological sample according to the preamble of claim 34.

Preferably, the present invention deals with the amplification of at least one nucleic acid, in particular by polymerase chain reaction (PCR), wherein part of the sample is heated in a reaction cavity by a heating element. If only part of the sample is heated a faster amplification or PCR can be realized.

WO 2018/073435 A1 discloses a method for the amplification of nucleic acids by PCR, wherein a wire is led through a reaction cavity and heats electrically by resistive heating only part of the sample or liquid contained in the reaction cavity. This allows fast temperature changes and, thus, fast PCR. Potentially problematic is the electri cal contact of the wire. Further, the fluid-tight integration of the wire into the reaction cavity is challenging.

In particular, the present invention deals with analyzing and testing a preferably bio logical sample, in particular from a human or animal, mostly preferred for analytics and diagnostics, e.g. with regard to the presence of diseases and/or pathogens and/or for determining blood counts, antibodies, hormones, steroids or the like.

Therefore, the present invention is in particular within the field of bioanalytics. A food sample, environmental sample or another sample may optionally also be tested, in particular for environmental analytics or food safety and/or for detecting other sub stances.

Preferably, by means of the present invention, at least one analyte (target analyte) of a sample can be determined, identified or detected. In particular, the sample can be tested for qualitatively or quantitatively determining at least one analyte, e.g. in order to detect or identify a disease and/or pathogen. Within the meaning of the present invention, analytes are in particular nucleic-acid sequences, in particular DNA sequences and/or RNA sequences, and/or proteins, in particular antigens and/or antibodies. In particular, by means of the present inven- tion, nucleic-acid sequences or proteins can be determined, identified or detected as the analytes of a sample. Mostly preferred, the present invention deals with systems, devices and other apparatuses for carrying out a nucleic-acid assay for detecting or identifying a nucleic-acid sequence or a protein assay for detecting or identifying a protein.

The present invention deals in particular with what are known as point-of-care sys tems, e.g. mobile systems/devices and other mobile apparatuses, and deals with methods for carrying out tests on a sample at the sampling site and/or independently or away from a central laboratory or the like. Preferably, point-of-care systems can be operated autonomously and/or independently of a mains network for supplying electrical power.

WO 2018/065110 A1 discloses an analyzer and a method for testing a biological sample, wherein the analyzer comprises a receiving unit for receiving, positioning and holding the cartridge and a connection unit for mechanically, electrically, ther mally and/or fluidically connecting the cartridge. The receiving unit can be moved relative to the connection unit in order to hold the cartridge in a clamped manner between the receiving unit and the connection unit and to heat reaction cavities from opposite sides by means of Peltier elements for amplifying analytes, in particular nucleic acid sequences, of the sample in the reaction cavities by PCR.

PCR is relatively time-consuming because the heating and cooling of the sample has to repeated several times and requires for each heating cooling step a relatively long time.

US 2011/0008797 A1 discloses a cartridge which is provided with functionalized magnetic beads or particles of up to 10 pm, which are inductively heated. Such beads or particles are not suitable for pre-insertion in a flow-through cavity with inlet and outlet on opposite sides.

US 2018/0136246 A1 discloses a system for inductive heating of a sample for ana lyte detection. The system comprises at least one sample container and a receptacle with an induction coil for receiving the sample container. The sample container com prises an interior space with only one opening at the upper end, wherein the heating element including an electrically conductive portion is arranged within the interior space. The heating element is rod-shaped or forms a hollow cylinder or ring. The sample container is placed within the receptacle so that the central axis of the induc tion coil extends through the sample container. A cartridge having a fluidic system and fluidically connected cavity is not disclosed. Further, the heating element is not optimized for amplification of at least one analyte. US 2013/0244241 A1 discloses an analyzer for receiving a cartridge for receiving a sample to be analyzed. The cartridge comprises a movable transfer module with a transfer chamber for transferring liquids between various chambers of the cartridge. The transfer chamber may be provided with a stirring element in the form of a mag netic stir bar for mixing the contents in the transfer chamber. The cartridge may com- prise one or more reaction chambers for performing a PCR. The reaction chambers are covered by a thin film for allowing easy heating and/or cooling of the contents via an external source. In particular, the films may be in contact with a surface that is thermally controlled by a thermoelectric device, resistive heater or forced air. Thus, any conductive heating for PCR is not disclosed.

Object of the present invention is to provide a cartridge, a heating element, a system and a method for amplification of at least one analyte, preferably a nucleic acid se quence, of an in particular biological sample, which allow or facilitate a rapid, efficient, reliable, comprehensive and/or precise amplification and which are easy to use, manufacture and assemble.

The above object is achieved by a cartridge according to claim 1 , by a heating ele ment according to claim 20, by system according to claim 30 or by a method accord ing to claim 34. Advantage developments are subject of the dependent claims.

The proposed cartridge preferably comprises a reaction cavity for receiving the sam ple and comprises at least one or only one inductively heatable heating element. Preferably, the heating element is arranged completely in the reaction cavity to con tact directly the sample in order to heat it. Preferably, the heating element is induc- tively heatable by an inductor adjacent to the reaction cavity. Thus, any electrical connection of the heating element can be avoided, which would complicate liquid- tight sealing of the reaction cavity. Thus, integration of the heating element into to the reaction cavity and assembly of a respective cartridge are facilitated.

In particular, the heating element can be produced independently from other parts of the cartridge. Preferably, the heating element can be pretreated, e.g. functionalized, and/or provided with one or more dried reagents and, then, placed into the reaction cavity before closing the reaction cavity or a side of the cartridge. This facilitates production and assembly. Preferably, the heating element can be completely surrounded by the sample. This allows optimized heat transfer and, thus, quicker heating cycles for faster amplifica tion or PCR.

Preferably, the sample is heated by the heating element only or primarily in the region near and/or close to the heating element. In particular, it is not necessary to heat the whole sample received in the reaction cavity for the amplification of at least one nu cleic acid.

This can reduce drastically the time needed for temperature cycling and, thus, for the amplification process as the cooling of the sample can be promoted by the unheated sample part in the reaction cavity and by the surrounding walls of the reaction cavity. Therefore, a very rapid cooling and, thus, thermal cycling of the heating-up zone of the sample close to the heating element can be reached. Thus, the unheated passive volume can preferably cool the heated part of the sam ple. The unheated passive volume is many times greater than the volume of the sample heated up by the heating element. Due to that fact the invention leads in particular to an amplification process which is carried out more rapidly. In particular, the limitation of the time needed for the amplification of the nucleic acid(s) is by- passed by using the heating element.

Preferably, the heating element is a helically formed wire, in particular a substantially, preferably closed, circular and/or elliptically helically formed wire. In particular, the heating element is ring-like, foil-like or wire-like. Such a design enables in particular to provide a large surface area of the heating element for the amplification of nucleic acid(s) and/or for heating the part of the sample close to the outer surface area of the heating element. Especially preferred, the heating element forms a closed conductor loop or coil for high energy absorption and/or heating power. According to a preferred embodiment of the present invention, the heating element is loosely arranged in the reaction cavity. This allows very simple assembly.

Preferably, the heating element can be designed as an insert component which is inserted or dropped into the reaction cavity before closing the reaction cavity during assembly of the cartridge. Advantageously, any particular alignment or positioning of the heating element is not necessary. This facilitates assembly.

In particular, the sample received in the reaction cavity is a liquid solution or suspen sion which contains the nucleic acid sequence(s) to be amplified. It generally can also contain besides the nucleic acid sequence(s) to be amplified complements and/or other constituent parts or reagents, for example polymerase(s), dNTBs and/or salts, which can be suspended or dissolved and/or provides also by the heating ele ment, e.g. as dried reagents that are dissolved, e.g. by the sample. The term "cartridge" is preferably understood to mean a structural apparatus or unit designed to receive, to store, to physically, chemically and/or biologically treat and/or prepare and/or to measure a sample, preferably in order to make it possible to detect, identify or determine at least one analyte, in particular a protein and/or a nucleic-acid sequence, of the sample.

In particular, within the meaning of the present invention, a cartridge is designed to be at least substantially planar and/or card-like, in particular is designed as a (mi cro )fluidic card and/or is designed as a main body or container that can preferably be closed and/or said cartridge can be inserted and/or plugged into an analyzer (analysis device) when it contains the sample.

A cartridge within the meaning of the present invention preferably comprises a fluid system having a plurality of channels, cavities and/or valves for controlling the flow through the channels and/or cavities.

The proposed heating element is functionalized with oligonucleotides, preferably pri mers, and/or other reagents for the amplification and/or is provided with one or more preferably dried reagents for the amplification and can be used as an insert that can be perfectly manufactured and prepared, in particular functionalized and/or provided with reagents, prior to placing or integrating the heating element into the respectively cavity. This facilitates manufacture and assembly significantly.

Further, the proposed heating element is made at least partly of an electroconductive material, preferably a ferromagnetic material and/or forms a closed conductor loop or coil and/or an areal or flat arrangement. This supports high energy absorption and/or heating power while creating a relatively large surface for heating the closed surrounding essentially.

The proposed system of a cavity with an inductively heatable heating element and the associated inductor for inductively heating the heating element for the amplifica tion of at least one analyte or nucleic acid sequence allows very fast thermic cycles and, thus, very fast amplification, in particular PCR, while a simple assembly and construction are possibly.

Preferably, the system comprises a cartridge with the heating element as described above and analyzer with the inductor for receiving and/or controlling the cartridge and for inductively heating the heating element and/or controlling the amplification or thermic cycling.

In the context of the present invention, the term "analyzer" is preferably understood to refer to a preferably mobile instrument/apparatus, which is designed to chemically, biologically and/or physically tests and/or analyze a sample or a component thereof, preferably in and/or by means of a cartridge containing the sample. The analyzer preferably controls the testing of the sample in and/or by means of the cartridge. In order to carry out the test, the cartridge can be connected to, in particular received by, the analyzer, as already mentioned.

Furthermore, the proposed method for amplification of at least one analyte or nucleic acid sequence of an in particular biological sample uses at least one inductively heat able heating element arranged completely in a reaction cavity and heating the sam ple, preferably only partly and/or close to the heating element. This leads to similar advantages as described above. The above-mentioned aspects and features of the present invention and the aspects and features of the present invention that will become apparent from the claims and the following description can, in principle, be implemented independently from one another, but also in any combination or order.

Other aspects, advantages, features and properties of the present invention will be come apparent from the claims and the following description of a preferred embodi ment with reference to the drawings, in which:

Fig. 1 is a schematic view of a proposed system with an analyzer and a pro posed cartridge received in the analyzer;

Fig. 2 is a schematic perspective front view of the cartridge with proposed heating elements;

Fig. 3 is a schematic perspective rearview of the cartridge; Fig. 4 is a schematic perspective view of the analyzer in the open state; Fig. 5 is a schematic sectional view of the analyzer, showing the analyzer in the open position;

Fig. 6 is a schematic view of a proposed heating element; Fig. 7 is a schematic cross-section of the heating element; and

Fig. 8 is a schematic sectional view of the heating element received in a re action cavity of the cartridge. In the Figures, the same reference signs are used for the same or similar parts and components, resulting in corresponding or comparable properties, features and ad vantages, even if these are not repeatedly described.

Fig. 1 is a highly schematic view of a proposed system 10 comprising an apparatus or cartridge 100 for testing an in particular biological sample P and/or for amplifying at least one analyte or nucleic acid sequence of the sample P. Preferably, the system 10 comprises an analyzer 200 in particular for receiving the cartridge 100 and/or controlling the amplification and/or testing in the cartridge 100.

Fig. 2 is a perspective front view of the cartridge 100 showing its front 100A and Fig. 3 is a perspective rear view thereof, showing its back 100B.

The apparatus or cartridge 100 in particular forms a handheld unit, hereinafter re ferred to as cartridge 100. The term "sample" is preferably understood to refer to a sample material that is to be tested and which is in particular taken from a human or animal. Preferably, within the meaning of the present invention, a sample is a fluid, such as saliva, blood, urine or another liquid, preferably from a human or animal, or a component thereof. Within the meaning of the present invention, a sample may be pre-treated or pre pared if necessary, or may come directly from a human or animal or the like. A food sample, environmental sample or another sample may optionally also be tested, in particular for environmental analytics, food safety and/or for detecting other sub stances, preferably natural substances, but also biological or chemical warfare agents, poisons or the like.

A sample within the meaning of the present invention preferably contains one or more analytes, it preferably being possible for the analytes to be identified or detected, in particular qualitatively and/or quantitatively determined. Preferably, within the mean- ing of the present invention, a sample has target nucleic-acid sequences as analytes, in particular target DNA sequences and/or target RNA sequences, and/or target pro teins as the analytes, in particular target antigens and/or target antibodies. Prefera bly, at least one disease and/or pathogen can be detected or identified in the sample P by qualitatively and/or quantitatively determining the analytes.

Preferably, the analyzer 200 controls an amplification, in particular by PCR, of one or more analytes for the testing of the sample P and preferably the testing, in partic ular in or on the cartridge 100, and/or is used to evaluate the testing and/or to collect, to process and/or to store measured values from the test.

By means of the analyzer 200 and/or by means of the cartridge 100 and/or by means of the method for testing the sample P, an analyte or a plurality of analytes of the sample P can preferably be determined, identified or detected, in particular not only qualitatively, but also quantitatively.

Therefore, the sample P can in particular be tested for qualitatively and/or quantita- tively determining at least one analyte, e.g. in order to detect or identify a disease and/or a pathogen or to determine other values, which are important for diagnostics, for example.

Particularly preferably, a molecular-biological test is made possible by means of the analyzer 200 and/or by means of the cartridge 100.

Particularly preferably, a nucleic-acid assay for detecting or identifying a target nu cleic-acid sequence, in particular a target DNA sequence and/or a target RNA se quence, and/or a protein assay for detecting or identifying a target protein, in partic- ular a target antigen and/or target antibody, are made possible or are carried out.

The term "assay" is preferably understood to mean an in particular molecular-biolog ical test for detecting or identifying at least one analyte in a sample P. In particular, at least one analyte in a sample P can be qualitatively or quantitatively detected or identified by means of an assay or by carrying out an assay. A plurality of method steps is preferably required to (fully) carry out an assay. Preferably, within the mean ing of the present invention, when carrying out an assay, a sample P is pretreated with one or more reagents and the pretreated sample P is tested, in particular at least one analyte in the sample P being detected or identified. Within the meaning of the present invention, an assay is in particular an immunoassay or protein assay for de tecting or identifying a target protein, in particular a target antigen and/or target anti body, and/or a nucleic-acid assay for detecting or identifying a target nucleic-acid sequence, in particular a target DNA sequence and/or target RNA sequence. Preferably, the sample P or individual components of the sample P or analyte can be amplified if necessary, in particular by means of PCR, and tested, detected or identified in the analysis device / analyzer 200 or in the cartridge 100, and/or for the purpose of carrying out the nucleic-acid assay. Preferably, amplification products of the analyte or analytes are thus produced.

In the following, further details are first given on a preferred construction of the car tridge 100, with features of the cartridge 100 preferably also directly representing features of the system 10, the analyzer 200 or the method for amplification of at least one analyte, in particular even without any further explicit explanation.

The cartridge 100 is preferably at least substantially planar, flat, plate-shaped and/or card-like.

The cartridge 100 preferably comprises an in particular at least substantially planar, flat, plate-shaped and/or card-like main body / support 101 , the main body or support 101 in particular being made of and/or injection-moulded from plastic material, in particular polypropylene.

The cartridge 100 preferably comprises two flat sides 100A, 100B. In particular, the front 100A of the cartridge 100 and the back 100B of the cartridge 100 are each a flat side of the in particular planar and/or card-like cartridge 100.

The cartridge 100 preferably comprises at least one film/cover 102 for covering the main body 101 and/or cavities and/or channels formed therein, at least partially, in particular on the front 100A, and/or for forming valves or the like. The cartridge 100 and/or its main body 101 , in particular together with the cover 102, preferably forms and/or comprises a fluidic system 103, hereinafter referred to as fluid system 103.

The cartridge 100, the main body 101 and/or the fluid system 103 are/is preferably at least substantially vertically oriented during the operation/test and/or in the test/op erating position and/or when being inserted in the analyzer 200, as shown schemat ically in Fig. 1. In particular, the surface extension or main plane H of the cartridge 100 extends at least substantially vertically in the test/operating position. The cartridge 100, in particular its fluid system 103, preferably comprises a plurality of cavities, in particular at least one receiving cavity 104 for receiving/introducing the sample P, at least one metering cavity 105, at least one intermediate cavity 106, at least one mixing cavity 107, at least one storage cavity 108, at least one reaction cavity 109, at least one intermediate temperature-control cavity 110 and/or at least one collection cavity 111 , a plurality of cavities preferably being fluidically intercon nected in particular by a plurality of channels. Within the meaning of the present invention, channels are preferably elongate forms for conducting a fluid in a main flow direction, the forms preferably being closed trans versely, in particular perpendicularly, to the main flow direction and/or longitudinal extension, preferably on all sides.

In particular, the main body 101 comprises elongate notches, recesses, depressions or the like, which are closed at the side by the cover 102 and form channels within the meaning of the present invention. Within the meaning of the present invention, cavities or chambers are preferably formed by recesses, depressions or the like in the cartridge 100 or main body 101 , which are closed or covered by the cover 102, in particular at the side. The volume or space enclosed by each cavity is preferably fluidically linked, in particular to the fluid system 103, by means of channels.

In particular, within the meaning of the present invention, a cavity comprises at least two openings for the inflow and/or outflow of fluids.

Within the meaning of the present invention, cavities preferably have a larger diam- eter and/or flow cross section than channels, preferably by at least a factor of 2, 3 or 4. In principle, however, cavities may in some cases also be elongate, in a similar manner to channels.

Preferably, several or all of the cavities are vertically oriented and/or are oriented such that fluid can flow through the cavities at least substantially vertically in the normal operating position of the cartridge 100.

Particularly preferably, several or all of the cavities, in particular the receiving cavity 104, the intermediate cavity/cavities 106, the mixing cavity 107, the storage cav- ity/cavities 108 and/or the reaction cavity/cavities 109, are elongate, the longitudinal extension of the cavities preferably extending at least substantially vertically, and/or in parallel with gravity G in the normal operating position of the cartridge 100.

Preferably, the inlet of several or all of the cavities is at the top in the normal operating position of the cartridge 100 and the outlet of several or all of the cavities is at the bottom in the normal operating position of the cartridge 100, in particular such that fluid can flow through or drain from some or all of the cavities, in particular the storage cavity/cavities 108, from the top to the bottom in the normal operating position and/or a fluid located in the cavities, in particular the storage cavity/cavities 108, can be removed and/or pumped out at the bottom. In this way, bubble formation and/or foaming of the fluids located in the cavities can be prevented. In particular, this pre- vents a gas, in particular air, from being conveyed out of the cavities.

The cartridge 100 and/or the fluid system 103 preferably comprises at least one pump apparatus 112 and/or at least one sensor arrangement / apparatus 113. In the example shown, the cartridge 100 or the fluid system 103 preferably comprises a plurality of intermediate cavities 106, a plurality of storage cavities 108 and/or a plurality of reaction cavities 109, which can preferably be loaded separately from one another, in particular a first reaction cavity 109, a second reaction cavity 109 and an optional third reaction cavity 109, as can be seen in Fig. 2.

In the initial state of the cartridge 100 or when at the factory, the storage cavities 108 are preferably filled at least in part, in particular with a liquid such as a reagent, sol vent or wash buffer. The reaction cavity/cavities 109 is/are preferably designed to allow a substance lo cated in the reaction cavity 109 to react when an assay is being carried out.

The reaction cavity/cavities 109 is/are used in particular to carry out an amplification reaction, in particular PCR, or several, preferably different, amplification reactions, in particular PCRs. It is preferable to carry out several, preferably different, PCRs, i.e. PCRs having different primer combinations or primer pairs, in parallel and/or inde pendently and/or in different reaction cavities 109.

To carry out the nucleic-acid assay, preferably target nucleic-acid sequences, as an- alytes A of the sample P, are amplified in the reaction cavity/cavities 109 by means of an amplification reaction, in particular in order to produce amplification products for the subsequent detection in the sensor arrangement or sensor apparatus 113.

Within the meaning of the present invention, the amplification or also-called amplifi cation reaction is an in particular molecular-biological reaction in which an analyte, in particular a target nucleic-acid sequence, is amplified/copied and/or in which am plification products, in particular nucleic-acid products, of an analyte are produced. Particularly preferably, PCRs are amplification reactions within the meaning of the present invention.

"PCR" stands for polymerase chain reaction and is a molecular-biological method by means of which certain analytes, in particular portions of RNA or RNA sequences or DNA or DNA sequences, of a sample P are amplified, preferably in several cycles, using polymerases or enzymes, in particular in order to subsequently test and/or de tect the amplification products or nucleic-acid products. If RNA is intended to be tested and/or amplified, before the PCR is carried out, a cDNA is produced starting from the RNA, in particular using reverse transcriptase. The cDNA is used as a tem plate for the subsequent PCR.

Preferably, during a PCR, a sample P is first denatured by the addition of heat in order to separate the strands of DNA or cDNA. Preferably, primers or nucleotides are then deposited on the individual separated strands of DNA or cDNA, and a de sired DNA or cDNA sequence is replicated by means of polymerase and/or the miss ing strand is replaced by means of polymerase. This process is preferably repeated in a plurality of cycles (each cycle requiring a temperature change by heating and cooling) until the desired quantity of the DNA or cDNA sequence, i.e. analyte, is available.

For the PCR, marker primers are preferably used, i.e. primers which (additionally) produce a marker or a label L, in particular biotin, on the amplified analyte or ampli fication product. This allows or facilitates detection. Preferably, the primers used are biotinylated and/or comprise or form in particular covalently bonded biotin as the la bel L (schematically shown in Fig. 7).

Preferably, the cartridge 100 comprises at least one inductively heatable heating el ement 165 and/or one or multiple or all cavities 109 are provided with an inductively heatable heating element 165, as schematically indicated in Fig. 2, for thermo cycling and/or the amplification process. The heating element 165 will be described later in detail.

The amplification products, target nucleic-acid sequences and/or other portions of the sample P produced in the one or more reaction cavities 109 can be detected and/or are preferably conducted or fed to the connected sensor arrangement or sen sor apparatus 113, in particular by means of the pump apparatus 112. The sensor arrangement or sensor apparatus 113 is used in particular for detecting, particularly preferably qualitatively and/or quantitatively determining, the analyte or analytes of the sample P, mostly preferred the target nucleic-acid sequences and/or target proteins as the analytes. Alternatively or additionally, however, other values may also be collected and/or determined.

The sensor apparatus 113 preferably comprises a sensor array (not shown) in order to determine or detect in particular a plurality of analytes.

In particular, the sensor apparatus 113 or sensor array comprises capture molecules (not shown) in order to bond analytes and/or amplification products and subsequently detect, identify or determine said analytes and/or amplification products in a detec tion process.

Mostly preferred, electrochemical detection is carried out.

The cartridge 100, the main body 101 and/or the fluid system 103 preferably com prise a plurality of channels 114 and/or valves 115, as shown in Fig. 2.

By means of the channels 114 and/or valves 115, the cavities 104 to 111 , the pump apparatus 112 and/or the sensor apparatus 113 can be temporarily and/or perma nently fluidically interconnected and/or fluidically separated from one another, as re quired and/or optionally or selectively, in particular such that they are controlled by the analyzer 200.

The cavities 104 to 111 are preferably each fluidically linked or interconnected by a plurality of channels 114. In particular, each cavity is linked or connected by at least two associated channels 114, such that the fluid can fill, flow through and/or drain from the respective cavities as required.

The fluid transport or the fluid system 103 is preferably not or not exclusively based on capillary forces, but is preferably essentially based on the effects of gravity and/or pumping forces, compressive forces and/or suction forces that arise and/or that are generated by the pump or pump apparatus 112. Mostly preferred, the flow or transport of fluids and the metering are controlled by accordingly opening and closing the valves 115 and/or by accordingly operating the pump apparatus 112, in particular by means of the analyzer 200, in particular its pump drive 202.

Preferably, at least one valve 115 is assigned to each cavity, the pump apparatus 112 and/or the sensor apparatus 113 and/or is arranged upstream of the respective inlets and/or downstream of the respective outlets. Preferably, by actuation of the assigned valves 115, the cavities 104 to 111 or se quences of cavities 104 to 111 , can be selectively released and/or fluid can selec tively flow therethrough, and/or the cavities 104 to 111 can be fluidically connected to the fluid system 103 and/or to other cavities. In particular, the valves 115 are formed by the main body 101 and the film/cover 102 and/or are formed therewith and/or are formed in another manner, e.g. by additional layers, depressions or the like.

Preferably, one or more valves 115A are - preferably tightly - closed initially and/or in the delivery state of the cartridge 100, in particular in order to seal liquids or liquid reagents F, located in the storage cavities 108, and/or the fluid system 103 from the open receiving cavity 104 and/or in a storage-stable manner. Hereinafter, these valves 115A are referred to as initially closed valves 115A. Preferably, an initially closed valve 115A is arranged upstream and downstream of each storage cavity 108. Said valves 115Aare preferably (only) opened, in particular automatically and/or by means of the analyzer 200, when the cartridge 100 is actually being used, in particular for the first time, and/or during or after inserting the cartridge 100 into the analyzer 200 and/or for carrying out the assay.

The initially closed valves 115A assigned to the receiving cavity 104 seal the fluid system 103 and/or the cartridge 100 in particular fluidically and/or in a gas-tight man ner, preferably until the sample P is introduced and/or the receiving cavity 104 is closed.

As an alternative or in addition to the initially closed valves 115A, one or more valves 115B are preferably provided which are open / not closed initially/normally and/or in the delivery state of the cartridge 100 and/or in an inoperative/initial position/state and/or when the cartridge 100 is not inserted into the analyzer 200. These valves 115B are used in particular to control the flows of fluid during the test and/or are referred to as initially/normally open valves 115B.

Preferably, the normally open valves 115B can (only) be closed by actuation, mostly preferred by means of the analyzer 200.

The cartridge 100 is preferably designed as a microfluidic card and/or the fluid sys- tern 103 is preferably designed as a microfluidic system and/or the reaction cavity 109 is preferably a microfluidic chamber.

In the present invention, the term "microfluidic" is preferably understood to mean that the respective volumes of the individual cavities, some of the cavities or all of the cavities 104 to 111 and/or channels 114 are, separately or cumulatively, less than 5 ml or 2 ml, preferably less than 1 ml or 800 pi, in particular less than 600 mI or 300 mI, mostly preferred less than 200 mI or 100 mI.

Preferably, a sample P having a maximum volume of 5 ml, 2 ml or 1 ml can be intro- duced into the cartridge 100 and/or the fluid system 103, in particular the receiving cavity 104.

For example, the sample P may be introduced into the receiving cavity 104 and/or cartridge 100 by means of a pipette, syringe or other instrument.

Preferably, (all) reagents and liquids required for the test, the amplification, the de tection process and/or for other purposes are provided in the cartridge 100, i.e. intro duced before the test, mostly preferred in liquid form as liquids or liquid reagents F and/or in dry form as dry reagents S, as indicated in the schematic view according to Fig. 2.

Furthermore, also (all) other liquids F required for the test, the amplification, the de tection process and/or for other purposes, in particular in the form of a wash buffer, a solvent for dry reagents S and/or a substrate, e.g. in order to form detection mole cules and/or a redox system, are preferably provided in the cartridge 100, i.e. intro duced before use, in particular before delivery. The cartridge 100 preferably contains all the reagents and liquids required for pre treating the sample P and/or for carrying out the test or assay, in particular for carry ing out one or more amplification reactions or PCRs. Therefore, it is preferably only necessary to receive the optionally pre-treated sample P.

The cartridge 100, the fluid system 103 and/or the channels 114 preferably comprise sensor portions 116 or other apparatuses for detecting liquid fronts and/or flows of fluid. It is noted that in Figs. 2 and 3 various components, such as the channels 114, the valves 115, in particular the initially closed valves 115A and the normally open valves 115B, and the sensor portions 116 are, for reasons of clarity, only labelled in some cases. However, the same symbols are used in Figs 2 and 3, respectively, for each of these components.

As shown in Fig. 3, the sensor apparatus 113 preferably comprises electrical con tacts 113E for electrically connecting the cartridge 100 and/or sensor apparatus 113.

The contacts 113E are arranged in particular on the flat side and/or back and/or around a central region 113H.

The cartridge 100 and/or the main body 101 preferably has, in the region of the re action cavity/cavities 109, optionally a region of reduced wall thickness, a weakened portion or a depression 101 E, preferably for heating the cavity/cavities 109.

The receiving cavity 104 can be closed after the sample P has been received. The cartridge 100 preferably comprises a closure element 130 for this purpose.

In particular, the receiving cavity 104 can be closed in a liquid-tight and particularly preferably also gas-tight manner by the closure element 130. In particular, a closed fluid circuit can thus be formed, with the receiving cavity 104 being included. In par ticular, once the assigned valves 115A at the inlet, outlet and/or an intermediate con nection of the receiving cavity 104 have been opened, the receiving cavity 104 thus forms part of the fluid system 103 of the cartridge 100, wherein the fluid system is preferably closed or can be closed by the closure element 130. Once the sample P has been introduced into the receiving cavity 104 and said cavity has been closed, in particular by means of the closure element 130, the cartridge 100 can be inserted into and/or received in the proposed analyzer 200 in order to test the sample P, as shown in Figs. 1 , 4 and 5.

The analyzer 200 preferably comprises a preferably movable mount or receptacle 201 for mounting and/or receiving the cartridge 100. Preferably, the receptacle 201 can be moved up and down in order to eject and receive the cartridge 100, respec tively.

Preferably, the cartridge 100 and/or the fluid system 103 is fluidically, in particular hydraulically, separated or isolated from the analyzer 200. In particular, the cartridge 100 forms a preferably independent and in particular closed or sealed fluidic or hy draulic system 103 for the sample P and the reagents and other liquids. In this way, the analyzer 200 does not come into direct contact with the sample P and/or other fluids and/or reagents and can in particular be reused for another test without being disinfected and/or cleaned first.

It is however provided that the analyzer 200 is connected or coupled mechanically, electrically, thermally and/or fluidically and/or pneumatically to the cartridge 100.

In particular, the analyzer 200 is designed to have a mechanical effect, in particular for actuating the pump apparatus 112 and/or the valves 115, and/or to have a thermal effect, in particular for temperature-controlling the reaction cavity/cavities 109 and/or the intermediate temperature-control cavity 110 and/or the sensor apparatus 113.

In addition, the analyzer 200 can preferably be pneumatically connected to the car tridge 100, in particular in order to actuate individual apparatuses, and/or can be electrically connected to the cartridge 100, in particular in order to collect and/or transmit measured values, for example from the sensor apparatus 113 and/or sensor portions 116.

The analyzer 200 preferably comprises a pump drive 202, the pump drive 202 in particular being designed for mechanically actuating the pump apparatus 112. The analyzer 200 preferably comprises a connection apparatus 203 for in particular electrically and/or thermally connecting the cartridge 100 and/or the sensor arrange ment or sensor apparatus 113. As shown in Fig. 1 , the connection apparatus 203 preferably comprises a plurality of electrical contact elements 203A, the cartridge 100, in particular the sensor arrange ment or sensor apparatus 113, preferably being electrically connected or connecta ble to the analyzer 200 by the contact elements 203A. The analyzer 200 preferably comprises one or more actuators 205 for actuating the valves 115. Preferably, different (types or groups of) actuators 205A and 205B are provided which are assigned to the different (types or groups of) valves 115A and 115B for actuating each of said valves, respectively. Mostly preferred, the analyzer 200 comprises one or more actuators 205A for actuating the initially closed valves 115A and one or more actuators 205B for the normally open valves 115B.

The analyzer 200 preferably comprises one or more sensors 206. In particular, fluid sensors 206A are assigned to the sensor portions 116 and/or are designed or in tended to detect liquid fronts and/or flows of fluid in the fluid system 103.

Mostly preferred, the fluid sensors 206A are designed to measure or detect, in par ticular in a contact-free manner, for example optically and/or capacitively, a liquid front, flow of fluid and/or the presence, the speed, the mass flow rate/volume flow rate, the temperature and/or another value of a fluid in a channel and/or a cavity, in particular in a respectively assigned sensor portion 116, which is in particular formed by a planar and/or widened channel portion of the fluid system 103.

Alternatively or additionally, the analyzer 200 preferably comprises one or more (other or additional) sensors 206B for detecting the ambient temperature, internal temperature, atmospheric humidity, position and/or alignment, for example by means of a GPS sensor, and/or the orientation and/or inclination of the analyzer 200 and/or the cartridge 100.

The analyzer 200 preferably comprises a control apparatus 207, in particular com- prising an internal clock or time base for controlling the sequence of a test or assay and/or for collecting, evaluating and/or outputting or providing measured values in particular from the sensor apparatus 113, and/or from test results and/or other data or values.

The control apparatus 207 preferably controls or feedback controls the pump drive 202 and/or actuators 205, in particular taking into account or depending on the de sired test and/or measured values from the sensor apparatus 113 and/or sensors 206.

Optionally, the analyzer 200 comprises an input apparatus 208, such as a keyboard, a touch screen or the like, and/or a display apparatus 209, such as a screen.

The analyzer 200 preferably comprises at least one interface 210, for example for controlling, for communicating and/or for outputting measured data or test results and/or for linking to other devices, such as a printer, an external power supply or the like. The interface 210 might be embodied as a wired or wireless interface 210.

The analyzer 200 preferably comprises a power supply 211 for providing electrical power, preferably a battery or an accumulator, which is in particular integrated and/or externally connected or connectable.

Preferably, an integrated accumulator is provided as a power supply 211 and is (re)charged by an external charging device (not shown) via a connection 211 A and/or is interchangeable. The analyzer 200 is preferably portable or mobile. Preferably, the analyzer 200 weighs less than 25 kg or 20 kg, mostly preferred less than 15 kg or 10 kg, in partic ular less than 9 kg or 6 kg.

The analyzer 200 preferably comprises a housing 212, preferably wherein all the components and/or some or all of the apparatuses of the analyzer 200 are integrated in the housing 212 and/or arranged in the interior space 212A thereof.

Mostly preferred, the cartridge 100 can be inserted or slid into the housing 212, and/or can be received by the analyzer 200, through an opening 213 which can in particular be closed, such as a slot or the like. The analyzer 200 preferably comprises a pressurized gas supply 214 in order to provide a pressurized working medium, preferably gas, in particular air.

The pressurized gas supply 214 is preferably integrated in the analyzer 200 or the housing 212 and/or can be controlled or feedback controlled by means of the control apparatus 207.

Preferably, the pressurized gas supply 214 is electrically operated or can be oper ated by electrical power. In particular, the pressurized gas supply 214 can be sup- plied with electrical power by means of the power supply 211.

Fig. 4 shows the analyzer 200 in the open state/position, i.e. when the receptacle 201 is accessible and/or the opening 213 is formed. Here, the cartridge 100 has already been inserted into the analyzer 200, preferably through the opening 213 into the receptacle 201.

The analyzer 200 or housing 212 preferably comprises an access cover / housing part 212B that can be opened. Preferably, the analyzer 200, in particular its housing 212, can be opened by moving the access cover / housing part 212B relative to the housing 212, in particular a base 212C thereof, and/or such that the opening 213 is formed and/or the receptacle 201 is accessible, mostly preferred from the top.

Fig. 5 is an exploded view of the analyzer 200, showing its preferred construction. As already mentioned, the analyzer 200 preferably comprises a housing 212 that contains/encompasses the main, in particular all, (mechanical and/or electrical) parts/components of the analyzer 200.

The analyzer 200 preferably comprises a preferably mechanical closing/clamping system, the pressurized gas supply 214, at least one ventilation apparatus, at least one electronic unit and/or a support/cushion.

The ventilation apparatus is preferably adapted to ventilate/cool the analyzer 200 or housing 212, in particular its interior 212A. The top 212D of the housing 212 is preferably adapted to close/cover the analyzer 200, in particular its interior 212A, mostly preferred such that the analyzer 200, in particular the housing 212, is closed/protected/shielded to all directions. Preferably, the top 212D comprises or forms the opening 213, that is accessible by moving the access cover/housing part 212B relative to the housing 212, in particular its top 212D.

The housing 212, in particular its base 212C and top 212D, forms and/or limits the interior 212A, in particular laterally.

Preferably, the housing 212 and/or its interior 212A is at least essentially cuboid. However, other constructional solutions are possible as well, e.g. wherein the hous ing 212 and/or its interior 212A is at least essentially cylindrical.

In particular, a motor 202A of the pump drive 202 drives a pump head 202B of the pump drive 202.

The pump drive 202 and/or pump head 202B points towards the cartridge 100 and/or towards the intermediate unit 230, and therefore the pump head 202B can act on the pump apparatus 112 of the cartridge 100 in the desired manner in the test position. In particular, a fluid (gas or liquid) can be conveyed in the pump apparatus 112 and, thus, in the cartridge 100 by rotating the pump head 202B. The pumping is thus controlled by operating the pump drive 202 and/or pump motor 202A accordingly.

Preferably, a plurality of apparatuses of the analyzer 200, such as the actuators 205, and/or a plurality of apparatuses of the cartridge 100, such as the pump apparatus 112 and the sensor apparatus 113, can be supplied by the pressurized gas supply 214 and/or controlled and/or operated by the control apparatus 207 by activating corresponding valves and correspondingly supplying pressurized gas or pressurized air from the pressurized gas supply 214.

Following a test, the measurement results are read out electrically from the sensor apparatus 113 and processed either in the analyzer 200 or an external device (not shown). The analyzer 200 comprises preferably a clamping system 280 and/or two units 230, 231 for receiving, positioning and/or clamping, connecting and/or clamping the car tridge 100. In particular, a receiving unit 230 and a connecting unit 231 can be moved relatively to each other to position the cartridge 100 in between so that the cartridge 100 can be clamped, connected, held and/or positioned, in particular for performing the amplification and/or testing.

The system 10 or analyzer 200 comprises preferably an inductor 295 for inductively heating the at least one heating element 165.

Preferably, the inductor 295 is arranged at the clamping system 280 and/or at the receiving unit 230 and/or connection unit 231.

In the shown embodiment, the inductor 295 is preferably arranged at the connection unit 231 which is preferably unmoveable and/or provided with other electrical com ponents, contacts or the like.

The inductor 295 is preferably electrically operated to generate a varying magnetic flux acting on or at the associated heating element(s) 165 in the cavity/cavities 109, and/or cartridge 100 to induce electric currents in the heating element(s) 165 to in ductively heat the heating element(s) 165, in particular for the desired amplification reaction(s).

Fig. 6 shows a schematic view of the proposed heating element 165. Fig. 7 shows a schematic cross section of the heating element 165.

In particular, the heating element 165 has an extension in at least one dimension, preferably a length, width and/or high and/or diameter, greater than 10 pm, preferably greater than 100 pm, more preferably greater than 1 mm.

The heating element 165 can in particular be designed in such a way that it ensures a large surface area for the amplification of nucleic acids.

Preferably, the heating element 165, in particular a core 165A of the heating element 165, comprises or consists of an electroconductive material, preferably a magnetic or ferromagnetic material, in particular a nickel-iron alloy. This electroconductive ma terial can be inductively heated by the inductor 295. More preferably, the heating element 165 is ring-like, foil-like or wire-like.

In particular, the heating element 165 comprises an at least essentially areal or flat shape, preferably general or outer shape.

The heating element 165 is preferably non-spherical.

Fig. 6 shows that the heating element 165 is preferably wire-like or made of wire.

Preferably, the heating element 165 is or forms a helical ring or coil.

Preferably, the heating element 165 is a helically formed wire, in particular a sub stantially, preferably closed, circular and/or elliptical helically formed wire. In particu- lar, the heating element 165 is designed as a helically formed wire which is further provided as a closed loop or ring - as shown in Fig. 6. Such a design of the heating element 165 results in a large surface area which can be provided or functionalized with oligonucleotides 01 , 02, preferably primers, and/or other reagents. A high large outer surface of the heating element 165 is preferred for the amplifica tion, in particular for the polymerase chain reaction.

Especially preferably, the heating element 165 is designed as a single-turn or multi ple-turn coil and/or as a closed conductor loop, especially for improved induction interaction with the inductor 295.

Particularly preferably, the heating element 165 or wire can have an outer diameter of greater than 1 pm, preferably greater than 5 pm, more preferably greater than 10 pm or 100 pm and in particular between 10 pm and 20 pm or 50 pm.

Preferably, the outer diameter of the wire cross section of the heating element 165 can range between 5 and 40 pm or up to 300 pm, more preferably between 10 and 30 pm. Fig. 7 shows that the heating element 165 comprises preferably a core 165A and an outer coating 165B surrounding the core 165A. In particular, the core 165A can have an outer diameter between 1 and 50 pm, more preferably between 5 and 20 pm. The sheath thickness of the outer coating 165B can in particular range between 0.1 pm and 10 pm, preferably between 0.5 pm and 1 pm.

Particularly preferred, the material of the core 165A differs from the material of the coating 165B.

In particular, the core 165A enables the inductive heating of the heating element 165. Additionally or alternatively, the outer coating 165B can be heated inductively. Preferably, the core 165A comprises or consists of an electroconductive material, preferably a magnetic or ferromagnetic material, in particular a metal alloy, more preferably nickel-iron alloy.

Preferably, the coating 165B contains or consists of a material suited for the func- tionalization with the oligonucleotides 01 , 02, in particular of a precious metal, more preferably silver and/or gold.

Especially preferred, the heating element 165 comprises a core 165A with a nickel- iron alloy material and an outer sheathing of gold. In particular, the heating element 165 is designed as a sheathed wire.

Alternatively, the core 165A can contain or comprise a stainless steel, in particular a ferromagnetic stainless steel. Preferably, the core 165A has a high strength such that the wire can be preferably designed to be very thin.

In particular, a high outer surface compared to the volume ratio of the heating ele ment 165 is enabled.

The material of the outer coating 165B provides in particular the desired chemical properties.

As already mentioned, at least one heating element 165 is placed in at least one reaction cavity 109. This is shown schematically in Fig. 8 as a partial section of the system 10 or cartridge 100 or reaction cavity 109 with the heating element 165. Particularly preferred, the heating element 165 can be placed inside or inserted in the reaction cavity 109 before closing the cavity 109 - here by cover 102 - and or during assembly and/or before introducing the sample P into the reaction cavity 109. Figs. 2 and 8 show, that the heating element 165 is arranged preferably completely in the respective reaction cavity 109 to contact directly the sample P in order to heat it.

Inside the reaction cavity 109 the amplification of at least one analyte or nucleic acid sequence(s) takes place, in particular by a polymerase chain reaction (PCR).

The PCR or PCRs are carried out on the basis of protocols of temperature profiles that are essentially known to a person skilled in the art. In particular, the mixture of the sample volume located in the reaction cavity/cavities 109 is preferably cyclically heated and cooled. The heating of the sample P inside the reaction cavity 109 is achieved by the heating element 165.

In particular, only a part of the sample P received in the reaction cavity 109 is heated by the heating element 165, namely preferably the part close to the (surface of the) heating element 165.

Preferably, nucleic acid products and/or target nucleic acid sequences are produced as amplification product(s) in the reaction cavity/cavities 109. In particular, the amplification cycle consists of steps of denaturation, hybridization and elongation which are repeatedly passed, preferably in this sequence. In each passage of the cycle the number of nucleic acid molecules can be increased, so that in particular an exponential increase in the number of nucleic acids molecules can arise.

The nucleic acid sequence to be amplified can be preferably a single strand and can form, together with its complementary strand, a double strand. The nucleic acid to be amplified and the complementary strand can be a part of a larger nucleic acid. In particular in a PCR, a copy of the nucleic acid sequence to be amplified produced in one passage of the amplification cycle can form a template for formation of a com- plement (complementary strand) in a subsequent passage and a copy of the com plement produced can be a template for the formation of a nucleic acid sequence to be amplified in a subsequent passage of the cycle. Herewith, the explanation of the PCR process described in WO 2018/073435 A1 on page 4 to 5 is incorporated by reference.

As shown in Fig. 2, the heating element 165 is preferably loosely arranged in the reaction cavity 109.

Preferably, the heating element 165 is contact-less and/or wireless arranged in the reaction cavity 109.

The heating element 165 can in particular be produced separately from the other parts of the cartridge 100 and can more preferably be placed inside the reaction cavity 109 during assembly or closing of the cavity 109 and/or before introducing the sample P into the cartridge 100.

The heating element 165 has in particular not to be positioned in a predetermined manner inside the reaction cavity 109.

Alternatively, the heating element 165 can be fixed to the reaction cavity 109.

Mostly preferably, the heating element 165 comprises a main, in particular areal, extension or plane oriented at least essentially parallel to a main plane H of the car tridge 100 or a main plane of the inductor 295 or flat side or cover 102 of the cartridge 100 or reaction cavity 109. In particular, the main surface extension of the heating element 165 is at least substantially parallel to the flat side of the cartridge 100 and/or to the main plane and/or bottom plane of the reaction cavity 109. This can preferably improve the inductive interaction in order to heat inductively the heating element 165.

In particular, the heating element 165 is placed and/or inserted in such a way into the reaction cavity 109 and/or the heating element 165 is designed in such a way, that the inductor 295 which is provided for the induction of the heating element 165 and which is preferably arranged adjacent to the reaction cavity 109 can interact, in particular by inductively heating, with a large area and/or surface of the heating ele ment 165. Fig. 8 shows that the reaction cavity 109 comprises preferably an inlet 109D and an outlet 109E for fluid exchange. Preferably, the heating element 165 does not extend and/or protrude to or into the inlet 109D and/or outlet 109E.

In particular, for the amplification the inlet 109D and/or the outlet 109E or respective channels 114 are at least substantially liquid-tight sealed so that the sample P is enclosed in the reaction cavity 109.

Preferably, the heating element 165 is designed in such that the inlet 109D and/or outlet 109E are impassable for the heating element 165. Especially preferably, the inlet 109D and/or outlet 109E has a smaller diameter and/or cross section than the heating element 165, preferably by at least a factor of 2, 3, 4 or 5.

As shown in Fig. 8, the heating element 165 is designed in such that in at least one dimension the extension of the heating element 165 is larger than the cross section of the inlet 109D and/or the outlet 109E so that the heating element 165 cannot es cape from the reaction cavity 109 via the inlet 109D and/or outlet 109E.

Particularly preferably, the heating element 165 is functionalized and/or conjugated with one or more oligonucleotides 01 , 02 for the amplification of the at least one nucleic acid sequence, in particular for polymerase chain reaction. The functionali zation of the heating element 165 enables in particular that the surface area of the heating element 165 can provide the PCR reaction which can take place on and/or close to the outer surface of the heating element 165.

Particularly preferably, the heating element 165 can be functionalized with two or more different oligonucleotides 01 , 02 as schematically shown in Fig. 7, optionally for amplifying different analytes or nucleic acid sequences. Preferably, the PCR uses at least two different oligonucleotides 01 , 02, which are described as "primers": a forward primer and a reverse primer. The usage of the different primers for the PCR is described in WO 2018/073435 A1 on page 5 second and third paragraph which is herewith incorporated by reference.

At least one of the required primers and/or oligonucleotides 01 , 02 is fixed and/or functionalized on the heating element 165, in particular in order to allow the amplifi cation product to form and to facilitate a denaturation by local heating. Due to the functionalization of the heating element 165 a localization of steps of the amplification of nucleic acid(s), in particular hybridization, elongation and/or denaturation, as well as preferably also the generation of a signal to observe the process of PCR, is achieved in the direct vicinity of the heating element 165, the heating of the sample P can be limited to a fraction of the sample P volume received in the reaction cavity 109.

In particular, the oligonucleotides 01 , 02 are functionalized to the outer surface of the heating element 165.

Most preferably, the heating element 165 can be functionalized or bonded with a forward and reverse primer for the amplification. In a further preferred embodiment, the heating element 165 can be functionalized with a forward primer or a reverse primer. If the heating element 165 is only function alized with a forward primer, the reverse primer can be freely suspended in the sam ple P received in the reaction cavity 109 and vice versa. Preferably, different reagents, in the present case dry reagents S, particularly pref erably primers, in particular those required for the PCR or PCRs, in particular groups of different primers for different analytes in this case, are preferably added to the (premixed) sample P or the sample portions and/or are located in the intermediate cavity/cavities 106 arranged before or upstream of the respective reaction cavities 109 and/or in the reaction cavity / cavities 109, respectively.

In particular, the different primers and/or different oligonucleotides 01 , 02 differ in particular in terms of the hybridisation temperatures and/or the sequences of the am plification products generated by the respective primers.

Particularly preferably, marker primers are used in the sense already specified at the outset. Especially preferably, the heating element 165 can be functionalized and/or provided with the oligonucleotides 01 , 02 and/or other reagents for the application in such a way that a further or additional (dry) reagent, such as dry reagents S in cavity 106 - which would be suspended in the sample P before the sample P separately or is received in the reaction cavity 109 - can be avoided.

Mostly preferred, the dry molecules or reagents needed for the amplification are func tionalized to or provided by the heating element 165. By introducing the heating ele- ment 165 to the reaction cavity 109 preferably at least essentially all molecules or reagents needed for the amplification are provided.

Preferably, the heating element 165 is provided with an outer surface that allows the bonding of nucleic acids or primers. The primer can be bonded in particular via one or more thiol bond(s) on or to the heating element 165.

Particularly preferably, a gold or gold-plated surface of the outer coating 165B can be used in order to bind oligonucleotides 01 , 02, in particular primers, preferably via one or more thiol bond(s).

In particular, a streptavidin biotin bond can be used to bind a primer/oligonucleotides 01 , 02 to the heating element 165, in particular wherein the one or two partners (streptavidin or biotin) has been bonded (preferably beforehand) to the heating ele ment 165 and the primer/oligonucleotides 01 , 02 is modified with the other of the two partners and subsequently thereby bonded to the heating element 165.

Moreover, other modifications like amino or carboxy groups can also be used to bind primers/oligonucleotides 01 , 02 to the heating element 165, in particular to the outer coating 165B. For this purpose, the outer surface of the heating element 165 can in particular be modified with epoxy.

Particularly preferred, a bond is realized in such a way that the end of the primer/ol igonucleotides 01 , 02 (the so called 5'-end) is bonded to the heating element 165, wherein the other end of the primer/oligonucleotides 01 , 02 (the so called 3'-end) is free and can therefore be elongated during the amplification process, in particular during PCR by polymerase. Especially preferred, for the reason that the amplification of nucleic acid(s) occurs in the region close to the outer surface of the heating element 165 a heating of the whole sample in the reaction cavity 109 can be avoided. The temperature required for the PCR is preferably only achieved in a part of the sample P, in particular the part close to the heating element 165. Therefore, the en ergy required for the PCR or the amplification of nucleic acid(s) can be reduced ac cording to the invention. In particular, the heating element 165 is only heated for a short time, in particular by providing electromagnetic field provided by the inductor 295 for a short time, prefer ably in order to carry out the denaturation of the nucleic acids molecules in the sam ple P, while the majority of the sample P remains at a base temperature, in particular a temperature which is suitable for an elongation, preferably also a hybridization. This is preferably achieved by the duration of the heating through the heating element 165 being so short that the thermal field arising in the surrounding sample P can only spread a few micrometers and in this way creates a heating-up zone, which prefera bly comprises only a tiny fraction of the sample P. Preferably, the amount of heat brought can be so low that no substantial local heating of the sample P volume takes place.

The time needed for the amplification of nucleic acids can therefore be decreased. As stated in WO 2018/073435 A1 on page 11 , third paragraph - which is herewith incorporated by reference - the total process can only take a few milliseconds in total.

According to a further preferred aspect of the present invention, in the reaction cavity 109 several heating elements 165 are arranged.

Especially preferred, in one reaction cavity 109 at least two heating elements 165 can be introduced being functionalized with different oligonucleotides 01 , 02, in par ticular a forward and a reverse primer. Especially preferably, the plurality of heating elements 165 can differ from each other, in particular being functionalized with different oligonucleotides 01 , 02 and/or being of a different design. In particular, in at least one reaction cavity 109 at least two heating elements 165 can be arranged with a different functionalized outer surface and/or with a different design, preferably in order to accelerate the amplification of nucleic acid(s). Particularly preferably, in at least two different reaction cavities 109 different heating elements 165 can be inserted, in particular the different heating elements 165 are functionalized with different oligonucleotides 01 , 02 and/or are of a different design.

Preferably, the primer suspended in the sample P (forward and/or reverse primer) can be introduced to the sample P via dry / dried reagents S placed in the interme diate cavity 106 arranged before or upstream of the respective reaction cavities 109.

Particularly preferably, the reaction cavities 109 are full in succession with a specified volume of the "pretreated" sample P with respective sample portions via the interme- diate cavities 106 that are each arranged upstream of the respective reaction cavity 109. In the intermediate cavity 106 and/or in the reaction cavity 109 dry reagents S can be placed which can be solved in the sample volume.

In particular, a reverse and/or forward primer can be provided as a dry reagent S in the intermediate cavity 106 and/or in the reaction cavity 109. Particularly preferably, it is provided for a plurality of amplification reactions or PCRs to be carried out in parallel or independently from one another using different primers and/or primer pairs, such that a large number of (different) analytes A or target nucleic-acid se quences can be copied or amplified in parallel and subsequently analyzed.

In the embodiment shown in Fig. 2, the dry reagents or primers S are preferably contained in the intermediate cavities 106. However, other solutions are also possi ble, in particular those in which the dry reagents or primers are contained in the re action cavities 109- preferably together with the oligonucleotides 01 , 02 being func- tionalized on the outer surface of the heating element.

According to a preferred embodiment, the intermediate cavities 106 arranged before or upstream of the respective reaction cavities 109 each contain primers for amplify ing/copying one analyte, preferably two different analytes and more preferably three different analytes. However, it is also possible for four or more different analytes to be amplified/copied per reaction cavity 109 or sample portion. In particular, the heating element 165 is provided with one or more dried / dry sub stances or reagents S for amplification reaction. Fig. 7 shows that the heating ele ment 165 is provided with at least two different dry substances or reagents S1 , S2. Fig. 8 shows also the system 10 for amplification of at least one nucleic acid se quence of an in particular biological sample P, wherein the system 10 comprises at least one reaction cavity 109 for receiving the sample P, at least one inductively heatable heating element 165 and at least one inductor 295 for inductively heating of the heating element 165.

The inductor 295 is shown schematically in Fig. 8 by the dashed lines.

The heating element 165 is arranged completely in the reaction cavity 109 to contact directly the sample P in order to heat it.

In particular, at least or only part of the sample P is heatable by the heating element 165, in particular the part of the sample P close to the outer surface of the heating element 165. Further, Fig. 8 shows that the inductor 295 is arranged adjacent to the reaction cavity 109 for inductively heating the heating element 165.

In a preferred embodiment of the invention, the inductor 295 is arranged in the or engages into optional depression 101 E, as shown in Fig. 3.

The inductor 295 enables in particular a contactless heating of the heating element 165. Preferably, the inductor 295 is not directly connected to the heating element 165. In particular, the induction heating has numerous advantages over a conventional heating mechanism. It is a direct process. Preferably, the heat is thus generated in the heating element 165 itself and does not have to be introduced via its surfaces. In addition, the heating element 165 does not need to be contacted with further electri cal connection means.

The induction enables preferably a high process reliability and heating quality. The range in which the electromagnetic field acts in the material of the heating ele ment 165 - also known as the penetration depth - can be specifically influenced via the frequency. The frequency of the electromagnetic field can in particular range be tween 10 Hz and 1000 Hz, preferably between 20 Hz and 150 Hz. The frequency in particular allows a desired heating profile to be set precisely.

Especially preferred, the inductor 295 comprises a coil which is designed in such a way that the heating element 165 can be heated via the inductor 295. In particular, the system 10 comprises the cartridge 100 for receiving the sample P according to at least one of the embodiments described above. Further, the system 10 can comprise the analyzer 200 comprising the inductor 295 for testing the in par ticular biological sample P and inductively heating the heating element 165. Preferably, the analyzer 200 is designed to receive the cartridge 100 or to connect said cartridge 100 mechanically, electrically, thermally, fluidically and/or pneumati cally.

The cartridge 100 comprises the reaction cavity 109 and the heating element 165. The analyzer 200 further comprises the inductor 295, which can be connected to or arranged on the cartridge 100.

According to the proposed method for amplification at least or only part of the sample P is heated by at least one heating element 165 in the reaction cavity 109, wherein the heating element 165 is heated inductively by the inductor 295.

In particular, the oligonucleotides 01 , 02 can be bonded to the heating element 165 before the PCR is carried out or during the hybridisation step of the PCR. The heating step of the inventive method is preferably required for the denaturation step of the PCR.

Especially preferably, the heat supply by the means of the heating element 165 to the sample P or the part of the sample P close to the heating element 165 varies in such a way that the heat supply is in particular provided for the denaturation step but not for the hybridisation and/or elongation step of the PCR. Mostly preferred, the heated part of the sample P can be cooled using the unheated part of the sample P, which in particular surrounds the heated part of the sample P close to the heating element 165. The reachable heating and cooling of the sample P with regard to the PCR is de scribed in WO 2018/073435 A1 on page 33 to 36 which is herewith incorporated by reference.

Especially preferred, a cartridge 100 according to at least one of the embodiments described above and/or a system 10 according to least one of the embodiments de scribed above is/are used.

Preferably, during the nucleic-acid assay, a desired volume of the sample P that is mixed and/or pretreated in the mixing cavity 107 is preferably fed to one or more reaction cavities 109, particularly preferably via (respectively) one of the optional in termediate cavities 106 arranged before or upstream of the respective reaction cav ities 109. In particular, in the sample P that is fed to the reaction cavity 109 different reagents or primers, preferably dry reagents S, are added or dissolved. Particularly preferably, in particular during the nucleic-acid assay, the (premixed) sample P is split into several sample portions, preferably of equal size, and/or is divided between the several intermediate cavities 106 and/or reaction cavities 109, preferably evenly and/or in sample portions of equal size. Particularly preferably, the reaction cavities 109 are filled in succession with a spec ified volume of the (pretreated) sample P or with respective sample portions via the intermediate cavities 106 that are each arranged upstream of the respective reaction cavities 109. For example, the first reaction cavity 109 is filled with a specified volume of the pretreated sample P before the second reaction cavity 109 and/or the second reaction cavity 109 is filled therewith before the third reaction cavity 109.

Particularly preferably, the valves 115 that are assigned to the reaction cavities 109, and are in particular upstream and downstream, are sequentially opened, preferably such that the reaction cavities 109 can be individually or sequentially loaded with the sample P or the respective sample portions, and/or such that the sample P can be divided into a plurality of sample portions assigned to the reaction cavities 109. Preferably, the reaction cavities 109 are filled by (continuously) pumping using the pump apparatus 112, in particular until the sample P or the corresponding sample portion reaches the sensor portion 116 arranged directly downstream or thereafter, and/or until a flow of fluid is detected in the sensor portion 116 arranged directly downstream or thereafter. This ensures that the reaction cavities 109 are completely filled, and/or that the next method step, in particular the amplification, can only be initiated once the reaction cavities 109 have been completely filled.

As the reaction cavities 109 are being filled with the sample P or the corresponding sample portions, the fluid located in the reaction cavities 109, in particular the air located in the reaction cavities 109, is displaced and/or fed to a downstream cavity, for example the receiving cavity 104 or the mixing cavity 107. Preferably, the (pre treated) sample P is removed from the bottom of the mixing cavity 107 and at the same time the fluid, in particular the air, displaced by the sample P or sample portions is fed to the mixing cavity 107 at the top, in particular until the reaction cavities 109 are completely filled with the sample P or the respective sample portions.

Preferably, the sample portions are handled or conveyed individually, independently and/or separately from one another in the remainder of the method sequence. How- ever, other variants of the method are also possible in which the sample P is only temporarily divided into sample portions, and/or in which the sample portions are brought back together and are handled or conveyed together in the further method sequence. After carrying out the amplification reaction(s), corresponding fluid volumes, sample portions and/or amplification products are detected or measured. This can be done in the cavity 109 or cavities 109 or separately, here by feeding the one or more fluid portions, sample portions and/or amplification products preferably in succession to the (common or same) sensor arrangement, in particular to the (common or same) sensor apparatus 113 and/or to the (common or same) sensor compartment 118, in particular via a group-specific and/or separate intermediate cavity 106 and/or via the optional (common) intermediate temperature-control cavity 110.

Individual aspects and features of the present invention and individual method steps and/or method variants may be implemented independently from one another, but also in any desired combination and/or order. List of reference signs: system 200 analyzer

201 receptacle cartridge 202 pump drive A front 202A motor B back 202B pump head main body 203 connection apparatus E depression 203A contact element film/cover 205 (valve) actuator fluid system 205A (valve actuator) for 115A receiving cavity 205B (valve actuator) for 115B metering cavity 206 sensor intermediate cavity 206A fluid sensor mixing cavity 206B other sensor storage cavity 207 control apparatus reaction cavity 208 input apparatus D inlet 209 display apparatus E outlet 210 interface intermediate temperature- 211 power supply control cavity 211 A connection collection cavity 212 housing pump apparatus 212A interior space sensor arrangement 212B housing part/access coverE contact 212C base H central region 212D top channel 213 opening valve 214 pressurized gas supplyA initially closed valve 230 receiving unit B initially/normally open valve 231 connection unit sensor portion 280 clamping system connection 295 inductor closure element heating element F liquid reagent core H main plane of cartridge

coating L label

01 , 02 oligonucleotide

P sample

S, S1 , S2 (dry) reagent

Claims

Claims:

1. Cartridge (100) for amplification of at least one analyte, preferably a nucleic acid sequence, of an in particular biological sample (P), the cartridge (100) comprising a reaction cavity (109) for receiving the sample (P), characterized in that the cartridge (100) comprises at least one inductively heatable heating element (165) with an extension in at least one dimension greater than 0.1 mm, wherein the heating element (165) is arranged completely in the reaction cavity (109) to contact directly the sample (P) in order to heat it. 2. Cartridge according to claim 1 , characterized in that the heating element (165) is ring-like, foil-like or wire-like.

3. Cartridge according to claim 1 or 2, characterized in that the heating element (165) forms a closed conductor loop or coil.

4. Cartridge according to one of the preceding claims, characterized in that the heating element (165) comprises an areal or flat shape.

5. Cartridge according to one of the preceding claims, characterized in that the heating element (165) comprises a main extension or plane oriented at least essen tially parallel to a main plane (H) or flat side of the cartridge (100), a cover (102) thereof, or the reaction cavity (109).

6. Cartridge according to one of the preceding claims, characterized in that the heating element (165) is a helical ring or coil.

7. Cartridge according to one of the preceding claims, characterized in that the heating element (165) is non-spherical. 8. Cartridge according to one of the preceding claims, characterized in that the heating element (165) is loosely arranged in the reaction cavity (109). 9. Cartridge according to one of the preceding claims, characterized in that the heating element (165) is contact-less or wireless arranged in the reaction cavity (109). 10. Cartridge according to one of the preceding claims, characterized in that the reaction cavity (109) comprises an inlet (109D) and outlet (109E) for fluid exchange.

11. Cartridge according to claim 10, characterized in that the heating element (165) is designed such that the inlet (109D) and/or the outlet (109D) is impassable for the heating element (165).

12. Cartridge according to claim 10 or 11 , characterized in that the inlet (109D) and/or the outlet (109E) has a smaller diameter and/or cross section than the heating element (165).

13. Cartridge according to one of claims 10 to 12, characterized in that the cartridge (100) comprises a fluid system (103) fluidically connected or connectable with the reaction cavity (109) via the inlet (109D) and/or outlet (109E) for conveying the sam ple (P) to the reaction cavity (109).

14. Cartridge according to one of the preceding claims, characterized in that the heating element (165) is arranged in the reaction cavity (109) before conveying the sample (P) to the reaction cavity (109). 15. Cartridge according to one of the preceding claims, characterized in that only one heating element (165) is arranged within the reaction cavity (109).

16. Cartridge according to one of the preceding claims, characterized in that the heating element (165) has an extension in at least one dimension greater than 1 mm.

17. Cartridge according to one of the preceding claims, characterized in that the heating element (165) or a core (165A) thereof comprises or consists of an electro- conductive material, preferably a magnetic or ferromagnetic material and/ora nickel- iron alloy. 18. Cartridge according to one of the preceding claims, characterized in that the heating element (165) or a coating (165B) thereof is functionalized with primers or oligonucleotides (01 , 02) and/or other reagents (S1 , S2) for the amplification. 19. Cartridge according to one of the preceding claims, characterized in that the heating element (165) or a coating (165B) thereof is provided with one or more dried reagents (S1 , S2) for the amplification.

20. Heating element (165) for amplification of at least one nucleic acid sequence, wherein the heating element (165) or a core (165A) thereof comprises or consists of an electroconductive or ferromagnetic material and has an extension in at least one dimension greater than 0.1 mm, characterized in that the heating element (165) forms a closed conductor loop or coil and/or comprises an at least essentially areal or flat shape, and that the heating element (165) or a coating (165B) thereof is functionalized with pri- mers or oligonucleotides (01 , 02) and/or other reagents (S1 , S2) for the amplification and/or is provided with one or more dried reagents (S1 , S2) for the amplification.

21. Heating element according to claim 20, characterized in that the heating element (165) is ring-like.

22. Heating element according to claim 20 or 21 , characterized in that the heating element (165) is foil-like.

23. Heating element according to claim 20 or 21 , characterized in that the heating element (165) is wire-like.

24. Heating element according to one of claims 20 to 23, characterized in that the heating element (165) is helical. 25. Heating element according to claim 24, characterized in that the heating element

(165) is a helical coil or ring.

26. Heating element according to one of claims 20 to 25, characterized in that the heating element (165) is non-spherical. 27. Heating element according to one of claims 20 to 26, characterized in that the heating element (165) has an extension in at least one dimension greater than 1 mm. 28. Heating element according to one of claims 20 to 27, characterized in that the heating element (165) or a core (165A) thereof comprises or consists of an electro- conductive material, preferably a magnetic or ferromagnetic material.

29. Heating element according to one of claims 20 to 28, characterized in that the heating element (165) or a core (165A) thereof comprises or consists of a nickel-iron alloy.

30. System (10) for amplification of at least one analyte, preferably a nucleic acid sequence, of an in particular biological sample (P), the system (10) comprising: a reaction cavity (109) for receiving the sample (P); and at least one heating element (165); characterized in that the heating element (165) is inductively heatable and arranged completely in the reaction cavity (109) to contact directly the sample (P) in order to heat it, and that the system (10) comprises an inductor (295) for inductively heating the heating element (165).

31. System according to claim 30, characterized in that the system (10) comprises a cartridge (100) for receiving the sample (P) according to one of the claims 1 to 19 and an analyzer (200) comprising the inductor (295) for inductively heating the heat- ing element (165) in the cartridge (100).

32. System according to claim 30 or 31 , characterized in that the heating element (165) is constructed according to one of claims 20 to 29. 33. System according to one of claims 30 to 32, characterized in that the heating element (165) is helical, in particular a helical ring or coil. 34. Method for amplification of at least one analyte, preferably a nucleic acid se quence, of an in particular biological sample (P), preferably by polymerase chain reaction, wherein the sample (P) is conveyed to a reaction cavity (109), and wherein the sample (P) is heated in the reaction cavity (109) for the amplification, characterized in that at least or only part of the sample (P) is heated by inductively heating a heating element (165) arranged completely inside and/or loose in the reaction cavity (109).

35. Method according to claim 34, characterized in that a cartridge (100) according to one of claims 1 to 19 is used.

36. Method according to claim 34 or 35, characterized in that a heating element (165) according to one of claims 20 to 29 is used. 37. Method according to one of claims 34 to 36, characterized in that a system (10) according to one of claims 30 to 33 is used.

38. Method according to one of claims 34 to 37, characterized in that the heating element (165) is in particular loosely arranged in the reaction cavity (109) before the liquid sample (P) is conveyed to or entered into the reaction cavity (109).

39. Method according to one of claims 30 to 38, characterized in that the heating element (165) is functionalized with primers or oligonucleotides (01 , 02) and/or other reagents (S1 , S2) for the amplification.

40. Method according to claim 38 or 39, characterized in that the heating element (165) is provided with one or more dried reagents (S1 , S2) for the amplification which are soluted by the sample (P).