Sample carrier for use in a rotation-based method for amplifying DNA and / or for detecting nucleic acids

Abstract

The invention relates to a sample carrier (1) for use in a rotation-based method for amplifying DNA and / or for detecting nucleic acids, comprising a main part (2) and at least one amplification chamber (3) which is formed in the main part (3). The sample carrier is characterized in that the at least one amplification chamber (3) has a thickness (d) of less than 1 mm, in particular in a range between 10 µm and 0.9 mm, preferably 10 µm and 0.5 mm.

Description

[0001] Sample carrier for use in a rotation-based method for amplification of DNA and / or determination of nucleic acids

[0002] The invention relates to a sample carrier for use in a rotation-based method for amplifying DNA and / or determining nucleic acids. Furthermore, the invention relates to a system comprising such a sample carrier and a method for operating a rotation device. Furthermore, the invention relates to the use of such a sample carrier for amplifying DNA and / or determining nucleic acids.

[0003] Rotation-based methods are used, for example, in the medical field. They are typically used to determine nucleic acids, such as DNA (deoxyribonucleic acid) or RNA (ribonucleic acid). These methods are also used to amplify DNA.

[0004] The polymerase chain reaction (PCR) is typically used to amplify DNA. PCR is the standard method for the specific amplification of DNA and is essential for many modern biological and biomedical in vitro processes. Real-time PCR, in which the progressive amplification of the target DNA is continuously monitored by fluorescent probes, is particularly widespread and can be used, for example, for the specific identification of infectious pathogens from patient samples. The miniaturization of the PCR method using microfluidic techniques offers several advantages over conventional, macroscopic PCR in terms of cost, speed, transportability, and the potential for complete integration and automation of multiple sequential processes.

[0005] Essential to the PCR method is thermal cycling, in which the liquid temperature in the amplification chamber containing the relevant biological components is cyclically adjusted to the temperatures required for the biological processes. The speed at which this temperature change can be carried out has a direct impact on the overall effectiveness of the PCR and the duration of the entire cycle, with faster PCRs becoming increasingly important, for example, for point-of-care diagnostics.

[0006] For this reason, there are a variety of techniques specifically designed to accelerate temperature change. Examples include shunting microfluidic PCR, in which the liquid is moved back and forth between different heating zones, continuous-flow PCR, in which the liquid flows through different temperature zones, and methods that do not rely on contact heating, such as heating with lasers or microwave radiation. The most widespread and dominant method for commercial use is still temperature control by contact heating / cooling. The decisive factor for the speed of the temperature change is the volume of the liquid, which is directly proportional to the thickness of the chamber in relation to its contact surface with the contact heater.For thinner chambers, the temperature of the PCR fluid can be adjusted more quickly between the relevant temperatures than for thicker chambers, while the contact area to the heater is the same for both chambers. This makes it desirable to perform PCR in very thin chambers.

[0007] However, the smaller the size scales become, the greater the relative effects of surface tension and wetting. Therefore, as the chamber size decreases, it becomes increasingly difficult to remove air bubbles from a readout area of ​​the amplification chamber. Air bubbles can arise in various ways, for example, through repeated, cyclic heating to temperatures close to the boiling point of water, degassing of gases dissolved in the reaction volume can occur, or the dissolution of lyophilized samples can generate or promote bubbles. However, bubbles are a known interference factor for optical readout of DNA amplification and / or detection in the microliter range. Furthermore, flat amplification chambers are difficult to fill without bubbles. This requires special geometries, which complicate the manufacturing process of the sample carrier.The bubble formation that occurs in very thin chambers with small liquid volumes can have a significant impact on correct signal detection, especially when bubbles are located in the direct readout path of the optical readout device.

[0008] The object of the invention is therefore to provide a sample carrier by means of which the duration of the PCR process is reduced.

[0009] The object is achieved by a sample carrier for use in a rotation-based method for amplifying DNA and / or determining nucleic acids, comprising a base body and at least one amplification chamber formed in the base body, characterized in that the at least one amplification chamber has a thickness of less than 1 mm (millimeter), in particular in a range between 10 pm (micrometer) and 0.9 mm, preferably between 10 pm and 0.5 mm. Furthermore, an object of the invention is to provide a method by means of which the duration of the PCR method is reduced.

[0010] The object is achieved by a method for operating a rotation device with a carrier holder which carries at least one sample carrier according to the invention, wherein the rotation device is operated in a readout mode or a venting mode, wherein the speed of the rotation device is lower in the readout mode than in the venting mode.

[0011] The sample carrier according to the invention has the advantage of increasing the effectiveness of the overall PCR cycle by reducing the duration of the overall PCR cycle. This is possible because the selected thickness of the amplification chamber allows the temperature of the liquid in the amplification chamber to change quickly. In other words, the liquid in the amplification chamber can be heated or cooled to the target temperature very quickly. This is because thin amplification chambers result in small absolute spatial temperature differences and / or small temperature gradients within the liquid. The resulting more uniform temperature distribution allows for homogeneous reaction conditions in the amplification chamber, which is beneficial for shortening the duration of the overall PCR cycle.

[0012] The sample carrier according to the invention thus makes it possible to reduce the time required for the PCR process described above. At the same time, when the sample carrier is rotated by the rotation device, the amplification chamber can be easily read by the optical readout device without gas bubbles, especially air bubbles, interfering with the readout process. This will be explained in more detail below.

[0013] For the purposes of the application, the term "thickness" has two different meanings. According to a first meaning, "thickness" is understood to mean the extension of the amplification chamber in a direction that runs along the rotation axis of the sample carrier and / or is perpendicular to the base body. The thickness of the amplification chamber can vary along a radial and / or azimuthal direction. Therefore, as described in more detail below, several sections of the amplification chamber can exist that have different thicknesses along the rotation axis. Therefore, it is not excluded that a section of the amplification chamber has a thickness of at least 1 mm. "Radial" and "azimuthal" refer to the rotation axis of the sample carrier and / or the carrier holder.

[0014] According to a second meaning, the "thickness" may correspond to an "average thickness" which, as described in more detail below, is determined taking into account a liquid volume and a base area of ​​the amplification chamber.

[0015] The thickness according to the first meaning differs from the "thickness" according to the second meaning, i.e., the "average thickness," in the way it is determined, as described in more detail below. For example, the thickness of the amplification chamber according to the first meaning can be measured, while the liquid volume and a heated base area are also used to determine the average thickness.

[0016] The average thickness is the quotient between a liquid volume, in particular a heated or cooled one, within the amplification chamber divided by a heated or cooled base area of ​​the amplification chamber. In a design in which several heated or cooled base areas are present, the sum of the heated or cooled base areas is used to determine the average thickness. The heated or cooled base area can correspond to the base area of ​​the amplification chamber, which is wetted with liquid, in particular during operation of the rotating device, preferably during the PCR process. In a design with several heated or cooled base areas, the sum of the part of the base areas that is wetted with liquid is used to determine the average thickness.

[0017] The heated or cooled base surface allows the heat, in particular the significant portion, to be introduced into the amplification chamber or removed from the amplification chamber. The heated or cooled base surface of the amplification chamber can correspond to at least a portion of the floor area of ​​the amplification chamber. By taking the average thickness into account, the time required to perform the PCR process can be reduced. This is possible because the time required for the PCR process depends significantly on the heat input into the liquid. The average thickness takes this heat input into account.

[0018] "Readout operation" refers to an operation of the rotating device in which a readout of at least one amplification chamber takes place. "Readout" refers to a process in which, using the optical readout device, it is determined whether nucleic acids are present in the amplification chamber. A data processing device of the rotating device can determine whether nucleic acid is present in the amplification chamber based on the readout data. Alternatively or additionally, the data processing device can determine the replication of the DNA in the amplification chamber based on the data acquired during the readout process. For the purposes of the application, "rotational speed of the rotating device in readout operation" means that the rotating device is also rotated during readout operation. Thus, the readout operation takes place with a rotating rotating device, enabling real-time PCR.

[0019] The electrical or electronic data processing device can have at least one processor or can be a processor. The data processing device can be part of a circuit board. Furthermore, the data processing device can be arranged in an interior of the rotating device or outside the interior of the rotating device.

[0020] In venting mode, the sample carrier rotates at at least a specified speed. During venting mode, the readout device does not read the amplification chamber. The speed in venting mode is higher than in readout mode. Alternatively, the speed in venting mode can be the same as in readout mode. In venting mode, the speed of the carrier holder is sufficiently high to expel gas bubbles from the amplification chamber.

[0021] A rotation device can be operated in rotation mode. In rotation mode, the sample carrier is rotated at at least a predetermined speed. In rotation mode, the amplification chamber can be read by the readout device; alternatively, at particularly high speeds, the amplification chamber cannot be read by the readout device. In rotation mode, the added sample, in particular a biological sample, is processed by the reagents contained in the sample carrier. This allows a PCR cycle to be carried out in rotation mode. The speed in rotation mode can be higher than the speed in readout mode and / or in venting mode. This is necessary because a minimum speed is required to process the sample, and the speed of the detector limits the maximum speed during readout.However, it cannot be ruled out that the rotational speed in rotation mode corresponds to the rotational speed in readout mode or is lower than the rotational speed in readout mode. A rotation device comprising at least one sample carrier according to the invention and a rotation device having a rotatable carrier holder that holds the sample carrier is also particularly advantageous. The rotation device has the advantage that the interfering gas bubbles can be removed from the amplification chamber by rotating the sample carrier. This is possible because the forces generated on the air bubbles by the rotation cause the gas bubbles to no longer be located in the optical path of the readout device.

[0022] More specifically, the rotational forces enable a constant removal of gas bubbles from the readout section of the amplification chamber. In particular, the displacement of the gas bubbles is achieved by a higher rotational speed of the carrier holder in venting mode than in readout mode, which pushes the liquid in the amplification chamber radially outward, thus displacing the lighter gas bubbles radially inward. "Radially in" or "radially out" refers to the radial direction relative to the rotation axis of the sample carrier and / or the carrier holder.

[0023] The sample carrier according to the invention enables reliable real-time PCR for thin amplification chambers, particularly for amplification chambers with an average thickness in the range of 10 μm to 0.5 mm. Due to the thin design of the amplification chamber, rapid adjustment of the liquid in the amplification chamber to a target temperature is possible. This enables rapid thermal cycling and thus fast real-time PCR.

[0024] The polymerase chain reaction (PCR) is an artificial method for amplifying DNA. As described above, thermal cycling is essential for the PCR process. During thermal cycling, the liquid temperature in the amplification chamber containing the relevant biological components is cyclically adjusted to the temperatures required for the biological processes. The PCR process involves multiple PCR cycles, during which the aforementioned temperature adjustment process takes place. The large number of cycles is necessary to increase the number of DNA strands. Therefore, it is not uncommon for the PCR process to involve 30-50 PCR cycles.

[0025] In real-time PCR, the progressive amplification of the target DNA is determined, whereby the readout section of the sample carrier can be read during a PCR cycle.

[0026] The determination of nucleic acids can comprise the detection of nucleic acids and / or the quantification of nucleic acids. In this case, the nucleic acid can be determined in the entire amplification chamber, provided the optical readout device reads the entire amplification chamber. Alternatively, a readout section of the amplification chamber can be read, leaving the remaining section of the amplification chamber unreadable.

[0027] In one embodiment, a readout device for reading the amplification chamber can be provided. The part of the amplification chamber that is filled with liquid can be read out. The readout device can be suitable for determining nucleic acids, in particular DNA and / or RNA. The readout device can be configured to optically detect the amplification chamber. Thus, the readout device can comprise an objective lens and at least one optical element, such as at least one deflecting mirror and / or optical filter. The readout device can be a fluorescence detector. In this case, the readout device is configured to detect and / or process fluorescence signals emanating from the amplification chamber.For this purpose, fluorescent probes located in the amplification chamber can be read using the reading device and a conclusion can be drawn about the replication of the DNA located in the amplification chamber.

[0028] The at least one amplification chamber of the sample carrier can have sections with different thicknesses. "Thickness" refers to the first meaning, i.e., the actual extension of the amplification chamber along the rotation axis of the sample carrier in a region of the section. The sections therefore have different thicknesses in the radial and / or azimuthal direction of the sample carrier. In this embodiment, too, the average thickness of the amplification chamber corresponds to the quotient between the liquid volume and the heated or cooled base area, in particular the part of the heated or cooled base area wetted with liquid.

[0029] The at least one amplification chamber can have two sections with different thicknesses along the rotation axis of the sample carrier, with the deeper of the two sections being readable. An amplification chamber designed in this way has the advantage of ensuring good readout. Thus, the readout device cannot read the entire amplification chamber, but only a readout section of the amplification chamber. Preferably, in particular, only a readout section of the amplification chamber is read out, with the readout section being deeper than a remaining section of the amplification chamber. A deeper chamber increases the probability that more target material, such as DNA and / or RNA, is arranged in the readout section of the amplification chamber. With a higher amount of target material in the

[0030] A stronger signal can also be received by the readout device in the readout section.

[0031] The target material, in particular the nucleic acid, can be directly determined using the readout device. Alternatively, it is possible to determine a different material from which the target material, in particular the nucleic acid, can then be determined. This allows a by-product formed during a reaction to be identified, and from this, the amount of nucleic acid in the amplification chamber to be determined. With a fluorescent measurement method, the by-product fluoresces and can be determined by the optical readout device.

[0032] The readout section can be arranged radially between at least two residual sections of the amplification chamber relative to the rotation axis of the sample carrier. This means that there can be a first residual section that is radially closer to the rotation axis than the readout section. Furthermore, there can be a second residual section that is radially farther from the rotation axis than the readout section. Reading in the deeper readout section offers the advantage that more material can be read out compared to reading in the residual section. This improves the signal-to-noise ratio.

[0033] The readout section of the at least one amplification chamber can have a thickness, along the rotation axis of the sample carrier, of at least 1 mm. In particular, the readout section can have a thickness, along the rotation axis of the sample carrier, in a range between 1 mm and 2 mm. The remaining section of the amplification chamber can have a thickness, along the rotation axis of the sample carrier, of less than 1 mm. In particular, the amplification chamber can have a thickness, along the rotation axis of the sample carrier, in a range between 10 μm and 0.9 mm, preferably between 10 μm and 0.5 mm.

[0034] In particular, the sample carrier can have an amplification chamber with a readout section having an average thickness of less than 1 mm, in particular in a range between 10 pm and 0.9 mm, preferably between 10 pm and 0.5 mm. In such an embodiment, the thickness can vary in the radial and / or azimuthal direction.

[0035] Another advantage of an amplification chamber with sections of varying depth is that fluid can be pumped out of the amplification chamber through the deep amplification chamber section, i.e., the amplification chamber section with a thickness of at least 1 mm. The shallow amplification chamber section, i.e., the amplification chamber section with a thickness of less than 1 mm, has a high resistance, which counteracts rapid and reproducible pumping of fluid out of the amplification chamber.

[0036] The readout section can comprise at least 10% of the volume of the amplification chamber, in particular a range between 10-50% of the volume of the amplification chamber. The readout section includes the part of the amplification chamber that is filled with liquid. In particular, the readout section includes a base area, in particular the floor area, of the amplification chamber that is wetted with liquid. In contrast, a section of the amplification chamber that does not contain liquid cannot be read.

[0037] A readout section designed in this way enables accurate reading of the readout section and the detection of target material, particularly fluorophores, located there. At the same time, the readout section is not too large, allowing rapid temperature changes in the liquid in the amplification chamber. The rotation of the sample carrier results in thermal convection, which quickly and evenly distributes heat throughout the amplification chamber. Therefore, the deeper readout section does not have a significant adverse effect on the rapid and consistent temperature changes of the liquid in the amplification chamber.

[0038] In a special design, the thickness can correspond to an average thickness. The average thickness corresponds to the ratio of the liquid volume within the amplification chamber divided by the heated or cooled base area of ​​the amplification chamber. A particularly advantageous design is one in which the measurement is not based on the entire heated or cooled base area, but only on the part of the base area that is wetted with liquid.

[0039] Each amplification chamber can be assigned a prechamber in which the liquid collects before being introduced into the amplification chamber. The supply of liquid from the prechamber to the amplification chamber can occur when the prechamber is completely filled with liquid and / or can be controlled by the rotational speed of the rotating device. Since the dimensions of the prechamber are known, the portion of the amplification chamber's base area that is wetted with liquid is also known after the liquid has been introduced from the prechamber into the amplification chamber. By focusing on the average thickness, the duration of the overall PCR cycle is further reduced. This is because the average thickness is determined exclusively by considering the portion of the heated or cooled base area that is wetted with liquid.This allows the actual heat input into the liquid to be taken into account, thus shortening the PCR process time. This takes advantage of the fact that at the surface wetted with the liquid, heat transfer occurs directly into the liquid. At the unwetted surface, the heat input first occurs into the air and then from the air into the liquid. This heat input is much less efficient and therefore negligible.

[0040] The liquid-wetted base area of ​​the amplification chamber may correspond to the entire base area of ​​the amplification chamber, in particular the entire floor area of ​​the amplification chamber, through which heat is introduced into the amplification chamber or removed from the amplification chamber.

[0041] Alternatively, the liquid-wetted base area of ​​the amplification chamber can be smaller than the total base area of ​​the sample carrier, in particular the total floor area of ​​the amplification chamber, through which heat is introduced into or removed from the amplification chamber. In other words, designs are possible in which the liquid does not wet the entire floor area of ​​the amplification chamber. In these designs, the liquid-wetted base area, in particular the floor area, of the amplification chamber differs from the support surface of the sample carrier, which rests on the carrier holder and is introduced into or removed from the amplification chamber via heat.

[0042] The amplification chamber, in particular the main amplification chamber, can extend in the radial direction in a range between 1 mm and 10 mm, in particular 5 mm, and / or can extend in the azimuthal direction in a range between 1 mm and 6 mm, in particular 4 mm. An amplification chamber designed in this way has a sufficiently large volume so that a high quantity of target material is likely to be arranged in the amplification chamber. At the same time, the amplification chamber is not too large, so that a temperature change of the liquid in the amplification chamber can occur quickly. A pre-amplification chamber can be designed differently from the main amplification chamber. In particular, the pre-amplification chamber can extend 20 mm in the radial direction and / or 20 mm in the azimuthal direction.

[0043] The amplification chamber can be a main amplification chamber or a pre-amplification chamber. The sample carrier can have one or more main amplification chambers. Furthermore, the sample carrier can have one or more pre-amplification chambers. The sample carrier can also have at least one main amplification chamber and at least one pre-amplification chamber. The sample, particularly a biological sample, introduced into the sample carrier can first be processed in the pre-amplification chamber before being processed in the main amplification chamber.

[0044] By providing at least one pre-amplification chamber, the amount of target material, such as nucleic acid, that enters the main amplification chamber can be increased by a factor of 100 or more. This ensures that enough target material is present in the main amplification chamber for the target material to be detected by the readout device. This avoids the problem of no target material being present in the main amplification chamber. This problem arises because the main amplification chamber is flat.

[0045] Furthermore, providing at least one pre-amplification chamber has the advantage of achieving high robustness against inhibitors. In a PCR pre-amplification with 5-20 cycles, the quantities of DNA copies produced are typically not comparable to those produced in a one-step PCR with 30-50 cycles. As a result, biomolecules such as primers or polymerase are usually not limited, and requirements for their processivity and activity in reaction environments are less critical. Furthermore, the inhibition mechanisms typical of real-time PCR, which disrupt fluorescence generation, do not apply to pre-amplification. The disruption only becomes significant from the second amplification in the main amplification chamber onwards, where a dilution factor of typically 5-40 is present.

[0046] A sample carrier can be designed such that a pre-amplification of the nucleic acid is performed in the pre-amplification chamber before the nucleic acid is amplified in the main amplification chamber. The pre-amplification and main amplification take place after the sample carrier is inserted into the rotation device. In particular, the pre-amplification and main amplification occur due to the rotation of the sample carrier by the carrier holder.

[0047] The main amplification chamber is understood to be the chamber in which the DNA is amplified and / or in which the nucleic acid, in particular DNA and / or RNA, is determined. The readout device is designed and arranged such that it reads the main amplification chamber. At least one reagent can be arranged in an amplification chamber, in particular a main amplification chamber and / or a pre-amplification chamber. If there are multiple amplification chambers, each amplification chamber can contain at least one reagent. The reagents can differ from one another, allowing different processing of the liquid introduced into the amplification chamber.

[0048] In a particular embodiment, the main amplification chamber can have at least one section with a thickness, in particular an average thickness, of less than 1 mm, in particular in a range between 10 μm and 0.9 mm, preferably between 10 μm and 0.5 mm. If the embodiment has a pre-amplification chamber, the pre-amplification chamber can have a thickness, in particular an average thickness, of less than 1 mm, in particular in a range between 10 μm and 0.9 mm, preferably between 10 μm and 0.5 mm, or of at least 1 mm. It is advantageous for the pre-amplification chamber to have a thickness, in particular an average thickness, of less than 1 mm so that the amplification process can be carried out quickly.

[0049] In one embodiment, the base body can be disc-shaped and / or circular segment-shaped, and / or a microfluidic channel and / or chamber structure can be present in the base body. The amplification chamber is a component of the microfluidic channel and / or chamber structure. A "microfluidic channel structure" is understood to mean a channel structure whose dimensions in at least one spatial direction, in particular in the depth direction and / or tangential direction, can range between 30 and 700 pm (micrometers).

[0050] The rotatable carrier holder of the rotation device can be designed to accommodate one or more sample carriers. In the case that the sample carrier is designed in the shape of a circular segment, the rotatable carrier holder can accommodate at least two sample carriers. The sample carriers can be rotated simultaneously by the carrier holder. The sample carrier can be designed and configured such that at least one of the subsequent method steps can be realized using the sample carrier. For example, the sample carrier can have an access line into which the sample to be processed is introduced. In addition, thermal and / or chemical and / or mechanical lysis can be carried out. Furthermore, the sample can be mixed with a pre-amplification buffer. The mixture can be mixed with a first lyophilizate containing enzymes for an amplification reaction. A pre-amplification, e.g.PCR can be performed in 50-500 μl volumes with 5-20 cycles. A portion of the pre-amplifier can be measured and mixed with main amplification buffer and a lyophilisate containing enzymes, such as polymerase, for the main amplification. The mixture of main amplification buffer, lyophilisate, and pre-amplifier can be divided into one or more main amplification chambers.

[0051] The sample carrier can have a film element that is attached to the base body and / or forms a floor of the amplification chamber. The amplification chamber can thus be delimited by the film element and base body walls. In particular, the film element can be attached, in particular directly, to a base body side and / or attached to the base body in such a way that the base body is sealed. The film element can be elastic. In particular, the film element can have a smaller modulus of elasticity than the base body. The film can have a thickness in a range between 30 pm and 200 pm, in particular 90 pm. The film element can rest on the carrier holder, in particular directly. Heat can be introduced by the carrier holder through the film element into the amplification chamber in order to heat the liquid contained in the amplification chamber.Heat can be transferred almost exclusively conductively from the carrier holder to the film element. Heat can be transferred convectively from the film element to the amplification chamber. Alternatively, heat can be transferred convectively from the amplification chamber to the film element.

[0052] In one embodiment, the base body can have one, in particular a single, inlet for introducing liquid into the amplification chamber. The liquid can contain the nucleic acid. A gas bubble located in the amplification chamber can be expelled from the amplification chamber through the inlet when the rotating device is in rotation mode. This means that the gas bubble can be expelled from the amplification chamber through the same inlet through which the liquid was introduced into the amplification chamber. Thus, no gas bubble chambers need to be provided in the amplification chamber, which simplifies the design of the amplification chamber. Furthermore, there is no need for the amplification chamber to have a separate outlet because the gas bubble can be expelled from the amplification chamber through the inlet for introducing the liquid.The inlet can be arranged between the amplification chamber and the pre-chamber in the case of a flow direction into the amplification chamber.

[0053] In one embodiment, the readout device and / or the sample carrier can be configured such that the amplification chamber is read out during a readout operation of the rotating device. The readout device and the rotatable carrier holder can be positioned opposite each other with respect to the sample carrier. In particular, the side of the sample carrier facing away from the film element can face the readout device.

[0054] The rotation device can comprise a device for heating or cooling the amplification chamber. The device can cool or heat a carrier holder section. The device can be a Peltier element and / or a resistance heating wire and / or can be arranged to heat or cool the carrier holder section of the carrier holder. The device is used to adjust the temperature in the amplification chamber.

[0055] The sample carrier can be arranged on the carrier holder in such a way that the temperature change of the liquid occurs, in particular exclusively, through the carrier holder. In particular, the sample carrier can be arranged, preferably directly, on the carrier holder, so that the amplification chamber is heated or cooled by the carrier holder. In other words, heat is introduced into the amplification device only from one side of the sample carrier. This enables a simple design of the rotation device because it is not necessary to provide heating or cooling devices at different locations on the rotation device. Alternatively, multiple devices can be provided for heating or cooling the liquid present in the amplification chamber.

[0056] The rotation device can have a vacuum device for applying a negative pressure to the sample carrier, in particular the film element. Negative pressure is understood to be a pressure that is less than atmospheric pressure. By applying a negative pressure, the film element and the base body are held to the carrier holder. This means that when the negative pressure is applied, the carrier holder and the sample carrier cannot be moved relative to one another. A further advantage of applying the negative pressure is that the film element is tightened. The tightening is necessary because otherwise the film element would be wavy due to its elastic design, causing the volume of the amplification chamber to fluctuate and / or be undefined, which makes reading the amplification chamber difficult. There is a need to use the thinnest possible film element in order to achieve the fastest possible temperature change in the amplification chamber.The speed of the temperature change depends on the thickness of the film element.

[0057] In a particular embodiment, the data processing device can have a rotary drive for driving the carrier holder. The data processing device can be connected to the rotary drive via data technology. A data connection is understood to be an electrical connection by means of which data can be exchanged between components. The data processing device can control the rotary drive such that the rotary device is operated, in particular selectively, in a rotation mode, a venting mode, or a readout mode. For this purpose, a corresponding control command can be transmitted from the data processing device to the rotary drive.

[0058] During venting mode, the rotational speed of the carrier holder is sufficiently high that the forces acting on the gas bubbles and the liquid are sufficiently high that the gas bubbles are forced radially inward, i.e., move toward the rotation axis. The rotation axis is perpendicular to the base body. The gas bubbles can be expelled from the amplification chamber, particularly exclusively, via the inlet.

[0059] The rotational speed of the rotating device in readout mode can be lower than in venting mode. The data processing device can cause the venting operation to be performed before a readout operation. This offers the advantage of ensuring that a majority or all of the gas bubbles are removed from the amplification chamber before the readout device optically reads the amplification chamber in readout mode. For the purposes of the application, "cause" means that the data processing device is configured to exchange data with other components of the rotating device in order to realize a technical effect, such as adjusting a rotational speed of the rotating device and / or causing the readout device to read the amplification chamber. In particular, the data processing device can receive data from a component and / or transmit control commands to a component.

[0060] The data processing device can cause the rotation device to be operated in venting mode at the start of operation and then to operate the rotation device only in readout mode or rotation mode. The data processing device transmits corresponding control commands to the rotary drive to implement readout mode or rotation mode. This takes advantage of the fact that most gas bubbles are created when the liquid is poured into the sample carrier and that no or almost no gas bubbles are created during rotation mode. In particular, no gas bubbles are generated during readout mode because the temperature of the liquid in the amplification chamber is lower in readout mode than in rotation mode.

[0061] The data processing device can be configured such that the rotation device is operated selectively in venting mode, in rotation mode, or in readout mode.

[0062] The speed of the sample carrier in readout mode can be between 1 Hz and 20 Hz, particularly between 2 Hz and 5 Hz. In contrast, the speed in rotation mode can be between 2 Hz and 120 Hz. The readout time can be between 1 second and 6 seconds. The readout time is the time required to read the amplification chamber(s). In particular, the readout time can be 1 to 2 seconds if the sample carrier is rotated at a speed of 5 Hz. Alternatively, the readout time can be 4 to 6 seconds if the sample carrier is rotated at a speed of 2 Hz.

[0063] In venting mode, the speed can be between 16 Hz and 120 Hz. The rotation device can be operated in venting mode for a maximum period of 2 minutes, in particular a period of between 1 minute and 2 minutes. The venting mode can be carried out first after the rotation device has been switched on. In particular, the venting mode can be carried out for a predetermined period of time. After the venting mode, the rotation device can be operated in readout mode or in rotation mode for at least one cycle, in particular for all cycles. The speed of the carrier holder in readout mode and / or rotation mode can be lower than in venting mode. In a special embodiment, the amplification chamber can be alternately heated and cooled, in particular during a PCR process.In particular, the temperature of the liquid in the amplification chamber can be cyclically changed between at least two temperatures. A cycle comprises the plurality of heating or cooling processes in which the liquid is brought to the different temperatures. Multiple cycles can be performed, each involving a change in the liquid temperature to at least two temperatures. The cycles can last a predetermined period of time and / or can be repeated at predetermined intervals. The individual cycles can differ from one another in their duration and / or the temperatures to be achieved during the cycles.

[0064] The data processing device can cause a readout operation to be performed during a PCR process. In particular, the data processing device can cause a readout operation to be performed during a PCR cycle of a PCR process and / or in a PCT cycle. Furthermore, the data processing device can specify when a readout operation should be performed in the PCR process. Thus, it is possible for a readout operation to be performed in every PCR cycle, every other PCR cycle, or in predetermined PCR cycles.

[0065] The recorded readout signal, especially the fluorescence signal, corresponds to a sigmoidal curve in the case of a positive signal. The shape of the curve can be used to determine the amount of DNA or RNA in the sample. Of particular relevance is the number of cycles after which the signal rises above a baseline value (Ct value). An early rise corresponds to a low Ct value with a high amount of analyte, while a late rise corresponds to a high Ct value with correspondingly little analyte.

[0066] A stable signal is essential for determining the Ct value. Gas bubbles, such as air bubbles, can cause significant jumps in the signal level and thus distort the Ct value. Therefore, for real-time PCR, it is essential that the readout area of ​​the amplification chamber is free of bubbles during the readout operation. Therefore, a venting operation can be performed during the PCR process before and / or after a readout operation, especially after each readout operation.

[0067] The data processing device can also cause the sample carrier to rotate during a PCR process. This ensures thorough mixing of the liquid in the amplification chamber. The thermal conductivity of the liquid in the amplification chamber, such as water, is not very high. Convection is therefore important for removing heat from the heated base surface. Rotating the sample carrier can assist this removal of heat from the base surface. This is because the density of warm liquid is typically and / or within a relevant temperature range lower than the density of cold liquid. During rotation, centrifugal forces create an artificial gravitational field in the amplification chamber. The liquid moves toward the center of rotation on the heated side and away from the center of rotation on the unheated side.During cooling, the process is equivalent, but reversed. Diffusion, Coriolis, and Euler forces result in faster mixing in the amplification chamber. This mixing ensures rapid heating and cooling even with one-sided heat input. Rapid mixing also has positive effects on the PCR reaction itself, because the reaction components are also continuously mixed. This reduces the time required for the PCR process.

[0068] Furthermore, rotating the sample carrier during the PCR process offers the following advantage. As already described above, DNA is typically amplified during a PCR process. However, many applications also require the detection of RNA. For this purpose, RNA is converted into DNA using a process called reverse transcription. Reverse transcriptase is used in this process. This enzyme is not usually temperature-stable and is inactivated at temperatures above 60°C. However, effective degassing of the liquid in the amplification chamber requires a temperature above 60°C. Degassing must therefore take place after reverse transcription and thus within the PCR process, especially between two PCR cycles.

[0069] The data processing device can cause the readout operation to be performed while the sample carrier is rotated. In other words, a readout operation is possible during the PCR process. This is advantageous because, with real-time PCR, a readout operation is often performed in every PCR cycle or every other PCR cycle. The data processing device can cause the device to heat or cool the amplification chamber while the sample carrier is rotated. Both of these functions are possible due to the above-described design of the sample carrier and the rotation device. As a result, real-time PCR can be performed due to the design of the sample carrier and the rotation device. The sample carrier can be connected to the rotation device in a rotationally fixed manner.Heat can be introduced into the amplification chamber by means of the heating or cooling device, or heat can be removed from the amplification chamber by means of the heating or cooling device, through the same sample carrier section, in particular the film element section. This enables a simply constructed sample carrier and / or a simply constructed rotation device.

[0070] It is particularly advantageous if a sample carrier according to the invention is used for the amplification of DNA and / or determination of nucleic acids.

[0071] The figures show the subject matter of the invention schematically, with identical or equivalent elements generally being provided with the same reference numerals. Here:

[0072] Fig. 1 shows a representation of a base body with amplification chambers of a sample carrier according to the invention.

[0073] Fig. 2 is an exploded view of a sample carrier according to the invention.

[0074] Fig. 3 is a plan view of a rotatable carrier holder of a rotation device.

[0075] Fig. 4 a side view of the carrier holder.

[0076] Fig. 5 is a plan view of the carrier holder on which the base body of the sample carrier according to the invention is mounted.

[0077] Fig. 6 is a plan view of the carrier holder on which the sample carrier according to the invention is mounted.

[0078] Fig. 7 is a side sectional view of a portion of the sample carrier according to a first embodiment with an amplification chamber.

[0079] Fig. 8 is a side sectional view of a portion of the sample carrier according to a second embodiment with an amplification chamber.

[0080] Fig. 9 is a side sectional view of a portion of the sample carrier according to a third embodiment with an amplification chamber.

[0081] Fig. 10a a sectional view from above of a base body section in a state in which the carrier holder does not rotate.

[0082] Fig. 10b is a top sectional view of a base body portion showing a state in which the carrier holder rotates.

[0083] Fig. 10c shows a top sectional view of the base body section shown in Fig. 10a at a later time than Fig. 10b. Fig. 11 shows a schematic representation of the rotation device.

[0084] Fig. 12 is a schematic representation of a part of the base body shown in Figure 2.

[0085] Fig. 1 shows a simplified representation of a base body 2 of a sample carrier 1 shown in Figure 2, which is used in a rotation-based method for amplifying DNA and / or determining, in particular for detecting and / or quantifying, nucleic acids. In the base body 2 shown in Figure 1, a plurality of channels and / or chambers formed in the base body 2 have been omitted compared to the base body 2 shown in Figure 2. Thus, in Figure 1, only a plurality of amplification chambers 3 are shown as examples, which are formed in the base body 3. In particular, two pre-amplification chambers 3b and five main amplification chambers 3a are shown. However, sample carriers 1 are conceivable which have a different number of pre-amplification chambers 3b and main amplification chambers 3a. The base body 2 is disc-shaped and circular segment-shaped.

[0086] The amplification chamber 3, in particular the main amplification chamber 3a, has a thickness, in particular an average thickness, of less than 1 mm (millimeter), in particular in a range between 10 μm (micrometer) and 0.9 mm, preferably between 10 μm and 0.5 mm. The main amplification chambers 3a and the pre-amplification chambers 3b are fluidically connected to each other. The amplification chambers 3 are part of a microfluidic channel and chamber structure shown in Figure 2.

[0087] Fig. 2 shows an exploded view of a sample carrier 1 according to the invention. The channel and chamber structure 8, which is formed in the base body 2, has a plurality of channels and chambers, in particular several main amplification chambers 3a and several pre-amplification chambers 3b, which are fluidically connected to one another. In the state shown in Figure 2, one side of the channel and chamber structure 8 is open.

[0088] The sample carrier 1 has a film element 6, by means of which the open side of the channel and chamber structure is concealed. The film element 6 acts as the base for the open channels and chambers in the base body 2 when the sample carrier 1 is arranged on the carrier holder 10 shown in Fig. 3. The film element 6 is attached directly to one side of the base body. The film element 6 is thus heat-sealed to the base body 2. As explained in more detail below, heat is introduced into or removed from the channel and chamber structure via the film element 6. On its chord side, the base body 2 has an access section 17 of an access line 32 shown in Figure 5, which protrudes radially from the base body 2 and is fluidically connected to the channel and chamber structure. A sample, in particular a biological sample, can be introduced into the access line and from there processed in individual chambers of the sample carrier.The access section 17 can be reversibly closed by means of a capsule 31 to allow the introduction of the sample and subsequent reclosure. The access section 17 and the access line 32 are designed such that a swab (not shown in the figures) containing the sample to be processed can be inserted into the access line.

[0089] In an initial state, i.e., in a state in which no sample has yet been introduced into the access port 10, the sample carrier 1 contains several pre-stored liquid reagents in various chambers. In addition, primers are stored in the pre-amplification chambers 3b. Further primers and probes, in the form of polynucleotides or oligonucleotides, are stored in front of the main amplification chambers 3a. The primer pairs in the pre-amplification chambers 3b are identical or different, depending on the specific objective of the analysis method and / or the specific sequence. Thus, the primers in the main amplification chambers 3a are identical in pairs to the primers in the pre-amplification chambers 3b, or, for example, are intended for a "nested PCR" known in the art and thus designed differently.

[0090] Lyophilisates containing, for example, enzymes, polymerase, dNTPs (deoxynucleoside triphosphates), salts, and / or other upstream reagents (e.g., PCR additives, nuclease inhibitors, cofactors of the enzymes involved, etc.) are stored in additional chambers. The access line can contain a lysis agent and process control agents, such as spores, fungi, phages, or artificially produced targets. A lysis chamber connected to the access line contains a lysis pellet, a magnet, and a grinding medium. The latter can be glass and / or zirconia particles, for example.

[0091] The sample body 1 also has a cover body 18, which is attached to the edge of the base body 1 facing away from the film element 6. With the exception of the edge, the cover body 18 does not lie directly against the base body 2, but rather there is an air layer between the cover body 18 and the base body 2. This can be seen in Figures 7 to 9. The cover body 18 can be fixed to the base body 2 by means of locking hooks in corresponding recesses 19 of the base body 2. The cover body 18 has a readout window 20. The readout window 20 is transparent and arranged such that the contents of the main amplification chambers 3a can be read out through the readout window 20. In addition, the cover body 18 has a recess 21, which serves to accommodate the access line attached to the base body 2.

[0092] The base body 2 has a plurality of openings 22, which serve for the precise alignment and positioning of the sample carrier on a carrier holder 10 of the rotation device 9, shown in Figure 3. Positioning pins 23 of the carrier holder 10 engage in these openings 22 for positioning and fixing in a rotation plane parallel to the surface of the carrier holder 10.

[0093] Fig. 3 shows a plan view of a rotatable carrier holder 10 and Fig. 4 shows a side view of the carrier holder 10. The carrier holder 10 is circular and serves to rotate the sample carrier 1 about a rotation axis R. The rotation axis R is perpendicular to the disc-shaped base body 2. The carrier holder 10 is designed such that it can carry two base bodies 2 shown in Figures 1 and 2.

[0094] The carrier holder 10 has a plurality of heating or cooling sections 30. A device 12 for heating or cooling the sample carrier 1 is arranged below the heating or cooling section 30, as viewed along the rotation axis R, as shown in Figures 6 and 7. The devices 12 can each be a Peltier element or resistance heating plates. The heating or cooling sections 30 are arranged offset from one another and can be designed as plates that cover the device 12. Furthermore, the carrier holder 10 has a carrier base 32. The heating or cooling sections 30 and devices 12 are arranged in recesses in the carrier base 32.

[0095] In addition, the carrier holder 10 has seals 24 that completely enclose one or more of the heating sections 30. The seals 24 each contain a plurality of holes (not shown), which are offset from one another along the seals D24. To fix the sample carrier 1 on the carrier holder 10, a vacuum can be applied to the holes using a vacuum device 13 shown in Figure 10. In the fixed state, the sample carrier 1, in particular the film element 6, rests against the seals 24. Figure 5 shows a plan view of the carrier holder 10, on which the base body 2 of the sample carrier 1 according to the invention is mounted. A swab containing the sample to be processed is arranged in the access line 32. The base body 2 rests on the carrier holder 10 and is connected to the carrier holder 10 in a rotationally fixed manner. As can be seen from Figure 5, the base body 2 covers only half of the carrier holder 10.Another base body 2, not shown in the figures, can be arranged on the remaining section of the carrier holder 10. In the embodiment shown in Figure 5, the sample carrier 1 is shown without the cover body 18. In contrast, in Figure 6, the sample carrier 1 is shown with the cover body 18.

[0096] Fig. 7 shows a side sectional view of an amplification chamber 3 according to a first embodiment. In particular, Figure 7 shows a side sectional view of a section of a base body 2 in which the amplification chamber 3 is arranged and of a part of the carrier holder 10, in particular the heating section 30 and the device 12. In the present exemplary embodiment, the amplification chamber 3 is the main amplification chamber 3a. In the example shown, the main amplification chamber 3a is only partially filled with liquid 33. An interface 35 between an air section and a liquid section in the amplification chamber 3 is shown in Figure 7. In an exemplary embodiment not shown, the amplification chamber 3 can additionally or alternatively be the pre-amplification chamber 3b.

[0097] Figure 7 shows that the film element 6 rests on the carrier holder 10. The carrier holder 10 has the device 12 for heating or cooling the sample carrier 1, which is arranged below the heating section 30. The device 12 can bring the liquid in the amplification chamber 3 to different temperatures.

[0098] The amplification chamber 3 has a thickness d along the rotation axis R of the sample carrier 1. The amplification chamber 3 is delimited by the film element 6 and by the walls of the base body 2. The rotation device 9 also has a readout device 11, which is arranged above the amplification chamber 3 along the rotation axis R. The readout device 11 can optically read the amplification chamber 3 and thus determine nucleic acids, in particular DNA, in the amplification chamber 3. A data processing device 14, which is connected to the readout device 11, can determine how the DNA has replicated in the amplification chamber 3 based on the received data. The carrier holder 10 has a temperature sensor 26. The temperature sensor 26 measures the temperature of the heating section 30.Since the film element 6 is thin, the temperature of the liquid within the amplification chamber 3 is almost the same temperature as the heating or cooling section 30.

[0099] Figure 7 shows a base area 29 of the amplification chamber 3, by means of which heat is introduced from the film element 6 into the amplification chamber 3. The base area of ​​the amplification chamber 3 corresponds to a floor area of ​​the amplification chamber 3 and is heated or cooled by the device 12. In the embodiment shown in Figure 7, the entire base area 29 is heated or cooled by the device 12. However, alternative embodiments (not shown) are conceivable in which only a portion of the base area 29 or the floor area of ​​the amplification chamber 3 is heated or cooled by the device 12.

[0100] In Figure 7, the liquid 33 wets only a portion of the base area 34 of the amplification chamber 29. An average thickness of the amplification chamber 3 corresponds to the quotient of the volume of the liquid 33 located in the amplification chamber 3 divided by the heated or cooled base area 34, i.e., the portion of the base area of ​​the amplification chamber 3 that is wetted with liquid 33. It is advantageous that the average thickness of the amplification chamber 3 is less than 1 mm, in particular between 10 pm and 0.9 mm.

[0101] Fig. 8 shows a side sectional view of an amplification chamber 3 according to a second embodiment. The second embodiment differs from the embodiment shown in Figure 7 in the design of the amplification chamber 3. The amplification chamber 3 shown in Figure 8 has two sections spaced apart from one another in the radial direction, i.e., in the direction Z, with different thicknesses d along the rotation axis R of the sample carrier 1. Thus, the amplification chamber 3 has a readout section 4, which is read by the readout device 11 during the readout operation of the rotation device 9. In addition, the amplification chamber 3 has a residual section 5, which is not read by the readout device 11. The readout section 4 is thicker than the residual section 5. The residual section 5, viewed in the radial direction, is arranged closer to a rotation axis (not shown in Figure 8) than the readout section 4.

[0102] The readout section 5 can be thicker than the remaining section 4 of the amplification chamber 3. The

[0103] Readout section 5 can have a thickness along the rotation axis R of the sample carrier 1 of at least 1 mm. To obtain the advantages of the invention, the average thickness of the amplification chamber 3 should be less than 1 mm, ideally between 10 pm and 0.9 mm. The readout section 5 is shown at the edge of the amplification chamber 4, but in some embodiments it can be located at any location within the amplification chamber 3. In the embodiment shown in Figure 8, the average thickness corresponds to the quotient of the liquid volume in the readout section 5 and the remaining section 4 divided by the portion of the heated or cooled base surface 34 that is wetted with liquid.

[0104] Fig. 9 shows a side sectional view of a section of the sample carrier according to a third embodiment with an amplification chamber 3. The third embodiment differs from the second embodiment shown in Fig. 8 in the design of the amplification chamber 3. Thus, the remaining section 5 is transferred into the readout section 4 by means of a bevel 36.

[0105] Fig. 10a shows a sectional view from above of a section of the base body 2 in which the amplification chamber 3 is formed, in a state in which the carrier holder 10 is not rotating. The amplification chamber 3 can be designed like an amplification chamber 3 shown in Figures 7-9. The carrier holder 10 is not shown in Figure 10a. In the illustrated state, gas bubbles 27 are arranged in the amplification chamber 3. Furthermore, liquid is arranged in the amplification chamber 3, whereby the amplification chamber 3 is only partially filled with liquid. In particular, the amplification chamber 3 is filled with liquid to 90% of its volume. However, since the base body 2 is not rotated, the liquid is distributed over the entire base area of ​​the amplification chamber 3.

[0106] Fig. 10b shows a sectional view from above of the base body section shown in Fig. 10a in a state in which the carrier holder 10 (not shown) is rotating. In particular, the rotation device 9 is operated in a venting mode. In the venting mode, the carrier holder 10 rotates about the rotation axis R and the carrier holder 10 is operated at a speed at which the forces acting on the liquid and the gas bubbles cause the gas bubbles to be displaced radially inward. The direction of force Z acting on the liquid and the gas bubbles is shown in Figure 10b. The gas bubbles can be guided out of the amplification chamber 3 through an inlet 28. The inlet 28 also serves to introduce liquid into the amplification chamber 3. Fig. 10c shows a sectional view from above of the base body section shown in Fig. 10a in a later state than Fig. 10b. As can be seen from Fig.As can be seen in Figure 10c, most of the gas bubbles have already been removed from the amplification chamber 3.

[0107] Fig. 11 shows a schematic representation of the rotation device 9. The rotation device 9 has a rotary drive 16 for driving the carrier holder 10. The rotary drive 16 is coupled to the carrier holder 10 and / or has a drive motor (not shown). The sample carrier 1 is arranged on the carrier holder 10. The readout device 11 is arranged above the sample carrier 1, as viewed along the rotation axis R.

[0108] The rotation device 9 also includes the data processing device 14. The data processing device 14 is connected for data purposes to the readout device 11, a vacuum device 13, and the rotary drive 16. The data processing device 14 can transmit control commands to the rotary drive 16 so that the rotation device 9 is operated in rotation mode or in readout mode.

[0109] The vacuum device 13 can be a pump that creates a vacuum in the bores to fix the sample carrier 1 to the carrier holder 10. The data connection is shown in dashed lines in Figure 11. The rotation device 9 has a housing 15 that encloses an interior space. The above-mentioned components are arranged within the housing 15.

[0110] Fig. 12 shows a schematic representation of a part of the base body 2 shown in Figure 2. In particular, Fig. 12 shows a schematic representation of part A of the base body 2 circled in Figure 2. Figure 12 shows three amplification chambers 3. The amplification chambers 3 can be designed identically to one of the amplification chambers 3 shown in Figures 7-9.

[0111] Each of the three amplification chambers 3 is assigned a pre-chamber 37. The pre-chamber 37 is fluidically connected to the inlet 28 of the amplification chamber 3. Furthermore, the pre-chambers 37 are fluidically connected to one another by means of a channel 38. The pre-chambers 37 are fluidically connected to the access line 17 of the sample carrier 1 by means of the channel 38. During rotation of the rotating device, liquid enters the pre-chambers 37 due to centrifugal force. The pre-chambers 37 are filled with liquid one after the other in chronological order. This means that first one pre-chamber 37 is completely filled with liquid and then the pre-chamber 37 located downstream in terms of flow is filled with liquid. The speed of the rotating device is selected such that the liquid in the pre-chambers 37 does not flow into the amplification chambers 3.This process is carried out until all prechambers 37 are filled with liquid.

[0112] The base body 2 has an overflow (not shown in the figures) through which excess fluid can be drained. This ensures that each amplification chamber 3 is filled exactly the same.

[0113] Subsequently, the speed of the rotating device and thus the speed of the sample carrier 1 are increased. By increasing the speed of the rotating device and thus the speed of the sample carrier 1, the liquid located in the prechambers 37 can flow into the respectively assigned amplification chamber 3. Since the volume of liquid located in the prechamber 37 is known, the portion of the base area 34 of the amplification chamber 3 that is wetted with liquid in the respectively assigned amplification chamber 3 is also known after the liquid flows into the amplification chamber 3.

[0114] List of reference symbols

[0115] 1 sample carrier

[0116] 2 basic bodies

[0117] 3 Amplification chamber

[0118] 3a Main amplification chamber

[0119] 3b Pre-amplification chamber

[0120] 4 Reading section

[0121] 5 Remaining section

[0122] 6 slide element

[0123] 8 Channel and chamber structure

[0124] 9 Rotation device

[0125] 10 carrier holders

[0126] 11 Reading device

[0127] 12 Device for heating or cooling

[0128] 13 Vacuum device

[0129] 14 Data processing device

[0130] 15 housings

[0131] 16 rotary drive

[0132] 17 Access section

[0133] 18 cover bodies

[0134] 19 Recess

[0135] 20 reading windows

[0136] 21 Recess

[0137] 22 Breakthrough

[0138] 23 Position pin

[0139] 24 Seal

[0140] 26 Temperature sensor

[0141] 27 gas bubbles

[0142] 28 Entrance

[0143] 29 Sample carrier section

[0144] 30 heating sections

[0145] 31 capsules

[0146] 32 support floor

[0147] 33 Liquid

[0148] 34 part of the base area wetted with liquid

[0149] 35 Interface

[0150] 36 slopes

[0151] 37 Antechamber

[0152] R rotation axis

[0153] Z force direction

Claims

Patent claims 1 . Sample carrier (1) for use in a rotation-based method for amplifying DNA and / or determining nucleic acids, with a base body (2) and at least one amplification chamber (3) formed in the base body (3), characterized in that the at least one amplification chamber (3) has a thickness of less than 1 mm, in particular in a range between 10 pm and 0.9 mm, preferably between 10 pm and 0.5 mm.

2. Sample carrier (1) according to claim 1, characterized in that a. the at least one amplification chamber (3) has sections with different thicknesses, in particular along a rotation axis (R) of the sample carrier (1), or that b. the at least one amplification chamber (3) has two sections with different thicknesses, in particular along a rotation axis (R) of the sample carrier (1), wherein the deeper of the two sections can be read out by a readout device (11).

3. Sample carrier (1) according to claim 1 or 2, characterized in that a. a readout section (4) of the at least one amplification chamber (3) has a thickness (d), in particular along a rotation axis (R) of the sample carrier (1), of at least 1 mm, in particular in a range between 1 mm and 2 mm, and / or a remaining section of the amplification chamber (3) has a thickness (d), in particular along a rotation axis (R) of the sample carrier (1), of less than 1 mm, in particular in a range between 10 pm and 0.9 mm, preferably between 10 pm and 0.5 mm, and / or that b. the thickness is an average thickness of the amplification chamber (3), wherein the average thickness corresponds to a quotient between a liquid volume within the amplification chamber (3) divided by a heated or cooled base area of the amplification chamber (3), in particular corresponds to a part of the heated or cooled base area (34) which is wetted with liquid.

4. Sample carrier (1) according to claim 3, characterized in that the readout section (4) has at least 10% of the volume of the amplification chamber (3), in particular a range between 10-50% of the volume of the amplification chamber (3).

5. Sample carrier according to one of claims 1 to 4, characterized in that the amplification chamber, in particular the main amplification chamber, extends in the radial direction in a range between 1 mm and 10 mm, in particular 5 mm, and / or extends in the azimuthal direction in a range between 1 mm and 6 mm, in particular 4 mm.

6. Sample carrier (1) according to one of claims 1 to 5, characterized in that the amplification chamber (3) is a main amplification chamber (3a) or a pre-amplification chamber (3b).

7. Sample carrier (1) according to one of claims 1 to 6, characterized in that a. the base body (2) is disc-shaped and / or circular segment-shaped and / or that b. a microfluidic channel and / or chamber structure is present in the base body (2).

8. Sample carrier (1) according to one of claims 1 to 7, characterized in that the sample carrier (1) has a film element (6) which is attached to the base body (2) and / or forms a bottom of the amplification chamber (3).

9. Sample carrier according to one of claims 1 to 8, characterized in that the base body (2) has an inlet (28) for admitting liquid into the amplification chamber (3), wherein the base body (2) is designed such that a gas bubble (27) located in the amplification chamber (3) can be discharged from the amplification chamber (3) through the inlet (28).

10. Rotation device (9) with at least one sample carrier (1) according to one of claims 1 to 9 and a rotatable carrier holder (10) which holds the sample carrier (1).

11. Rotation device (9) according to claim 10, characterized in that the rotation device (9) has a read-out device (11) for reading the amplification chamber (3), in particular the read-out section (4).

12. Rotation device (9) according to claim 11, characterized in that the reading device (11) and / or the sample carrier (1) is designed such that the amplification chamber (3) can be read out in a reading operation of the rotation device (9).

13. Rotation device (9) according to one of claims 10 to 12, characterized in that the rotation device (9) has a device (12) for heating or cooling the amplification chamber (3) or has a device (12) for heating or cooling the amplification chamber (3) which heats or cools a carrier holder section.

14. Rotation device (9) according to one of claims 10 to 13, characterized in that the rotation device (9) has a vacuum device (13) for applying a vacuum to the sample carrier (1), in particular the film element (6).

15. Rotation device (9) according to one of claims 10 to 14, characterized in that the rotation device (9) has a data processing device (14) which is connected for data purposes to the reading device (11) and / or a rotary drive (16) for the carrier holder (10).

16. Rotation device (9) according to claim 15, characterized in that the data processing device (14) causes the rotation device (9) to be operated in a read-out mode or a rotation mode or a venting mode, wherein the rotational speed of the carrier holder (10) is lower in the read-out mode than in the venting mode.

17. Rotation device (9) according to claim 15 or 16, characterized in that the data processing device (14) causes a venting operation to be carried out before a reading operation.

18. Rotation device (9) according to one of claims 10 to 17, characterized in that a. the rotational speed of the carrier holder (10) in a readout mode is between 1 Hz and 6 Hz, in particular 2 Hz to 5 Hz, and / or that b. the rotational speed of the carrier holder (10) in a venting mode is between 16 Hz and 120 Hz.

19. Rotation device (9) according to one of claims 10 to 18, characterized in that a readout time is between 1 second and 6 seconds.

20. Rotation device (9) according to one of claims 15 to 19, characterized in that a. the data processing device (14) causes a readout operation to be carried out during a PCR method or that the data processing device (14) causes a readout operation to be carried out during a PCR cycle of a PCR method and / or that b. the data processing device (14) causes the sample carrier (1) to be rotated during a PCR method, in particular during a PCR cycle.

21. Rotation device (9) according to one of claims 15 to 20, characterized in that a. the data processing device (14) causes the readout operation to be carried out while the sample carrier (1) is rotated and / or that b. the data processing device (14) causes the device (12) to heat or cool the amplification chamber (3) while the sample carrier (1) is rotated.

22. Rotation device (9) according to one of claims 15 to 21, characterized in that the sample carrier (1) is connected to the carrier holder (10) in a rotationally fixed manner, wherein heat is introduced into the amplification chamber (3) by means of the device (12) for heating or cooling or heat is removed from the amplification chamber (3) by means of the device (12) for heating or cooling through the same sample carrier section (29), in particular film element section.

23. Method for operating a rotation device (9), in particular according to one of claims 10 to 22, with a carrier holder (10) which carries at least one sample carrier (1) according to one of claims 1 to 9, wherein the rotation device (9) is operated in a read-out mode or a venting mode, wherein the speed of the rotation device (9) is lower in the read-out mode than in the venting mode.

24. Method according to claim 23, characterized in that a rotation operation of the Rotation device is carried out before the reading operation.

25. Method according to claim 23 or 24, characterized in that a. the rotational speed of the sample carrier (1) in readout mode is between 1 Hz and 6 Hz and / or that b. the rotational speed of the sample carrier (1) in rotation mode is between 16 Hz and 120 Hz.

26. Method according to one of claims 23 to 25, characterized in that a. the amplification chamber (3) is alternately heated and cooled and / or that b. the temperature of the amplification chamber (3) is cyclically changed between at least two temperatures. TI. Method according to one of claims 23 to 26, characterized in that a pre-amplification of the nucleic acid is carried out in a pre-amplification chamber (3b) before the nucleic acid is amplified in the main amplification chamber (3a).

28. Use of a sample carrier (1) according to one of claims 1 to 9 for the reproduction of DNA and / or determination of nucleic acids.

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