System for lamp amplification of DNA and / or RNA

Abstract

The invention relates to a system (400) for carrying out a LAMP amplification and for detecting the presence of DNA and / or RNA in a sample, the system consisting of the elements of a sample preparation device (200), a reaction vessel (100), and optionally a heating device (300). The system (400) allows a rapid detection of the presence of DNA and / or RNA in a sample, for example in order to detect the presence of a virus in the sample, using simple equipment.

Description

[0001] 40099 / 4361 / PTWO November 7, 2025

[0002] 1

[0003] System for LAMP amplification of DNA and / or RNA

[0004] Field of invention

[0005] The invention relates to a system for LAMP amplification of DNA and / or RNA, as well as elements of said system comprising a sample preparation device, a reaction vessel, and a heating device. Furthermore, a method for LAMP analysis is described, with which the presence of a specific DNA and / or RNA in a sample can be detected using the system presented here.

[0006] Background of the invention

[0007] For the detection of pathogenic material, such as viruses or bacteria, the so-called LAMP method (loop-terminated isothermal amplification), as described, for example, in Notomi et al., "Loop-mediated isothermal amplification of DNA", Nucleic Acids Research, 2000, Vol. 28, No. 12, No. E63, is known as a variant of isothermal DNA amplification. This method is successfully used, for example, to detect viral infections with the SARS-CoV-2 virus.

[0008] In the LAMP method, the target sequence to be detected, which can be DNA or RNA, is isothermally amplified in a sample at a constant temperature, usually between 60°C and 85°C, using at least one primer and one polymerase. Typically, four different primers are used to amplify six different regions of the target gene, which increases specificity. The additional use of so-called loop primers can further accelerate the reaction. In most cases, isothermal heating of the sample for 15 to 30 minutes is sufficient.

[0009] After completion of the isothermal heating, the amplified DNA or RNA can be detected using optical methods, for example, photometric methods or by the naked eye by exploiting a color change upon reaction of the DNA or RNA with another reagent (for example, complementary gold-bound ss-DNA).

[0010] US Patent 10,968,493 B1 describes a method for processing large numbers of samples in parallel. Such methods are helpful in the event of an epidemic, but they are not suitable for the early detection of viruses or bacteria (see 40099 / 4361 / PTWO, November 7, 2025).

[0011] 2. Generally, these methods are not helpful in a non-epidemic situation because they require complex equipment and necessitate the collection of numerous samples for parallel processing. As a result, the time between sample collection from the patient and availability of the test result is statistically comparatively long.

[0012] It would therefore be desirable to improve upon these disadvantages.

[0013] Summary of the invention

[0014] Based on this, the present invention aims to at least improve the disadvantages known from the prior art and, in particular, to simplify LAMP analysis using equipment and to shorten the time between taking a sample from the patient and obtaining the test result.

[0015] The task is solved by a sample preparation device for preparing a sample for performing a LAMP analysis to detect the presence of DNA and / or RNA in the sample, comprising

[0016] • a sample receiving chamber, preferably comprising a lysis buffer;

[0017] • a purification unit; and

[0018] • an eluate reservoir wherein the purification unit is part of a flask, wherein the sample receiving chamber is detachably connectable to the flask, so that when the sample receiving chamber is connected to the flask a pressure can be built up in the flask, by which a lysate, preferably formed in the sample receiving chamber by reaction of the sample with the lysis buffer, can be conveyed through the purification unit into the eluate reservoir, forming an eluate.

[0019] In this document, LAMP analysis refers to the analysis of whether a sample contains detectable DNA (deoxyribonucleic acid) or RNA (ribonucleic acid). This analysis is performed using the LAMP method. In the LAMP method, the target sequence, which can be DNA or RNA, is isothermally amplified (multiplied) in a sample at a constant temperature, typically between 60°C and 85°C, using at least one primer and one polymerase. Four different primers are generally used to amplify six different regions of the target gene, which increases specificity. The additional use of loop primers can further accelerate the reaction. The detection of the presence or absence of the target DNA or RNA is carried out according to Regulation (EU) No 40099 / 4361 / PTWO of November 7, 2025.

[0020] 3

[0021] Optical amplification, preferably by a color change reaction with a reagent specific to the DNA or RNA to be detected.

[0022] Preferably, the sample receiving chamber includes a corresponding lysis buffer through which the sample is lysed to form a lysate. Alternatively, it is also preferred that, instead of providing a lysis buffer within the sample receiving chamber, the lysis is carried out outside the chamber and the resulting lysate is introduced into the chamber. The sample receiving chamber is detachably connected to the piston and, for filling the sample or lysate into the chamber, is detached from the piston and then reconnected to the piston under pressure.

[0023] The flask contains the purification unit. This unit serves to purify the lysate, which is preferably generated in the sample receiving chamber. Impurities, such as other cell components that arise as unwanted byproducts during lysis, are retained in the purification unit, for example, by size exclusion processes.

[0024] The sample preparation device is preferably part of a system for performing LAMP analysis, which includes at least the reaction vessel described in this document and, optionally, the heating device also described in this document. However, it is also preferred to use the sample preparation device for LAMP analysis without the reaction vessel described in this document.

[0025] The sample preparation device includes the sample receiving chamber, into which a sample, obtained, for example, via a cotton ball or spatula, and which may include saliva, blood, stool, urine, tissue, and aqueous fluids containing RNA or DNA, can be scraped during lysis in the sample receiving chamber. This sample may contain the patient's own DNA and / or RNA, as well as DNA and / or RNA from other organisms, such as viruses. LAMP analysis is typically aimed at detecting the presence of specific DNA and / or RNA; for example, a virus may be identified based on its RNA in a patient's sample to prove that the patient is infected with the virus. 40099 / 4361 ZPTWO November 7, 2025

[0026] 4

[0027] For dispensing the sample into the sample receiving chamber, this chamber is preferably detachable from the rest of the sample preparation device or has a removable lid. The sample receiving chamber preferably includes a lysis buffer that breaks down the sample components after they are added to the chamber. The sample receiving chamber can be connected to a transfer vessel via a bayonet fitting or multiple bayonet fittings, which, once the connection is established, prevent unintentional disconnection. This prevents the release of the sample and any organism it may contain, thus avoiding the potential infection of other individuals.The sample preparation device is designed such that by connecting the sample receiving chamber to the transfer vessel or by placing the lid on the sample receiving chamber, a pressure is built up in the sample receiving chamber which transports the substance through the sample receiving chamber and the purification unit into the eluate reservoir.

[0028] After appropriate lysis in the sample receiving chamber or addition of the lysate, the lysate is transferred to the transfer vessel by the generated pressure. The purification unit, preferably consisting of a filtration column and a filter, forms a piston, preferably together with the transfer vessel. After being unlocked (preventing unintentional movement), the piston can be displaced against the housing of the sample preparation device, for example, by rotating it relative to the housing. This displacement causes plastic or elastic deformation of the eluate reservoir, building up pressure within the reservoir, which then allows the eluate to be conveyed, for example, into a reaction vessel.

[0029] Preferably, the piston is displaceable relative to the eluate reservoir in one direction of actuation. This makes it easy to seal the eluate reservoir by designing the piston with a cylinder that, when the piston is moved, is pushed over the outside of the eluate reservoir, sealing it airtight. The eluate reservoir preferably has one or more openings, perforations, or pores on its outer circumference, through which air displaced when the eluate is forced into the reservoir and when the piston is moved relative to it, can escape. Once the piston has reached a second position, the cylinder covers the outer circumference of the eluate reservoir, thus sealing the openings, perforations, or pores of the reservoir airtight.At this point, a defined portion, and in particular the entire sample, has been forced through the purification unit and is now present as eluate in the eluate reservoir. 40099 / 4361 / PTWO 7 November 2025.

[0030] 5

[0031] To transfer the eluate into a reaction vessel, particularly a reaction vessel as described below, the sample preparation device is positioned above the reaction vessel. Preferably, the sample preparation device has a positioning collar that forms a lower end of the device below the eluate reservoir. For the purposes of this document, "top" and "bottom" are to be understood as meaning that the sample preparation device has a first end at which the sample receiving chamber is formed. The direction towards this first end is referred to as "top," and the direction towards a second end of the sample preparation device opposite the first end is referred to as "bottom." The direction of actuation thus points from the first end to the second end of the sample preparation device.

[0032] Preferably, the positioning collar at the lower end of the sample preparation device is designed to interact with a positioning or centering aid of an access port of a reaction vessel, thereby ensuring optimal positioning of the sample preparation device relative to the reaction vessel. The sample preparation device preferably has a hollow cannula which is moved downwards, preferably against the action of a compression spring, by pressure on the piston of the sample preparation device in an actuation direction, until the hollow cannula protrudes above a lower edge of the positioning collar and can thus penetrate into the reaction vessel positioned below.

[0033] In the reaction vessel described below, the hollow cannula pierces a self-sealing first membrane of the vessel's access port. The hollow cannula is positioned precisely over the center of this self-sealing first membrane by the interaction of the positioning collar and centering aid. Simultaneously, pressure on the piston, which further displaces the piston, cylinder, and eluate reservoir downwards, forces the eluate reservoir against a stop in the reaction vessel, causing it to deform elastically and / or plastically. The resulting increased pressure within the eluate reservoir forces the eluate through the hollow cannula into the reaction vessel.

[0034] After the pressure on the piston is released, the compression spring causes a return movement, which moves the piston and eluate reservoir back into the second position. This ensures that the hollow cannula no longer protrudes beyond the lower edge of the positioning collar; the tip of the hollow cannula lies within the positioning collar. This prevents injury to a user of the sample preparation device and facilitates simple operation. 40099 / 4361 / PTWO 7 November 2025

[0035] 6

[0036] Disposal of the used sample preparation device should be possible without risk of injury.

[0037] The purification unit, preferably formed by a filtration column and filter, is adapted in its physical and chemical configuration to the type of analysis to be performed, and in particular to the DNA or RNA to be detected and / or the type of sample and / or lysate and its typical composition. In the preferably designed filtration column, corresponding chemical reactions and physical filtration processes take place, ensuring that the eluate contains the reactants required for the LAMP analysis in the reaction chamber, while other components of the sample and / or lysate that might inhibit or impede the corresponding reactions are filtered out. This can be achieved, for example, through size exclusion effects. The purification unit is preferably interchangeable to allow for easy adaptation of the sample preparation device to the specific DNA and / or RNA to be detected.

[0038] The sample preparation device according to the present invention allows for simple and rapid sample preparation for performing a LAMP analysis for detectable DNA and / or RNA. It is simple in design and can be used by personnel without medical training after a short familiarization period.

[0039] Preferably, a release unit is provided which allows the piston to be moved in the actuation direction towards the eluate reservoir when the release unit has been moved into an unlocked position. It is particularly preferred that the release unit can be moved into the unlocked position by a rotational movement of the piston.

[0040] Preferably, the release unit has at least one moving element that can move in a guide extending circumferentially to the piston. Preferably, the moving element is connected to the housing of the sample preparation device by a spring, which presses the spring radially inwards. Preferably, the housing has detents by which the moving element can be locked into a locking recess upon reaching it.

[0041] The release unit enables easy operation of the sample preparation device; the piston can only be moved in the direction of actuation by activating the release unit. By sealing the eluate reservoir externally with a cylinder of the piston, the eluate reservoir can be filled with 40099 / 4361 / PTWO 7 November 2025

[0042] 7. a porous wall or similar, which allows the displacement of air in the eluate reservoir by the injected eluate and at the same time allows this porous wall to be easily closed or covered after completion of the transfer of the eluate into the eluate reservoir.

[0043] The purification unit preferably comprises a filtration column and a filter. The filter preferably comprises a microporous plastic layer, for example made of high-density polyethylene (HDPE), with a pore size of several tenths of micrometers. The filtration column is preferably made of a silica gel-based material and achieves the necessary filtration, for example, via appropriately adapted size selection mechanisms.

[0044] Preferably, the purification unit is interchangeable. This allows the sample preparation device to be adapted to different types of DNA and / or RNA to be detected and / or to different sample types. For example, different filtration and purification processes are required for a stool sample than for a saliva or blood sample.

[0045] Preferably, a customized sample preparation device is available for a specific DNA / RNA to be detected, in which the lysis buffer and / or the purification unit are adapted to the type of sample (e.g., stool sample, saliva, blood, etc.) and / or the DNA / RNA to be detected. This is preferred regardless of whether a purification unit is interchangeable.

[0046] The object underlying the invention is also achieved by a reaction vessel for preparing a sample for LAMP analysis to detect the presence of DNA and / or RNA in the sample, comprising an access port for adding the sample, which is present as an eluate, into a fluid channel system connected to the access port. This fluid channel system connects the access port to at least two reaction chambers, each reaction chamber containing a lyophilized reaction solution specific to the DNA or RNA to be detected. At least the reaction chambers are sealed at an upper surface of the reaction vessel by a second membrane that is permeable to air but not to the eluate. The fluid channel system is preferably designed such that no intersecting channels for transporting different reagents are present.It is preferred that the access port be connected to a main channel, from which a secondary channel branches off, forming a fluid connection to a reaction chamber. 40099 / 4361 / PTWO 7 November 2025.

[0047] 8

[0048] The reaction vessel has at least two reaction chambers. This allows for different approaches. For example, it is preferably possible to always use one reaction chamber for a positive or negative control to verify the functionality of the test reaction. Alternatively or additionally, multiple reaction chambers allow for the detection of different pathogens, thus enabling on-site differential diagnosis of various pathogens. Preferably, each reaction chamber has a volume of no more than 30 mm³. 3 [cubic millimeters] on.

[0049] The reaction vessel is preferably plate-shaped. This means that the reaction vessel has a top surface and a bottom surface opposite the top surface, the area of ​​each of which is at least five times larger than the area of ​​the side surfaces or edges connecting the top surface and the bottom surface. Preferably, the top surface and bottom surface are aligned parallel to each other, at least in some areas.

[0050] The reaction solution comprises, on the one hand, the substances necessary for the LAMP reactions, such as primers, and on the other hand, preferably a color reagent which, in the presence of the DNA or RNA to be detected, causes a color change that is visible to the naked eye.

[0051] The reaction vessel is preferably made of plastic and preferably has a base body that forms the fluid channel system and the reaction chambers. This base body is preferably covered on the top side with the second membrane and preferably bonded to it by a material-bonded connection, in particular by gluing, welding, or injection molding, so that the second membrane preferably delimits the fluid channel system and / or the reaction chambers at least partially. The base body is preferably closed on the bottom by a base, which is preferably bonded to the base body by a material-bonded connection, in particular by gluing, welding, or injection molding. The base body is preferably manufactured by injection molding. The reaction vessel preferably has a thickness of less than 3 mm, preferably 2 mm or less, and most preferably less than 1.5 mm.This results in a low thermal mass of the reaction vessel, enabling simple and rapid heating, which is necessary for performing the LAMP analysis in the reaction chambers. The small volume of the reaction chambers, preferably a maximum of 30 mm³, is also advantageous. 3 [Cubic millimeters] simplifies heating.

[0052] The reaction vessel according to the present invention preferably interacts with the sample preparation device according to the present invention. (40099 / 4361 / PTWO 7 November 2025)

[0053] 9. The second membrane allows an eluate to be forced into the fluid channel system via the access port. The eluate flows through the fluid channel system into the reaction chambers, displacing the air present in the fluid channel system and the reaction chambers, which escapes through the second membrane.

[0054] Preferably, the access port is closed by a first, self-sealing membrane. This allows the first membrane to be pierced with a hollow cannula, for example, the sample preparation device as described above, to introduce the eluate under pressure, with the pressure being maintained when the hollow cannula is withdrawn from the self-sealing first membrane, causing it to close again. The first membrane is preferably made of a silicone material, more preferably a poly(organo)siloxane material.

[0055] Preferably, the reaction chambers are conically shaped. This reduces the volume of the reaction chamber and thus the volume of the sample to be analyzed. The reaction chambers preferably have a volume of approximately 20 to 50 pl [microliters]. With such volumes, LAMP analyses are reliably possible, and the volume of eluate and reaction solution that needs to be heated is comparatively small. This enables rapid amplification of the DNA / RNA to be detected. At the same time, the volume is large enough to ensure sufficient amplification of the DNA and / or RNA to produce a sufficiently large color change with the color reagent. This facilitates optical detection.

[0056] Preferably, the reaction vessel has a transparent bottom on a lower side opposite the top. This bottom is preferably made of a transparent plastic. The bottom is preferably bonded to a base body of the reaction vessel, preferably by welding, gluing, or injection molding. The transparent bottom allows visual verification of whether the color change has occurred due to the reaction of the DNA and / or RNA to be detected with the color reagent. For this purpose, after the LAMP analysis is complete, the reaction vessel is held with the bottom facing an observer, and the presence of the color change is then checked. To improve readability, the second membrane is preferably opaque and particularly preferably white. This further facilitates the visual verification of the color change, as the optical contrast is well-defined, especially with a white second membrane. 40099 / 4361 / PTWO November 7, 2025

[0057] 10

[0058] Particularly advantageous in this context is that the base of each reaction chamber has a viewing cone that tapers towards the top. This viewing cone further reduces the volume of the reaction chambers. Furthermore, when the bottom of the reaction vessel is turned upwards, any air remaining in the reaction chambers collects as bubbles at the tapered edges of the viewing cone, allowing an unobstructed view into the reaction chamber through the center of the cone, thus enabling more reliable detection of color changes.

[0059] The object underlying the invention is also achieved by a heating device for heating a reaction vessel for isothermal heating for LAMP analysis to detect the presence of DNA and / or RNA in a sample, in particular a reaction vessel according to the present invention, comprising at least two reaction chambers, a receptacle for the reaction vessel which can be inserted into the receptacle in an insertion direction, so that the reaction vessel can be fixed in a defined position, wherein an electrical heating element is formed in the area of ​​the reaction chambers of the reaction vessel, which is suitable and intended for heating to and maintaining a predeterminable temperature by establishing an electrical connection with a voltage supply, wherein a control device is formed by which the heating process can be started automatically when the reaction vessel is inserted into the receptacle and can be terminated after a predeterminable time has elapsed.A control device is preferably provided for controlling and, if necessary, regulating the heating element, which is suitable and intended for heating the heating element to the predefinable temperature and for maintaining it at the predefinable temperature for a predefinable period of time.

[0060] Preferably, a temperature sensor is incorporated whose signal can be used to control the heating and isothermal heating process to a predefinable temperature. Alternatively, the electrical heating element is made of a PTC material whose final temperature corresponds to the temperature required for the LAMP analysis.

[0061] In the case of the design of a temperature sensor, for example a thermistor whose electrical resistance changes depending on the temperature of the thermistor, different target temperatures can be specified, so that the heating device can be used preferably for different LAMP reactions for the detection of different DNA and / or RNA. 40099 / 4361 / PTWO 7 November 2025

[0062] 11

[0063] The temperature sensor is preferably made of a material with a positive temperature coefficient (PTC) and is in particular installed in direct thermal contact with the heating element.

[0064] The use of a PTC material, i.e., a material with a positive temperature coefficient (PTC), as an electrical heating element has the advantage that it automatically regulates itself to the final temperature during operation. This eliminates the need for complex control systems and simplifies the design of the corresponding control device.

[0065] Preferably, a contact switch is designed that starts the heating process upon contact with the reaction vessel. This simplifies the operation of the heating device, as the user only needs to slide the reaction vessel into the holder until it reaches a predetermined temperature. This is preferably achieved by a stop in the holder; in particular, it is preferred that the contact switch rises in a ramp-like manner in the insertion direction, thus simultaneously providing a perceptible stop for the reaction vessel, fixing it relative to the holder, and thereby defining the position of the reaction vessel and, in particular, the reaction chambers of the reaction vessel relative to the heating element.

[0066] Preferably, an optical status indicator is provided to visually display the status of the heating process. Preferably, the optical status indicator is designed as an LED (light-emitting diode) whose color is adjustable, so that the status of the heating process can be indicated by the color of the light emitted by the LED. For example, the LED illuminates in a first color when the heating device heats up to the preset temperature, in a second color when the temperature is maintained at the preset temperature, and in a third color when the heating process is complete and the analysis result can be read.

[0067] Furthermore, the underlying problem is solved by a system for performing a LAMP analysis to detect the presence of DNA and / or RNA in a sample, comprising a sample preparation device according to the present invention and a reaction vessel according to the present invention. Preferably, the system further comprises a heating device according to the present invention.

[0068] The sample preparation device described here works advantageously in conjunction with the reaction vessel described here. The reaction vessel, in turn, is preferably designed to correspond to the heating device. The above for the individual elements 40099 / 4361 / PTWO 7 November 2025

[0069] 12

[0070] The details and advantages disclosed above (sample preparation device, reaction vessel, and heating device) can be transferred and applied to the system according to the present invention. The synergies resulting from the interaction of the elements have already been discussed above.

[0071] Preferably, the access port of the reaction vessel has a centering aid which, in conjunction with a positioning collar of the sample preparation device, allows a hollow cannula of the sample preparation device, connected to the eluate reservoir, to be positioned relative to the first membrane of the access port. This enables optimal positioning of the hollow cannula relative to the self-sealing first membrane of the access port. This ensures that the eluate is forced exclusively into the fluid channel of the reaction vessel.

[0072] Preferably, the access port of the reaction vessel has a stop that, when a sample preparation device is attached to the access port, allows the eluate reservoir of the sample preparation device to be elastically and / or deformable, thus building up pressure in the eluate reservoir, particularly when the eluate reservoir is moved against the stop in the direction of actuation. In particular, a movement to advance a hollow cannula of the sample preparation device, which is in fluid communication with the eluate reservoir at one end and whose second end, opposite the first, is guided through the first membrane into the access port, can simultaneously deform the eluate reservoir and thus increase the pressure in the eluate reservoir, forcing the eluate through the hollow cannula into the access port and thus into the fluid channel system of the reaction vessel.

[0073] Furthermore, a method for LAMP analysis to detect the presence of DNA and / or RNA in a sample using a system according to the present invention is proposed, comprising the following steps:

[0074] • Providing the sample;

[0075] • Contacting the sample with the lysis buffer in the transfer vessel of the sample preparation device;

[0076] • Building up pressure by which the formed lysate is conveyed through the purification unit into the eluate reservoir;

[0077] • Transfer of the eluate under pressure from the sample preparation device through the access port into the reaction vessel, so that the eluate enters the reaction chambers; 40099 / 4361 / PTWO 7 November 2025

[0078] 13

[0079] • Heating the reaction chambers to a temperature adapted for the LAMP reaction of the DNA or RNA to be tested; and

[0080] • Maintaining the reaction chambers at the adjusted temperature for the duration required for the LAMP reaction to proceed.

[0081] After holding the sample at the adapted temperature, LAMP amplification of DNA and / or RNA present in the sample occurs. The presence of DNA can preferably be detected optically, for example by a color change. Preferably, the heating and holding of the reaction chambers at the adapted temperature takes place in a heating device according to the present invention.

[0082] In the present invention, the term "one," "a," or "an" does not refer to a specific number of the components thereby designated, but explicitly includes the possibility that the claimed subject matter may comprise one or more additional components besides the single component. Thus, the term is also to be understood as "one or more." If, on the other hand, the number of specified components is meant to be one, this is indicated by "exactly one."

[0083] The embodiments described above can be combined by a person skilled in the art in any way possible within the framework of the teaching of the invention, even deviating from the references to the claims.

[0084] Brief description of the drawings

[0085] They show schematically:

[0086] Fig. 1: an example of a reaction vessel in top view;

[0087] Fig. 2: the example of the reaction vessel in a partially exploded cross-sectional drawing;

[0088] Fig 3: an example of a sample preparation device in section;

[0089] Fig. 4: a detail of the example of the sample preparation device in a first position;

[0090] Fig. 5: Detail of the sample preparation device example in a second position;

[0091] Fig. 6: a cross-section along the line labelled Vl-Vl in Fig. 5;

[0092] Fig. 7: an example of a heating device in top view;

[0093] Fig. 8: an example of a heating device in cross-section;

[0094] Fig. 9: An example of a system for analyzing samples in a perspective view; 40099 / 4361 ZPTWO 7 November 2025

[0095] 14

[0096] Fig. 10: schematically depicts the process of an analysis method according to the present invention.

[0097] Detailed description of the drawings

[0098] The embodiments shown here are merely examples of the present invention and should therefore not be interpreted as limiting. Alternative embodiments considered by a person skilled in the art are likewise covered by the scope of protection of the present invention.

[0099] Fig. 1 shows a top view of an example of a reaction vessel 100, and Fig. 2 shows a cross-sectional view of an example of a reaction vessel 100. Unless explicitly stated otherwise, both figures are described together below. The reaction vessel 100 comprises a vessel body 102, which can be manufactured, for example, by injection molding from a plastic material. The vessel body 102 is plate-shaped.

[0100] The reaction vessel 100 has an access port 104 through which a sample fluid can be introduced into a fluid channel system 108 via a self-sealing first membrane 106, for example, made of a silicone material, in particular a poly(organo)siloxane material. For this purpose, the first membrane 106 is pierced with a needle or hollow cannula (not shown) and a fluid to be analyzed is forced into the fluid channel system 8. A centering aid 107, designed as a circumferential collar, assists in positioning a correspondingly designed sample preparation device (not shown) relative to the access port 104. Furthermore, the access port 104 has a stop 105 against which parts of a sample preparation device (not shown) can abut. Preferably, the stop 105 is designed as a lip in the first membrane 106.By pressing an eluate reservoir in the sample preparation device against the stop 105, it can be deformed, thereby increasing the pressure in the eluate reservoir, which can then be used to convey an eluate present in the eluate reservoir.

[0101] The fluid channel system 108 comprises a main channel 110 that connects the access port 104 to several secondary channels 112. Each secondary channel 112 terminates in a reaction chamber 114. Each reaction chamber 114 contains a reaction solution 116 in a lyophilized (freeze-dried) form. Several reaction chambers 114 may contain the same reaction solution 116; however, it is preferred that each reaction chamber 114 contains a different reaction solution 116 in order to be able to test for different DNA / RNA and, if necessary, to carry out control reactions. 40099 / 4361 / PTWO 7 November 2025

[0102] 15. The reactions taking place in reaction chambers 114 with the reaction solution 116 are described below.

[0103] When the fluid channel system 108 is filled through the access port 104, the fluid to be analyzed is forced under pressure into the fluid channel system 108, displacing the air previously present in the fluid channel system 108. The air exits the fluid channel system 108 via a second membrane 118 (see Fig. 2). The second membrane 118 is designed to allow air to pass through, but not the fluid present in the reaction chamber 114. Preferably, the second membrane 118 is designed as a hydrophobic, narrow-pored membrane (pore size, for example, 0.2 pm [micrometers]), in particular made of a synthetic material, especially in the form of a film. For clarity, the second membrane 118 is shown above the reaction vessel 100 in Fig. 2, but it is bonded to the upper surface 120 of the vessel, in particular by gluing and / or welding, so that it seals off the reaction chambers 114 for the fluid to be analyzed.The second membrane 118 is preferably white and opaque to ensure a standardized background and good color contrast when optically evaluating the reaction result of the samples.

[0104] Upon entering a reaction chamber 114, the lyophilized reaction solution 116 present in the reaction chamber 114 is dissolved. The reaction solutions 116 are loaded with pathogen-specific primers to enable the amplification of specific DNA and / or RNA and thus the detection of various pathogens in the fluid to be analyzed.

[0105] The reaction chambers 114 taper conically towards the top 120 of the reaction vessel 100. This reduces the volume of the reaction chambers 114 and thus the sample volume to be analyzed. Preferably, the reaction chambers 114 have a volume of approximately 20 to 50 pl [microliters].

[0106] On one of the undersides 122 opposite the top 120, the reaction vessel 100 is closed by a base 124 made of a transparent material. The base 124 has viewing cones 126 whose position and dimensions correspond to the reaction chambers 114 and which preferably project partially into the reaction chambers 114. The viewing cones 126 facilitate the visual perception of the test result in the reaction chamber 114, which is described in more detail below, since any air bubbles that may be present, which can also arise from the heating that aids the reactions and the ongoing chemical reaction, are contained at the edge of the 40099 / 4361 / PTWO 7 November 2025

[0107] 16

[0108] The cone collects the air bubbles so that the reading can be taken through the cone base 128 of each viewing cone 126 without interference from the air bubbles. The base 124 is bonded to the base body 102.

[0109] The reaction vessel 100 and the base body 102 are plate-shaped. This means, in particular, that the top surface 120 and the bottom surface 122 each have an area that is larger than the area of ​​the edges 125 connecting the top surface 120 and the bottom surface 122, in particular larger than the sum of the areas of the edges 125, and in particular at least by a factor of five larger than the sum of the areas of the edges 125.

[0110] Fig. 3 schematically shows a cross-sectional view of a sample preparation device 200. This device comprises a sample receiving chamber 202 into which a sample, for example, taken via a cotton ball or a spatula and which may include saliva, blood, stool, urine, tissue, and / or aqueous fluids containing RNA or DNA, is wiped. For this purpose, the sample receiving chamber 202 is detachable from the rest of the sample preparation device 200 and has a lysis buffer that breaks down the components of the sample after it is added to the sample receiving chamber 202. The sample receiving chamber 202 can be connected to the transfer vessel 206 via bayonet fittings (not shown), which, once the connection is established, prevent unintentional disconnection.

[0111] After appropriate lysis in the sample receiving chamber 202, the resulting lysate is transferred to a transfer vessel 206. For this purpose, pressure is generated by connecting the sample receiving chamber 202 to the transfer vessel 206, forcing the lysate through a filtration column 208 filled with packing material (not shown) and a filter 210. The filter 210 serves both to mechanically support the packing material of the filtration column 208 and to provide additional filtration of the lysate. After passing through the filter 210, the resulting eluate is stored in the eluate reservoir 212. The transport of the sample from the transfer vessel 206 through the filtration column 208 and the filter 210 occurs exclusively via the connection between the sample receiving chamber 202 and the transfer vessel 206.

[0112] Instead of transferring the lysate from the sample receiving chamber 202, it is also preferably possible to transfer a corresponding lysate from another source into the transfer vessel 206 and then, if necessary, to design the upper part of the sample preparation device 200 accordingly without a lysis buffer. 40099 / 4361 ZPTWO 7 November 2025

[0113] 17

[0114] The transfer vessel 206, together with a purification unit 214 consisting of a filtration column 208 and a filter 210, a first cylinder 218, and a second cylinder 220, forms a piston 216. After being unlocked, for example by rotating the piston 216 relative to a housing 201 of the sample preparation device 200 (described in more detail below with reference to Figures 4 to 6), the piston 216 is displaceable against the housing 201, which serves as a guide for the piston 216. The first cylinder 218 and the second cylinder 220 can also be formed as a single piece or as a single cylinder.

[0115] As the piston 216 moves relative to the housing 201, at least part of the second cylinder 220 is pushed outwards over the eluate reservoir 212 (see Fig. 4, which shows the piston 216 before movement in the direction of movement 217, and Fig. 5, which shows the piston 216 upon reaching a first position). The eluate reservoir 212 has an opening at its top (not shown) through which pressure generated when the second cylinder 220 is moved relative to the eluate reservoir 212 is released via a vent opening (not shown) in the second cylinder 220. The eluate reservoir 212 preferably also has further openings, perforations, or pores, particularly on its circumference, to allow venting through the vent opening (not shown).

[0116] When the second cylinder 220 reaches a first position (see Fig. 5), it completely covers the outer circumference of the eluate reservoir 212. As shown in Fig. 5, venting of the eluate reservoir 212 is no longer possible, as the reservoir is hermetically sealed around its circumference by a seal 217. At this point, the entire sample has been forced through the purification unit 214 and is now present as eluate in the eluate reservoir 212.

[0117] If the piston 216 is moved further in the actuation direction 217, the eluate reservoir 212 is now moved together with the piston 216 in the actuation direction 217. The eluate reservoir 212 is carried along by the piston 216 from this position. If the eluate reservoir 212 is in contact with the bottom, for example against a suitably designed access port 104, the eluate reservoir 212 is deformed, so that an overpressure now builds up in the eluate reservoir 212, which can be used to pump the eluate.

[0118] Figures 4 and 5 will now be described. These figures will be described together unless explicit reference is made to one of them. 40099 / 4361 / PTWO 7 November 2025

[0119] 18

[0120] Figures 4 and 5 show a detail of the sample preparation device 200 in two different positions. Visible are the second cylinder 220, the eluate reservoir 212 with hollow cannula 224, and a release unit 232, the function of which is described in more detail below, particularly with reference to Figure 6. The eluate reservoir 212 is open at the top and sealed circumferentially by a seal 213, for example, an O-ring. The second cylinder 220 has a vent opening 230, which is formed as a corresponding recess on the inside of the hollow second cylinder 220.

[0121] In the first position (Fig. 4) of piston 217, prior to movement of piston 216 in the actuation direction 217, piston 216 can be unlocked by a rotational movement 244, shown particularly below with reference to Fig. 6. Only after unlocking can piston 216 be pushed over the eluate reservoir 212 with the second cylinder 220. The eluate reservoir 212 has an opening 234 at the top through which the eluate enters the reservoir 212. In the first position, the space above the eluate reservoir 212 can be vented via the vent opening 230, so that when cylinder 218 is moved to the second position (Fig. 5), a defined pressure build-up occurs, and pressure can be released via the vent opening 230 until then. In the second position, the second cylinder 220 strikes the eluate reservoir 212. If the second cylinder 220 is moved further in the actuation direction 217, it takes the eluate reservoir 212 with it from the second position (Fig. 5).The position of the O-ring 213 is chosen such that, upon reaching the second position, there is already an increased pressure in the eluate reservoir 212, which can be used to transport the eluate from the eluate reservoir 212.

[0122] With reference in particular to Fig. 6, the release unit 232 will now be described. This unit comprises a running body 236, which is guided in a running groove 238 that connects a locking recess 240 and a release recess 242. By means of a spring-loaded detent device (not shown in detail), the running body 236 can be detented in the locking recess 240, so that after detent locking, the piston 216 and thus the second cylinder 220 is moved in the actuation direction 217. The release unit 232 and / or the housing 201 is designed such that movement of the piston 216 relative to the housing 201 is permitted.

[0123] By a rotational movement 244 of the cylinder 218 relative to the piston 216, as also indicated in Figures 4 and 5, the running body 236 can be moved between release recess 242 and locking recess 240 and then secured in the locking recess 240. The detent movement 246 of the detent areas 248 in the radial direction is also shown in Figures 4 to 6. 40099 / 4361 ZPTWO November 7, 2025

[0124] 19

[0125] With further reference to Fig. 3, the transfer of the eluate will now be described. To transfer the eluate into a reaction vessel (not shown here), in particular a reaction vessel 100 as shown in Figures 1 and 2 and described above, the sample preparation device 200 is positioned over the reaction vessel. In the case of the reaction vessel 100 as described above, a positioning collar 222, which forms a lower end of the sample preparation device 200 below the eluate reservoir 212, is positioned in the centering aid 107 of the access port 104 of the reaction vessel 100.

[0126] The sample preparation device 200 has a hollow cannula 224 which, when the piston 216 and the cylinder 218 are positioned in the second position described above, is pushed downwards against the action of a compression spring 226 by pressing on the piston 216 of the sample preparation device 200 in an actuation direction 217 until the hollow cannula 224 protrudes above a lower edge 228 of the positioning collar 222 and thereby pierces the self-sealing first membrane 106 of the access port 104 of the reaction vessel 100. The hollow cannula 224 is positioned precisely above the center of the self-sealing first membrane 106 by the interaction of the positioning collar 222 and the centering aid 107. The pressure on piston 216 causes piston 216 and thus the second cylinder 220 to be moved further downwards, whereby the eluate reservoir 212 is pressed against the stop 105 of the access port 104 and is thereby deformed.The resulting increased pressure in the eluate reservoir 212 transports the eluate under pressure through the hollow cannula 224 and the access port 104 into the fluid channel system 108 of the reaction vessel 100 (see Figs. 1 and 2). There it is distributed to the reaction chambers 114 as described above.

[0127] After the pressure on the piston 216 is released, the compression spring 226 causes a return movement, which moves the piston 216, the second cylinder 220, and the eluate reservoir 212 back into the second position (Fig. 5). As a result, the hollow cannula 224 no longer protrudes beyond the lower edge 228 of the positioning collar 222; the tip of the hollow cannula 224 lies within the positioning collar 222. This prevents injury to a user of the sample preparation device 200.

[0128] The purification unit 214, formed by filtration column 208 and filter 210, is adapted in its physical and chemical configuration to the type of analysis to be performed. This also applies to the corresponding reaction solutions 116 in the reaction chambers 114. Thus, appropriate chemical reactions and physical filtration processes take place in filtration column 208, which ensure that the eluate meets the requirements of 40099 / 4361 ZPTWO, November 7, 2025.

[0129] 20

[0130] Contains reactants required for the analytical reaction in reaction chamber 114, while other components of the sample and / or the lysis are filtered out.

[0131] Fig. 7 shows a heating device 300 for heating a reaction vessel, in particular a reaction vessel 100 as shown in Figs. 1 and 2 and described above, in a top view, while Fig. 8 shows the heating device 300 in section. Figs. 3 and 7 are subsequently described together unless explicit reference is made to one of the figures.

[0132] The heating device 300 comprises a housing 302, which has a receptacle 304 for a reaction vessel, in particular a reaction vessel 100 according to the present invention and as described above. The receptacle 304 corresponds in shape and dimensions to the reaction vessel 100 to be heated. When the reaction vessel 100 is inserted in the insertion direction 305 (see Fig. 7), a contact switch 306 is actuated, which initiates a heating process.

[0133] For this purpose, a heating element 308, over which the reaction vessel 100 can be positioned, particularly in the area of ​​its reaction chambers 114, is electrically connected to a power supply 310 (see Fig. 8), in particular a battery, an accumulator, or a power supply unit connectable to a mains power supply. The heating element 308 is controlled by a control unit 312, which monitors and regulates the heating process. The heating element 308 is preferably a thick-film heating element. A thermocouple (not shown) is located below the heating element 308 as a temperature sensor, for example, made of a material with a positive or negative temperature coefficient (NTC or PTC), which monitors the temperature. The change in the resistance of the temperature sensor due to the changing temperature is monitored and regulated electronically. The logic thresholds can be adjusted, in particular, by hardware programming.

[0134] The heating device 300 also includes an optical status indicator 314, which in this example is designed as an LED. The status indicator 314 displays a first color, for example blue, during heating; a second color, for example red, while maintaining the temperature; and a third color, for example green, after the preset time for the reactions has elapsed. This allows the user of the heating device 300 to monitor the progress of the heating process. 40099 / 4361 ZPTWO November 7, 2025

[0135] 21

[0136] The heating device 300 serves to heat the reaction vessel 100 to a predetermined temperature, preferably in the range of 65 to 85°C [degrees Celsius]. The temperature is then maintained for a predetermined time. During this time, reactions take place in the reaction chambers 114 of the reaction vessel 100 as described below.

[0137] Figure 9 shows a perspective view of System 400 for sample analysis. System 400 comprises a reaction vessel 100 as described and shown in this document, a sample preparation device 200 as described and shown in this document, and a heating device 300 as described and shown in this document. Through the interaction of these elements, samples, such as saliva, blood, stool, urine, tissue, and aqueous fluids containing RNA or DNA, can be examined for the presence of various pathogens, particularly viruses or bacteria.

[0138] Figure 10 illustrates the process of analyzing a sample using the system 400. In a first step 500, a sample 502 is taken from a subject. The sample 502 is taken from the subject's nose using a sampler 504. In a second step 506, the cover 204 is removed from the sample receiving chamber 202 of the sample preparation device 200. In a third step 508, the sample 502 is placed into the sample receiving chamber 202 using the sampler 504 and brought into contact with the lysate.

[0139] In a fourth step 510, after a predetermined time has elapsed for the lysis to take place, the piston 216 is unlocked by a rotation against the housing 201, moved to the second position, and thereby, as described above, the lysate is forced through the purification unit 214 (not shown here), purified there, and then forced as eluate into the eluate reservoir 212 (not shown here).

[0140] In the fifth step 512, the eluate is forced, as described above, by pressing the piston 216 of the sample preparation device 200 in the actuation direction 217 through the access port 104 into the fluid channel system of the reaction vessel 100 (not shown). In a sixth step 514, the appropriately filled reaction vessel 100 is inserted into the receptacle 304 of the heating device 300, and the reaction vessel 100 is heated. In a seventh step 516, the reaction vessel 100 is held at a predetermined temperature for a predetermined time, during which LAMP amplification takes place in the reaction chambers (not shown). In 40099 / 4361 / PTWO 7 November 2025

[0141] 22 In an eighth step 518 after the specified time has elapsed, the LAMP amplification is completed and the reaction vessel 100 can be removed from the heating device 300 for optical verification of the reaction result.

[0142] The system 400 presented here for performing LAMP amplification and detecting the presence of DNA and / or RNA in a sample consists of the elements of the sample preparation device 200, a reaction vessel 100, and optionally a heating device 300. The system 400 allows for the simple and rapid detection of the presence of DNA and / or RNA in a sample, for example, to detect the presence of a virus in the sample.

[0143] 40099 / 4361 / PTWO November 7, 2025

[0144] 23

[0145] List of reference symbols

[0146] 100 reaction vessels

[0147] 102 vessel bodies

[0148] 104 access ports

[0149] 105 stop

[0150] 106 self-sealing first membrane

[0151] 107 Centering aid

[0152] 108 Fluid channel system

[0153] 110 Main Channel

[0154] 112 Secondary channel

[0155] 114 reaction chamber

[0156] 116 reaction solution

[0157] 118 second membrane

[0158] 120 Top

[0159] 122 Underside

[0160] 124 Floor

[0161] 125 edge

[0162] 126 Visible cone

[0163] 128 Conical base

[0164] 200 Sample preparation device

[0165] 201 cases

[0166] 202 Sample recording room

[0167] 204 lids

[0168] 206 Transfer vessel

[0169] 208 Filtration column

[0170] 210 filters

[0171] 212 Ela reservoir

[0172] 213 Seal

[0173] 214 Purification unit

[0174] 216 pistons

[0175] 217 Direction of action

[0176] 218 first cylinder

[0177] 220 second cylinder

[0178] 222 Positioning collar

[0179] 224 Hollow cannula 40099 / 4361 ZPTWO November 7, 2025

[0180] 226 Compression spring

[0181] 228 lower edge of the positioning collar

[0182] 230 Venting of the eluate reservoir

[0183] 232 Release unit

[0184] 234 Opening

[0185] 236 barrels

[0186] 238 Running channel

[0187] 240 locking recess

[0188] 242 Release in-depth

[0189] 244 Rotational movement

[0190] 300 heating device

[0191] 302 Cases

[0192] 304 recording

[0193] 305 Insertion direction

[0194] 306 contact switches

[0195] 308 Heating element

[0196] 310 Power supply

[0197] 312 Control unit

[0198] 314 optical status indicators

[0199] 400 System for the analysis of samples

[0200] 500 first step

[0201] 502 Sample

[0202] 504 samplers

[0203] 506 second step

[0204] 508 third step

[0205] 510 fourth step

[0206] 512 fifth step

[0207] 514 sixth step

[0208] 516 seventh step

[0209] 518 eighth step

Claims

40099 / 4361 / PTWO November 7, 2025 25 Patent claims 1. Sample preparation device (200) for preparing a sample for carrying out a LAMP analysis to detect the presence of DNA and / or RNA in the sample, comprising • a sample receiving chamber (202) which preferably includes a lysis buffer; • a purification unit (214); and • an eluate reservoir (212) wherein the purification unit (214) is part of a flask (216), wherein the sample receiving chamber (202) is detachably connectable to the flask (216), so that when the sample receiving chamber (202) is connected to the flask (216) a pressure can be built up in the flask (216), by which a lysate, preferably formed in the sample receiving chamber (202) by reaction of the sample with the lysis buffer, can be conveyed through the purification unit (214) into the eluate reservoir (212) by forming an eluate.

2. Sample preparation device (200) according to claim 1, characterized in that the piston (216) is displaceable relative to the eluate reservoir (212) in an actuation direction (217).

3. Sample preparation device (200) according to claim 2, characterized in that a release unit (232) is provided which then enables a displacement of the piston (216) in the actuation direction (217) towards the eluate reservoir (212) when the release unit (232) has been moved into an unlocked position.

4. Sample preparation device (200) according to claim 3, characterized in that the release unit (232) can be moved into the unlocking position by a rotational movement (244) of the piston (216).

5. Sample preparation device (200) according to one of claims 2 to 4, characterized in that, during the movement of the piston (216) against the eluate reservoir (212), a cylinder (220) can be slid externally over the eluate reservoir (212). 40099 / 4361 / PTWO November 7, 2025 26 6. Sample preparation device (200) according to one of the preceding claims, characterized in that the purification unit (214) comprises a filtration column (208) and a filter which are adaptable to the sample and the DNA and / or RNA to be detected.

7. Reaction vessel (100) for providing a sample for LAMP analysis to detect the presence of DNA and / or RNA in the sample, comprising an access port (104) for adding the sample, which is present as an eluate, into a fluid channel system (108) connected to the access port (104), which connects the access port (104) to at least two reaction chambers (114), wherein each reaction chamber (114) contains a reaction solution (116) in lyophilized form specific for the DNA and / or RNA to be detected, wherein at least the reaction chambers (114) are closed at a top (120) of the reaction vessel (100) by a second membrane (118) which is permeable to air but not to the eluate.

8. Reaction vessel (100) according to claim 7, characterized in that the access port (104) is closed by a first membrane (106) which is self-closing.

9. Reaction vessel (100) according to claim 7 or 8, characterized in that the reaction chambers (114) are conically shaped.

10. Reaction vessel (100) according to one of claims 7 to 9, characterized in that the reaction vessel (100) has a transparent bottom (124) on a bottom (122) opposite the top (120).

11. Reaction vessel (100) according to claim 10, characterized in that the bottom (124) in the area of ​​the reaction chambers (114) has a sight cone (126) in each reaction chamber (114) which tapers conically towards the top (120).

12. Heating device (300) for heating a reaction vessel for isothermal heating for LAMP analysis to detect the presence of DNA and / or RNA in the sample, in particular a reaction vessel (100) according to one of claims 7 to 11, with at least two reaction chambers (114), comprising a receptacle (304) for the reaction vessel (100), which is in a 40099 / 4361 / PTWO November 7, 2025 27 The insertion direction (305) into the receptacle (304) is slidable so that the reaction vessel (100) can be fixed in a defined position, wherein an electrical heating element (308) is formed in the area of ​​the reaction chambers (114) of the reaction vessel (100), which is suitable and intended for heating to and maintaining a predeterminable temperature by establishing an electrical connection with a voltage supply (310), wherein a control device (312) is formed by which the heating process can be started automatically when the reaction vessel (100) is inserted into the receptacle (304) and can be terminated after a predeterminable time.

13. Heating device (300) according to claim 12, characterized in that the electrical heating element (308) is made of a PTC material whose final temperature corresponds to the temperature required for the LAMP analysis.

14. Heating device (300) according to one of claims 12 to 13, characterized in that a contact switch (306) is provided which starts the heating process upon contact with the reaction vessel (100).

15. Heating device (300) according to one of claims 12 to 14, characterized in that an optical status indicator (314) is provided by which the status of the heating process can be visually indicated.

16. System (400) for performing a LAMP analysis to detect the presence of DNA and / or RNA in the sample, comprising a sample preparation device (200) according to any one of claims 1 to 6 and a reaction vessel (100) according to any one of claims 7 to 11.

17. System (400) according to claim 16, further comprising a heating device (300) according to any one of claims 12 to 15.

18. System (400) according to claim 16 or 17, characterized in that the access port (104) of the reaction vessel (100) has a centering aid (107) by which, in conjunction with a positioning collar (222) of the sample preparation device (200), a system connected to the eluate reservoir (212) 40099 / 4361 / PTWO November 7, 2025 28 The hollow cannula (224) of the sample preparation device (200) can be positioned relative to the first membrane (106) of the access port (104).

19. System (400) according to one of claims 16 to 18, characterized in that the access port (104) of the reaction vessel (100) has a stop (105) by which, when the sample preparation device (200) is placed on the access port (104), the eluate reservoir (212) of the sample preparation device (200) can be deformed to build up pressure in the eluate reservoir (212).

20. Method for LAMP analysis for the detection of the presence of DNA and / or RNA in a sample using a system (400) according to any one of claims 16 to 19, comprising the following steps: • Providing the sample; • Contacting the sample with the lysis buffer in the transfer vessel (202) of the sample preparation device (200); • Building up pressure by which the formed lysate is conveyed through the purification unit (214) into the eluate reservoir (212); • Transfer of the eluate under pressure from the sample preparation device (200) through the access port (104) into the reaction vessel (100), so that the eluate enters the reaction chambers (114); • Heating the reaction chambers (114) to a temperature adapted for the LAMP reaction of the DNA or RNA to be tested; and • Maintaining the reaction chambers (114) at the adjusted temperature for a duration required for the LAMP reaction to proceed.

21. Method according to claim 20, characterized in that the heating and holding of the reaction chambers at the adapted temperature is carried out in a heating device (300) according to one of claims 12 to 15.

Images