In-vitro diagnostic test systems, IVD test devices and methods for multiplexed diagnostic assays with improved efficiency and eco-friendliness

Abstract

This invention enables increased throughput of diagnostic tests, doubling, tripling, or even further the number of tests per cartridge, and is therefore considered highly environmentally and ecologically friendly. Simultaneously, it provides multiple potentially life-saving test results for one or more patients. The present invention relates to an in vitro diagnostic (IVD) testing system (1a, 1b) for performing multiple diagnostic assays, wherein the IVD testing system (1a, 1b) comprises: a test carrier (2) including a sample application port (3) configured to receive a sample fluid (30); a test region (4) including a common recessed assay membrane region (4a) and a sample release port (4b) for releasing at least one portion (30, 30a) of the sample fluid (30) into the common recessed assay membrane region (4a); and a microfluidic sample channel system (5) configured to guide the at least one portion (30, 30a) of the sample fluid (30) from the sample application port (3) to the sample release port (4b); the IVD testing system (1a, 1b) further comprises: a first assay membrane (6) positioned in the common recessed assay membrane region (4a) and configured to release the sample fluid (30) from the sample application port (3) into the sample release port (4b). The sample release port (4b) receives a first portion of the at least one (30, 30a) of the sample fluid (30) and indicates at least one first analyte (31a) in the first portion of the at least one (30, 30a) of the sample fluid (30); and a second assay membrane (7) is positioned adjacent to the first assay membrane (6) in the common recessed assay membrane region (4a) and is configured to receive a second portion of the at least one (30, 30a) of the sample fluid (30) from the sample release port (4b) and indicate at least one second analyte (31b) in the second portion of the at least one (30, 30a) of the sample fluid (30) and / or indicate at least one first analyte (31a) in the second portion of the at least one (30, 30a) of the sample fluid (30) with a sensitivity range different from that of the first assay membrane (6).

Description

Technical Field

[0001] This invention relates to an in vitro diagnostic (IVD) testing system, an IVD testing device, and a method for performing multiple diagnostic assays to detect several parameters or at least one parameter (specifically including but not limited to cardiac parameters) within different measurement ranges (i.e., sensitivity ranges). This invention allows for increased throughput of diagnostic assays and can double, triple, or even more the number of assays per test cartridge, and is considered very environmentally friendly due to its significant reduction in waste. Simultaneously, it can provide multiple potentially life-saving test results for one or more patients in a short period of time. Background Technology

[0002] In medical diagnostics, providing test results immediately or within a very short time is often crucial, especially when patients are in life-threatening health conditions. To identify the severity of acute health conditions and / or circumstances, it may often be necessary to simultaneously and reliably determine several parameters and / or properties of a patient's sample, enabling decisions to be made regarding the application of appropriate and effective treatment. Furthermore, it may be necessary to provide rapid test results for several patients in urgent need of medical assistance simultaneously. Therefore, point-of-care (POC) medical testing used in emergency rooms, medical practices, and other settings requires the efficient delivery of reliable test results for multiple parameters, such as the qualitative or quantitative determination of analytes in fluids / liquids from one or more patients' samples.

[0003] In the field of in vitro diagnostics, medical point-of-care (POC) tests for the qualitative and / or quantitative determination of analytes in liquid samples are typically based on immunoassays that can utilize lateral flow assay strips. In most cases, lateral flow assay (LFA) strips, including porous immunoassay membranes (e.g., nitrocellulose membranes) or paper (e.g., cellulose), are used for such applications, such as as assay membranes and / or functional pads, which have the ability to passively deliver fluid via capillary forces. Typically, such LFA strips are coated with a capture reagent for capturing and detecting the target analyte. This capture and detection zone is also referred to as a test line or test zone.

[0004] To provide test results for multiple parameters simultaneously, multiple point-of-care (POC) tests can be used. These multiple POC tests allow the simultaneous identification of more than one analyte from the same sample in a single measurement run using a single test vehicle. This type of test is often referred to as a "kit test." The POC platform should offer the possibility of performing both single and multiple tests on the same system, enabling healthcare professionals to perform measurements as conveniently and quickly as possible. From a manufacturer's perspective, it is ideal to achieve this platform's flexibility in both single and kit tests without requiring significant changes to the design of the test vehicle and instrument, or the use of the same sample volume.

[0005] For example, Figures 1a and 1b illustrate the concept of a known point-of-care (POC) test using an immunoassay membrane 6 (also referred to as the “assay membrane”). A sample fluid 30, such as a whole blood sample in this example, is received from the patient. As shown in Figure 1a, the sample fluid 30 is indicated to contain a target analyte 31, against which the sample fluid 30 should be tested. Plasma 30a is extracted from the whole blood sample 30, for example, by centrifugation. Plasma 30a can be considered a component of the whole blood sample 30 and, after separation (i.e., extraction from the whole blood sample 30), contains analyte 31. The plasma 30a is then mixed with drying reagents 32 and 33, which are designed to specifically conjugate to the binding site of analyte 31 and thus label analyte 30a. Drying reagents 32 and 33 include a detection reagent 32 and a capture reagent 33. The detection reagent 32 includes a detection antibody for conjugation to the binding site of analyte 31. The detection reagent 32 includes and / or is coupled to a detection element for providing a detection signal, which may, for example, correspond to a fluorescent label and / or JG9-latex beads. The capture reagent 33 includes a capture antibody 33 for coupling to another binding site of the analyte 31. The capture reagent 33 includes and / or is coupled to a capture element for realizing / performing a capture process on the assay membrane 6, which may, for example, be biotinylated.

[0006] Figure 1b illustrates the capture process of the labeled / labeled sandwich analyte 31c and the detection principle of the LFA strip 6, which corresponds to and / or includes the assay membrane 6 indicated in Figure 1b. The test lines 37 of the assay membrane 6 are coated with analyte-specific antibodies for the specific capture of the target analyte 31. The assay membrane 6 has an overall flow axis 6a, a flow initiation side 6b, and a flow termination side 6c. The flow initiation side 6b and the flow termination side 6c may be defined by the function of the elements on the assay membrane 6. Proper function of the assay membrane 6 is ensured by allowing fluid to enter the assay membrane 6 from the flow initiation side 6b. In other words, the (sample) liquid should be supplied to the assay membrane 6 from the flow initiation side 6b so that the assay can be used correctly. In some cases, the assay membrane 6 may come into contact with a waste pad (which may be waste wool) at the flow termination side 6c (not shown in this figure). The topmost schematic diagram of Figure 1b shows the assay membrane 6 before the application of serum 30a, and... Figure 1bThe middle schematic shows the situation after serum 30a is applied to the assay membrane 6 and the labeled analyte 31c binds to the assay membrane 6. The bottom schematic shows a CCD image of the actual assay membrane 6 as indicated by the middle schematic.

[0007] Plasma 30a containing labeled analyte 31 is applied to the assay membrane 6 at the flow initiation side 6b. At least a portion, particularly a portion and / or component of the plasma 30a, is passively transported substantially along the overall flow axis 6a from the flow initiation side 6b to the flow termination side 6c by capillary force. The assay membrane 6 includes three lines 35, 36, and 37: a control line 35, a calibration line 36, and a capture and test result line 37 (previously referred to as the “test line”). The control line 35 (assay control) is pre-coated with an analyte-specific antibody against the target analyte 31. The calibration line 36 (instrument calibration) is pre-coated with a fluorescent label 32', which is included in a test reagent containing the detection antibody 32. The capture and test result line 37 is coated with a reagent / capture mechanism 34 that captures / binds the capture reagent 32; therefore, this line is the line for performing the capture process. The capture mechanism 34 may, for example, correspond to and / or contain streptavidin.

[0008] Serum 30a is applied to the assay membrane 6 on the flow initiation side 6b and thus first passes through the capture and test result line 37, where, if labeled analyte 31 is present in serum 30a, analyte 31c sandwiched between the two reagents 31, 32 is specifically bound / captured by a capture antibody that binds to the capture mechanism 34 of the capture and test result line 37, i.e., streptavidin in this case. The remainder of serum 30a passes through the calibration line 36, where no further binding occurs, and then through the control line 35, where residual unbound test reagent 32, including the detection antibody, can be at least partially and specifically bound.

[0009] The control line 35 provides a reference for whether serum 31 is mixed with reagents 32 and 33 by capturing unbound test reagent 32. The control line 35 is particularly important if the test result is negative, as in this case, the negative result could be due to the absence of analyte 31, but it could also be due to the absence of test reagent 32 (especially a false negative). The calibration line 36 is used for calibration purposes and provides a pre-coated area with a fluorescent label 32', which is the same fluorescent label 32' used in conjunction with the detection antibody 32 to label analyte 31 for detection.

[0010] Finally, the assay membrane 6 is washed with buffer to remove unbound analytes 31, reagents 32, 33, and / or other molecules, and a CCD image is recorded to read the test results. The bottom schematic of Figure 1b shows this CCD image of the actual assay membrane 6, as indicated by the middle schematic. The three lines 35, 36, and 37 provide visible and / or detectable signals because the bound fluorescent molecules / fluorescent labels 32' are either present on the labeled sandwich analyte 31c on the capture and test result line 37, already present in the pre-coated calibration line 36, or bound to the pre-coated control line 35. Therefore, the bottom schematic of Figure 1b reveals from the visible signal that analyte 31 is present in serum 30a and is clearly well mixed with reagents 32 and 33.

[0011] As shown in Figure 1b, the assay membrane 6 does not provide sufficient space for multiplexing, which would require at least one or even three lines to detect an additional analyte. Furthermore, nonspecific binding events and / or crosstalk between fluorescent labels captured on closely adjacent lines can spoof test results, which is problematic for life-threatening patients who require accurate test results to select appropriate treatment. Therefore, reliable, sensitive, and / or efficient multiplexing is necessary.

[0012] Furthermore, US 2021 / 154662 A1 describes an apparatus for guiding liquid through a porous medium, the apparatus comprising a fluid module rotatable about a center of rotation, and including a fluid chamber and an inflow structure. The porous medium is disposed within the fluid chamber to allow liquid impacting a radially inner portion of the porous medium to flow towards a radially outer portion of the porous medium under centrifugal force. The porous medium is at least partially spaced laterally from the chamber wall of the fluid chamber relative to the flow, such that a fluid connection exists between the radially inner portion of the porous medium and the radially outer portion of the porous medium outside the porous medium. The inflow structure is configured to limit the inflow of liquid into the radially inner portion of the porous medium under centrifugal force to a first flow velocity, wherein the ratio of the first flow velocity to the maximum possible flow velocity through the porous medium is not greater than two.

[0013] US 2011 / 293489 A1 describes systems and methods comprising a microfluidic chip having multiple microfeatures interconnected to provide a constructible fluid delivery system for processing at least one reagent. Intercalations are provided for removably fitting into one or more of the microfeatures of the chip, wherein the intercalations include sites for interacting with the reagent.

[0014] US 2013 / 184188 A1 describes apparatus and methods for performing optical and electrochemical assays, and more specifically, relates to test apparatus having an optically readable microdot array and / or electrochemical detector, and methods for performing microdot array and electrochemical assays using such apparatus. The described apparatus and methods are used for performing point-of-care immunoassays and / or electrochemical assays.

[0015] US 2010 / 120173 A1 describes an immunochromatographic disc assembled with strips, comprising: a base, a cap engaged with the base, and a drain between the strips and the cap disposed on the base, wherein a sampling opening is disposed on the cap directly facing the drain, and the sampling opening is connected to a drain groove formed by a plurality of drain channels disposed on the inner side of the cap; several strip stands are disposed on the base, the position and number of which correspond to the position and number of drain channels disposed on the cap, and the edge of the drain overlaps a sample pad of the strip carried on a stand adjacent to one end of the sampling opening.

[0016] Lutz, Sascha et al., “A fully integrated microfluidic platform for highly sensitive analysis of immunochemical parameters,” ANALYST, September 22, 2017, pp. 4206-4214, XP093171974, UK ISSN: 0003-2654, DOI: 10.1039 / C7AN00547D, describes a microfluidic platform for highly sensitive analysis of immunochemical parameters. Summary of the Invention

[0017] Therefore, it is desirable to provide an IVD testing system, apparatus, and method for reliable, sensitive, and / or efficient multiplex diagnostic assays. Furthermore, it is desirable to provide a platform-based and / or platform-compatible IVD testing system that allows for single-analyte testing and multiplex testing using some, or even almost all, or all of the same components as an IVD testing system. In other words, an IVD testing system is needed that can be used for single-analyte testing and multiplex testing, or that can be easily converted from single-analyte testing to multiplex testing. Further, it is desirable to provide an IVD testing system for multiplex diagnostic assays, manufactured in a manner similar to or identical to that of an IVD testing system for single-analyte testing. Additionally, it is desirable to provide an IVD testing system, apparatus, and method to improve the throughput of diagnostic assays and / or achieve more quantitative diagnostic assays. Further, it is desirable to provide a universal IVD testing system, apparatus, and method for performing multiple diagnostic assays. It is also desirable to provide a compact IVD testing system compatible with rotation-based IVD testing methods.

[0018] At least one of the problems described is addressed by an IVD testing system, an IVD testing apparatus including the IVD testing system, and a method for performing multiple diagnostic assays.

[0019] According to a first aspect of the invention, an IVD testing system for performing multiplex diagnostic assays is provided, wherein the IVD testing system comprises: a test carrier including: a sample application port configured to receive sample fluid; a test region including a shared and / or common recessed assay membrane region (specifically a single shared assay membrane region, and / or which may at least partially correspond to a flat plane that can receive these assay membranes and which may at least partially be limited by recessed walls and recessed relative to an outer surface and / or upper surface and / or a cover plate of the test carrier) and a sample release port (specifically a single sample release port) for releasing at least one portion of the sample fluid into the shared recessed assay membrane region; and a microfluidic sample channel system configured to guide at least one portion of the sample fluid from the sample application port to the sample release port, wherein the IVD The testing system further includes: a first measuring membrane positioned in a shared recessed measuring membrane region and configured to receive at least a first portion of a sample fluid from a sample release port and indicate at least one first analyte in the sample fluid (specifically, in at least a portion of the sample fluid); and a second measuring membrane positioned in the shared recessed measuring membrane region adjacent to the first measuring membrane and configured to receive at least a second portion of a sample fluid from a sample release port and indicate at least one second analyte in the sample fluid (specifically, in the second portion of at least a portion of the sample fluid) and / or indicate at least one first analyte in the sample fluid (specifically, in the second portion of at least a portion of the sample fluid) with a sensitivity range and / or measurement range different from that of the first measuring membrane. In some cases, the sensitivity range may include or be considered as the measurement range.

[0020] This invention allows for increased throughput of diagnostic assays, enabling two, three, or even more tests per cartridge, and is therefore considered highly environmentally friendly (can be understood as a green invention). Simultaneously, it provides multiple potentially life-saving test results for one or more patients. Furthermore, this invention reduces the cost burden on customers running multiple and / or numerous assays in a short period of time.

[0021] A very compact IVD test system is provided for multi-testing, which reduces the number of personalized / customized components in the production of different IVD test systems used for single testing and different multi-testing technologies. This makes the production and use of the IVD test system highly efficient.

[0022] An IVD testing system can be considered a cartridge, such as a centrifugal cartridge that can be used with an IVD testing device (e.g., an IVD testing device including a centrifuge). An IVD testing system allows for reliable, sensitive, and / or efficient multiplex diagnostic assays. An IVD testing system can be platform-based and / or platform-compatible, allowing for single-analyte and multiplex assays using some components of the same parts, specifically using the same test carrier format. Furthermore, the IVD testing system for multiplex assays can be manufactured in a similar or identical manner to the IVD testing system for single-analyte assays. In addition, an IVD testing system allows for increased throughput and / or more quantitative diagnostic assays. An IVD testing system is versatile in performing multiple diagnostic assays.

[0023] Specifically, the IVD testing system allows for multiple diagnostic assays in which at least one analyte can be detected in two different sensitivity ranges and / or measurement ranges to better quantify the analyte in the sample or to determine at least one range. The first and / or second measurement ranges may be, but are not limited to, between about 0.5 mmol / L and 15 mmol / L (specifically, between 2.0 mmol / L and 7.3 mmol / L for potassium, but not limited thereto), and between about 0.05 mg / dL and 50 mg / dL (specifically, between 0.2 mg / dL and 20 mg / dL for creatinine, more specifically between about 0.3 mg / dL and 10 mg / dL). Alternatively or additionally, the IVD testing system allows for multiple diagnostic assays in which at least one first analyte can be reliably detected, and at least one second analyte can be reliably detected, provided the corresponding analyte is present in the original sample fluid. Multiple diagnostic assays (i.e., multiple assays) are performed on two separate assay membranes and are therefore spatially separated from each other.

[0024] The test carrier may consist of one or more parts. If the test carrier includes multiple parts, for example, one part may include a microfluidic sample channel system as a recess, particularly a recessed trace in one part. At least one other part may correspond to a cover plate, specifically a cover plate configured to at least partially close the microfluidic sample channel system. One or more of these parts may include at least a portion of the test area. The test carrier may specifically include a test carrier core sandwiched between at least two cover plates to cover and / or seal the test carrier and / or components contained within the test carrier (such as, for example, drying reagents). One or both of the cover plates may correspond to a membrane and / or sheet. At least one of the membranes and / or sheets may be flexible. At least one of the membranes and / or sheets may provide stability to the test carrier. At least one of the membranes and / or sheets may provide information related to the test type, cartridge type, brand, etc. The test carrier core may be covered only on one side by a cover plate or by several plate-like elements for covering the test carrier core, rather than being sandwiched in the middle. The microfluidic sample channel may correspond to recessed traces in the center and / or core and / or one or more cover plates. Alternatively, the microfluidic sample channel may correspond to a channel, channel system, tunnel, and / or conduit within a block of material parts—in other words, the microfluidic sample channel may be primarily surrounded and / or enclosed by a single element, which may be a one-piece element and in this case, a cover plate may not be required. The test carrier and / or test carrier core may comprise a single piece and / or may be produced by additive manufacturing methods. Alternatively, the test carrier and / or test carrier core may be produced by injection molding. The test carrier and / or test carrier core may comprise two or more parts assembled together, such as two or three or even more plates. For example, a test carrier may comprise three parts assembled together, specifically three plates, with the center and / or core sandwiched between two cover plates. A test carrier may comprise more than three parts assembled together. The test carrier may be at least partially transparent to allow visible light to pass through, and / or for light detectable by a detector, the test carrier may be at least partially transparent. One or all of the components of the test carrier may be at least partially transparent. This allows for the tracking of sample fluid flowing within and / or along the microfluidic sample channel system.

[0025] The test carrier and / or test carrier core and / or any component of the test carrier may be, but is not limited to, at least one of the following materials: polystyrene (PS), polycarbonate (PC), polypropylene (PP), polyethylene (PE), etc. One or more cover plates may be, but is not limited to, at least one of the following materials: polystyrene (PS), polycarbonate (PC), polypropylene (PP), polyethylene (PE), etc. At least one of the described plates may correspond to and / or include foil. Sealing elements may be used to seal the microfluidic sample channel system. Alternatively, sealing elements may not be required to seal the microfluidic sample channel system.

[0026] A sample application port configured to receive sample fluid may correspond to an opening and / or inlet of a microfluidic sample channel system, allowing fluid samples applied to the sample application port to enter the microfluidic sample channel system. For example, the sample application port may correspond to a tapered opening configured to collect sample fluid and guide it into the microfluidic sample channel system. Specifically, the sample application port may correspond to a recess or orifice, specifically a tapered orifice and / or cutout in an element of the test carrier (e.g., a central element and / or a cover plate (first or second cover plate) of the test carrier) to allow entry into the microfluidic sample channel system (specifically, a microfluidic sample channel system located below the sample application port within a core element of the test carrier). In addition to the sample application port and sample release port, the microfluidic sample channel system may be provided with other openings and / or orifices to allow fluids, liquids, and / or gases to escape from the microfluidic sample channel system.

[0027] The test area includes a single, shared recessed measurement membrane region corresponding to a flat plane (substantially free of walls, bumps, and / or irregularities within the recessed measurement membrane region) defined at least partially by recessed walls (recessed edges), and is the area in which one or more measurement membranes can be located. In other words, the shared recessed measurement membrane region is a single region defined by recessed edges and simultaneously shared by several measurement membranes, such that sample fluid reaching the sample release port and released into the shared recessed measurement membrane region can be received substantially simultaneously by multiple measurement membranes. In other words, the shared recessed measurement membrane region does not include any walls except for the recessed edges that separate the measurement membranes from each other. The recessed region can be considered as a plane defined by the recessed edges forming the walls of the shared recessed measurement membrane region, and this plane is below the top surface of the cartridge, a portion of which is omitted and / or cut away to form the recess. The shared recessed measurement membrane region has a depth relative to the top surface of the cartridge, reflected by the depth of the recessed edges, and will be further described below in this specification. The fact that a shared recessed assay membrane area can be used by a single assay membrane or shared by two, three, four, or more assay membranes has the advantage that when more than one assay membrane is provided, no additional recessed assay membrane area needs to be added. In other words, the same cartridge used for a single test (providing only one assay membrane) can also be used for multiple tests (providing more than one assay membrane). Furthermore, only a single sample release port is required to access the shared recessed assay membrane area, and thus this single sample release port can be provided. Multiple assay membranes can be received by the shared recessed assay membrane area such that the distance d between two adjacent assay membranes is between approximately 0 mm and 5 mm, specifically between approximately 0.1 mm and 2 mm, and more specifically between approximately 0.3 mm and 1 mm. In other words, two adjacent assay membranes can be physically in contact along their length edges, or they can be very close to each other—they are not separated from each other and / or they are in different chambers and are not spatially separated. The chamber can be considered as a volume surrounded by walls for approximately 60-100% (specifically approximately 70-98%, and more specifically approximately 80-97%). Specifically, in one embodiment, there may be no wall between the two assay membranes. The test zone may include a waste pad region, which is an area in which one or more waste pads can be positioned. The test zone includes a sample release port for releasing at least one portion of the sample fluid into a shared recessed assay membrane region. The sample release port may be an opening that fluidly connects the microfluidic sample channel system to the test zone on the opposite side of the sample application port in the flow direction. In other words, fluid through the microfluidic sample channel system is guided to the test zone via the sample release port to control the release location of the sample fluid in the test zone. Compared to the microfluidic sample channel system, the test zone can provide a larger reservoir volume.

[0028] The test area may correspond to a region recessed substantially in the shape of a single test membrane and / or multiple test membranes oriented toward each other (i.e., side-by-side). In all cases, the test area may be configured to receive one and / or multiple test membranes. In other words, the test area provides space for positioning one and / or multiple test membranes therein. The test area may match the size of the test membrane or multiple test membranes at least in one dimension, such that the test membranes do not move and their positions relative to the test carrier are accurately defined. The test area may be optically accessible, such that a detector can detect test results provided on the test membrane, and / or an observer can read the results. The test area may be at least partially open, i.e., not covered by a cover or any other element of the test carrier. Alternatively, the test area may be covered by at least a partially transparent portion of a cover and / or another element of the test carrier. The recess may define a test area having a shared recessed test membrane area and a possible waste pad area. The shared recessed test membrane area may be recessed substantially in the shape of a single test membrane or multiple test membranes arranged side-by-side, and / or define the position in which the respective test membranes may be positioned. The shared recessed measurement membrane area may include one or more barriers (walls), each barrier (wall) located between two or more measurement membranes, thereby preventing liquid flow between the barriers and / or restricting movement of the measurement membranes to give the measurement membranes a predefined fixed position. The length of the recess, which may define and / or correspond to the test area, may be between approximately 0.5 mm and 300 mm, specifically between 1 mm and 100 mm. When the measurement membrane is positioned in or on the recess, the length of the recess may be measured along the overall flow direction of the measurement membrane, and the length of the recess may substantially correspond to the combined length of the waste pad and the measurement membrane, or substantially correspond to a length slightly smaller than the combined length of the waste pad and the measurement membrane (when overlap is expected), or substantially correspond to a length slightly larger than the combined length of the waste pad and the measurement membrane (when they do not overlap). The width of the recess, which may define and / or correspond to the test area, may be between approximately 0.05 mm and 50 mm, specifically between approximately 1 mm and 20 mm. The depth of the recess corresponding to the test area can be defined and / or between approximately 0.05 mm and 20 mm, specifically between approximately 1 mm and 15 mm, and this depth may or may not vary in different areas of the test area. If the depth is changed, the flow direction can be predefined, for example, from a shallower location to a deeper location.

[0029] Two or more measuring membranes may be positioned adjacent to each other, i.e., side-by-side within a shared recessed measuring membrane region. This adjacent positioning may refer to their longitudinal axes and / or overall flow directions being parallel. Alternatively, this adjacent positioning may refer to an angle between their longitudinal axes and / or overall flow directions during membrane assembly. The shape of the shared recessed measuring membrane region and / or the location of obstructions may define this orientation of the measuring membranes, which may be parallel or non-parallel, i.e., an angle between at least two measuring membranes.

[0030] The microfluidic sample channel system is configured to guide at least a portion of the sample fluid from the sample application port to the sample release port. The sample fluid can be passively delivered through at least a portion of the microfluidic sample channel system driven by capillary forces, van der Waals forces, and / or adhesive forces. Alternatively or additionally, the sample fluid can be passively delivered through at least a portion of the microfluidic sample channel system driven by an external force (specifically, by rotation of a cartridge / test carrier that generates centrifugal and centrifugal forces).

[0031] At least one portion of a sample fluid can refer to at least one component of the sample fluid and / or at least one portion of the sample fluid. For example, if whole blood corresponds to a sample fluid, then serum can be considered a component of whole blood. Simultaneously, serum is a part of whole blood. If whole blood is aliquoted within a microfluidic sample channel system, i.e., dispensed, then the aliquoted volume further delivered can be considered a part of whole blood. Therefore, the term at least one portion can be interpreted as "a portion" and / or "a component".

[0032] Microfluidic sample channel systems can deliver sample fluids in the µL range, such as capillary blood samples. Capillary blood samples are easily collected from the probe or foot, which is particularly convenient for newborns or patients who are typically unwilling to undergo routine procedures involving larger blood collections. Furthermore, collecting capillary blood samples from a patient can be performed rapidly compared to routine procedures that require larger blood collections.

[0033] A microfluidic sample channel system can be integrated into the test carrier by recessed traces or a tunnel-like system within the plastic core (test carrier core). The microfluidic sample channel system can be formed, for example, by two elements (such as two plates), one or both plates including recessed traces and forming the upper and lower walls of the microfluidic sample channel system, respectively. Thus, these two elements can at least partially surround the microfluidic sample channel system. The microfluidic sample channel system can be sealed by a sealing element sandwiched between the two plates. If the microfluidic sample channel system is integrated into the test carrier by a tunnel-like system and / or a tubular system within the plastic core (test carrier core), it can be essentially surrounded by only one part, thus eliminating the need for a sealing element. In other words, the test carrier core can have an integrated microfluidic sample channel system, and in this case, the test carrier core can represent and / or correspond to the entire test carrier, and no cover plate is required to close (enclose) the microfluidic sample channel system.

[0034] Tunnel-like and / or tubular systems may include portions having circular and / or elliptical cross-sections. Alternatively, the cross-section may have a different shape, such as polygonal, in at least a section and / or portion thereof. Microfluidic sample channel systems may constitute tunnel-like portions and / or functional elements with geometries different from typical tunnels. Microfluidic sample channel systems may integrate and / or connect to other elements not considered part of the microfluidic channel and / or microfluidic sample channel system, such as tubing, needles, pads, capillaries, and / or elements in the test carrier, which are either not microfluidic elements or are specific elements of the microfluidic sample channel system. Further, some elements integrated with the microfluidic sample channel system may be considered part of the microfluidic sample channel system but differ in shape from channels, tunnels, and / or tubing. Such elements may be fluidly connected to, or have been fluidly connected to, other elements of the microfluidic sample channel system, specifically to sample application ports and sample release ports. Further, such elements may have specific functions beyond simply guiding and / or (e.g., passively delivering fluid by capillary force), as further described below.

[0035] The first and / or second assay membranes may be configured (specifically, their sizes are suitably determined) to be positioned within a shared recessed assay membrane region, and / or may have been positioned and / or fixed within the shared recessed assay membrane region. The first assay membrane is configured to receive at least one portion of a first part of the sample fluid, i.e., a sub-part of a component and / or a portion of the sample fluid, such as a portion of serum extracted from a whole blood sample, which is released into the test area / region via a sample release port. The first assay membrane is configured to indicate at least one first analyte in the sample fluid (i.e., in at least one first part of the sample fluid).

[0036] The second assay membrane may be configured to be positioned within a shared recessed assay membrane region, and / or may already be positioned within the shared recessed assay membrane region, adjacent to the first assay membrane. The second assay membrane may be positioned parallel to the first assay membrane. Alternatively, the second assay membrane may be positioned at an angle relative to the first assay membrane. The flow initiation sides of the first and second assay membranes may be positioned close to the sample release port, such that the sample released via the sample release port first reaches the flow initiation side of each assay membrane. Therefore, the term "adjacent" may refer to two assay membranes arranged parallel to each other or having an angle between them, i.e., one assay membrane is inclined relative to the other.

[0037] Similar to the function of the first assay membrane, the second assay membrane is configured to receive at least a second portion of the sample fluid from the sample release port and indicate at least one second analyte in the second portion of the sample fluid and / or indicate at least one first analyte in the second portion of the sample fluid with a different sensitivity / measurement range than the first assay membrane. In other words, the second assay membrane can test the first analyte within a more specific sensitivity / measurement range for further quantification of the first analyte in the sample fluid, or the second assay membrane can test different analytes, i.e., second analytes. This allows for the provision of more information about the sample fluid, i.e., more test results on a single IVD assay system or on a single IVD assay system. Therefore, a single IVD assay system (multiplex assay) can be used, whereas previously two or more IVD assay systems (single assay) were used to obtain the same amount of information.

[0038] Therefore, the measuring membrane can be cut and / or shaped to a specific size, specifically the width suitable for the dimensions of a shared recessed measuring membrane region. The measuring membranes can be separated by gaps (i.e., air gaps). Alternatively, the measuring membranes can be in contact with each other without gaps between them. The measuring membranes can be disposed on a shared substrate (such as a sheet) such that their orientation relative to each other can be fixed, and / or the gaps between them are predefined on the substrate. The measuring membrane can specifically comprise individual sheets of cellulose, nitrocellulose, glass fiber, nylon, and / or cellulose-based materials and / or composed thereof. The function of the measuring membrane is based on capillary forces, which drag the liquid from the flow initiation side to the flow termination side. Thus, the liquid is passively transported through the respective measuring membrane. Different measuring membrane materials may result in different flow rates due to differences in capillary forces. All measuring membranes can be made of the same material, and therefore all measuring membranes can have the same flow rate. Alternatively, one or all measuring membranes can be made of a material different from that of another measuring membrane to achieve a specific flow rate for that measuring membrane that differs from the flow rates of the other measuring membranes.

[0039] The test carrier may already be equipped with a measurement membrane, such that the measurement membrane is already positioned in and / or on the test carrier. Alternatively, the measurement membrane may be provided separately and manually positioned in and / or on the test carrier by, for example, a user or supplier.

[0040] The test carrier may specifically correspond to a centrifugal test carrier, wherein the microfluidic sample channel system is configured to guide at least one portion of sample fluid in at least a segment / part of the microfluidic sample channel system (i.e., the portion between the sample application port and the sample release port) under the influence and / or control of centrifugal force.

[0041] Overall, IVD testing systems (which can correspond to microfluidic cartridges, specifically centrifuge cartridges) allow control over each step of a single assay, resulting in superior and laboratory-like assay performance, such as high sensitivity, high precision, and high accuracy.

[0042] The microfluidic sample channel system may have a total path / trace length of approximately 5 mm to 50 mm (specifically approximately 10 mm to 25 mm) between the sample application port and the sample release port. The microfluidic sample channel system may be configured to receive volumes of approximately 0.5 µL to 800 µL, specifically approximately 1 µL to 500 µL. The microfluidic sample channel system may include, but is not limited to, channel portions with a width and / or diameter of approximately 0.05 mm to 15 mm, specifically approximately 0.1 mm to 5 mm.

[0043] One or more assay membranes may be configured to contain, but are not limited to, a fluid volume of about 0.5 µL to 800 µL, specifically about 1 µL to 500 µL.

[0044] The recessed areas and / or elements that can form microfluidic sample channel systems or their components, and the microfluidic wash buffer channel systems or their components and / or test areas or their components may include one or more steps, one or more different depths, one or more ramps and / or circular and / or stepped surfaces to guide the flow of fluids and / or liquids.

[0045] Generally, providing one cartridge for two, three, or even more analytes is preferable to providing one cartridge for a single analyte, because the cartridge material used per assay can be reduced to about half, a third, or even less. This makes the present invention particularly environmentally friendly, as it greatly reduces waste.

[0046] The IVD testing system may further include a third assay membrane located in a shared recessed assay membrane region, adjacent to the first assay membrane and / or the second assay membrane, and configured to receive at least a third portion of the sample fluid from the sample release port and indicate at least one third analyte in the sample fluid (specifically in the third portion of the sample fluid) and / or indicate at least one first analyte in the sample fluid (specifically in the third portion of the sample fluid) with a sensitivity / measurement range different from that of the first assay membrane and / or the second assay membrane.

[0047] The third measuring membrane may be positioned parallel to the first and / or second measuring membranes. Alternatively, the third measuring membrane may be positioned at an angle relative to the first and / or second measuring membranes. The flow initiation sides of the first and / or second measuring membranes and the third measuring membrane may be positioned close to the sample release port, such that the sample released via the sample release port first reaches the flow initiation side. If more than two measuring membranes are provided, they may be arranged in a star shape, pointing towards a central point, specifically the center point where the sample release port is located and where the sample fluid is released.

[0048] The third assay membrane may have a function corresponding to the first and / or second membranes, i.e., detecting independent analytes or the same analytes detected by the first and / or second membranes, but with different sensitivity / measurement ranges. The IVD testing system may further include additional assay membranes with corresponding functions, i.e., four, five, six, seven, eight, nine, ten or more assay membranes.

[0049] The third measuring membrane may be separated from the first and / or second membranes and have gaps and / or may share a common substrate.

[0050] The first assay membrane may rely on a first capture mechanism configured to capture the first analyte after it has been labeled with a first capture reagent, and / or the second assay membrane may rely on a second capture mechanism different from the first capture mechanism to capture the second analyte after it has been labeled with a second capture reagent different from the first capture reagent, specifically when using a sandwich immunoassay (system) as the assay format. The first and / or second capture reagents may include one of a biotinylated antibody and / or a digoxigenin (dig)-labeled antibody. These antibodies may be configured to bind to the first and / or second analyte. The first and / or second capture mechanism may include one of: streptavidin for capturing the first or second analyte via a biotinylated antibody, anti-dig for capturing the first or second analyte via dig-labeled anti-dig, a DNA-based system, and / or an analyte-specific antibody for directly capturing the first or second analyte.

[0051] Choosing different capture mechanisms and corresponding reagents to label different analytes is advantageous, making the labeled analytes specific to the capture mechanism, or the capture mechanism specific to the labeling of the analytes, and allowing the labeled analytes to bind and / or be captured in spatially separated regions and / or lines. This enables differentiation between analytes and thus allows for multiple assays according to one embodiment. Such systems are typically based on sandwich immunoassays.

[0052] The first and second measuring membranes may each have an overall flow axis defined by a flow initiation side and a flow termination side opposite to the flow initiation side, the respective overall flow axis passing through the flow initiation side and the flow termination side, and wherein the overall flow axis of the first measuring membrane and the overall flow axis of the second measuring membrane may be aligned parallel to each other, such that the respective flow initiation side of each measuring membrane is positioned as the sample release port closest to the test area.

[0053] The overall flow axis of the first and / or second measuring membrane may correspond to the respective longitudinal axis of the measuring membrane. The overall flow axis may correspond to the direction in which the fluid flows substantially and / or is passively transported when it reaches the predetermined flow initiation side. Therefore, the fluid flows from the flow initiation side of the respective measuring membrane to and / or to the flow termination side of that measuring membrane. It should be understood that the fluid may also flow in another direction, but the overall flow axis and / or direction is (pre-defined) by the flow initiation side and the flow termination side, and the overall flow axis passes through both the flow initiation side and the flow termination side. The pre-defined flow initiation side may be the side closest to the line that the fluid should first pass through, such as the test (result) line.

[0054] The IVD testing system may further include at least one waste pad positioned to contact the first and / or second test membrane on the respective flow end side.

[0055] The waste pad can be configured and / or positioned to receive, at the respective flow end side, a fluid waste portion and / or washing buffer and / or washing solvent from the first and / or second assay membranes. The fluid waste portion corresponds to the portion of the sample fluid that passes through at least one assay membrane and reaches the waste pad, and the waste pad still has the volumetric capacity to contain the fluid waste portion.

[0056] The waste pad collects fluid or at least a portion of fluid that has passed through or via the measuring membrane. Therefore, the waste pad acts as a liquid reservoir, determining the flow rate through the measuring membrane once the liquid reaches it. Similar to the measuring membrane, the waste pad can function based on capillary forces, causing the liquid to be dragged by the capillary action of the waste pad. A first measuring membrane, a second measuring membrane, and possibly other measuring membranes can share a single waste pad; this design is easy to assemble and highly efficient in use. Alternatively, at least one measuring membrane can be connected to and / or contact a separate waste pad, configured to collect fluid only from that particular measuring membrane and not shared between measuring membranes. In one case, all the positioned measuring membranes are in contact with their respective waste pads, and the waste pads and measuring membranes are fluid-isolated, i.e., no liquid flows from one waste pad to another, nor from one measuring membrane to another. In this case, the flow rate can be set to an independent value and / or controlled not only by the measuring membrane itself but also by the waste pad. Once the liquid reaches the waste pad that comes into contact with the measuring membrane on the end of the flow, it will also affect the flow rate through the measuring membrane.

[0057] The waste pad may be positioned on the test carrier and at least partially within or beyond the test area, but in contact with the measurement membrane and / or fluid. The test carrier may include a specific waste pad region configured to receive one or more waste pads. The waste pad region may be substantially recessed in the shape of one or more waste pads. The waste pad region may have one or more barriers between the waste pads such that fluid flow between the barriers is stopped. The waste pad region may at least partially include the test area (specifically, a shared recessed measurement membrane region), or may be connected to and / or adjacent to a shared recessed measurement membrane region.

[0058] The test carrier may already be equipped with a waste pad, so that the waste pad is already positioned in and / or on the test carrier. Alternatively, the waste pad may be provided separately so that it can be positioned in and / or on the test carrier, for example, by the user (or by the supplier).

[0059] The length of one or more waste pads may be, but is not limited to, about 0.5 mm to 100 mm, specifically about 1 mm to 70 mm, and more specifically about 5 mm to 50 mm. The width of one or more waste pads may be, but is not limited to, about 0.01 mm to 50 mm, specifically about 0.5 mm to 30 mm, and more specifically about 1 mm to 20 mm. The thickness of one or more waste pads may be, but is not limited to, about 0.05 mm to 20 mm, specifically about 0.08 mm to 15 mm, and more specifically about 0.1 mm to 10 mm. One or more waste pads may be, but is not limited to, at least one of the following materials: cotton, cellulose, or synthetic fibers (such as polypropylene). One or more waste pads may be configured to receive, but is not limited to, a fluid volume of about 0.5 µL to 700 µL, specifically about 1 µL to 500 µL.

[0060] IVD testing systems may further include coupling pads that can be configured to provide couplings and separate components from the sample fluid and / or another pad with a different function.

[0061] The test carrier may include a plasma separation element integrated with a microfluidic sample channel system and configured to extract plasma from the sample fluid when the sample fluid includes whole blood. The microfluidic sample channel system may be configured to direct at least one portion of the sample fluid from the sample application port to the plasma separation element and to direct at least one portion of the plasma from the plasma separation element to the sample release port.

[0062] Plasma separation elements may correspond to elements of microfluidic sample channels, differing in shape from pipes, channels, and / or fittings, and having the function of separating plasma from whole blood samples. If the test carrier corresponds to a centrifugal test carrier, the plasma separation element may be configured to separate plasma from whole blood samples under the influence and / or control of centrifugal force. Alternatively or additionally, plasma separation elements may correspond to or include elements (such as plasma separation pads and / or coupling pads with separation functions) configured to separate plasma from whole blood as a whole blood sample passes through the element, the coupling pad including a membrane that retains components other than plasma from the whole blood. Particularly for immunoassays, it may be necessary to separate plasma from whole blood for further processing (e.g., labeling) and sample analysis.

[0063] The integration of a plasma separation element with a microfluidic sample channel system may mean that the plasma separation element is a component of the microfluidic sample channel system, and / or that the plasma separation element is fluidly connected to or can be connected to a portion of the microfluidic sample channel system. In other words, the plasma separation element may be connected to and / or can be connected to a portion of the microfluidic sample channel system, and / or may correspond to a portion and / or component of the microfluidic sample channel system in the form of a chamber, hollow geometry, and / or channel. A portion of the microfluidic sample channel system may passively deliver sample fluid to the plasma separation element via capillary forces, while the plasma separation element may perform its function based on other forces, such as centrifugal force, but is not limited to these. However, if, for example, a plasma separation pad and / or a coupling pad with separation function are involved, the function of the plasma separation element may also be based on capillary forces.

[0064] The plasma separation element may have a volume of, but is not limited to, about 0.5 µL to 700 µL, specifically about 1 µL to 500 µL, and more specifically about 5 µL to 300 µL. The plasma separation element may be configured to extract plasma from a sample fluid in a volume of, but not limited to, about 0.5 µL to 400 µL, specifically about 0.8 µL to 300 µL, and more specifically about 1 µL to 200 µL.

[0065] The test carrier may further include at least one reagent chamber integrated with a microfluidic sample channel system and used to contain at least one reagent configured to bind with at least one analyte. The at least one reagent chamber may include: at least one capture reagent chamber for containing a first capture reagent and / or a second capture reagent. The at least one reagent chamber may already contain and / or contain at least one reagent, specifically a first capture reagent and / or a second capture reagent. The microfluidic sample channel system may be configured, for example, to guide at least a portion of the sample fluid from a sample application port to the at least one reagent chamber by capillary force, and to guide at least a portion of the sample fluid within the at least one reagent chamber to a release port.

[0066] At least a portion of the sample fluid may be contacted, mixed, and / or incubated with at least one reagent, such that at least one target analyte can be labeled with a capture reagent designed to specifically bind to the analyte's binding site. At least one capture reagent chamber may be specifically configured to contain and / or contain more than one capture reagent. For example, a capture reagent chamber may contain two capture reagents to label two target analytes in the sample fluid, respectively. Capture reagent chambers may contain even more than two capture reagents, such as three, four, five, six, seven, nine, ten, or more capture reagents. The capture reagent may be provided in dry form and, when guided through the capture reagent chamber, may only be dissolved by the sample fluid and / or a portion (a portion and / or component) of it.

[0067] If the test carrier is a centrifugal test carrier, the sample fluid can be guided into at least one capture reagent chamber under the control of centrifugal force; however, if the passive delivery of fluid to at least one capture reagent chamber is based on capillary force, centrifugal force may not be required, for example, when the sample fluid is delivered from a plasma separation element to at least one capture reagent chamber. More specifically, at least a portion of the sample fluid can be guided into the capture reagent chamber under the control of centrifugal force via a microfluidic sample channel system and possibly via other elements (such as a plasma separation element). Further, after at least a portion of the sample fluid can be incubated in the capture reagent chamber for a period of time, at least a portion of the sample fluid in the capture reagent chamber can be guided to the sample release port by means of capillary force and / or centrifugal force via another portion and / or other elements of the microfluidic channel system. With regard to the capture reagent chamber, the term "integrated with" can also refer to components of the microfluidic channel system and / or components connected to at least a portion of the microfluidic channel system; specifically, the capture reagent chamber may not have the function of passively delivering sample fluid by means of capillary force, while other elements and / or portions of the microfluidic channel system may have the function of passively delivering sample fluid.

[0068] The volume of one or more reagent chambers may be, but is not limited to, from about 0.5 µL to 700 µL, specifically from about 1 µL to 500 µL. One or more reagent chambers may be configured, but is not limited to, to receive and / or contain reagents with a mass of about 1 µg to 100 mg, specifically from about 5 µg to 50 mg.

[0069] At least one reagent chamber may include at least one detection reagent chamber for containing at least one first detection reagent and / or for containing a second capture reagent, the first detection reagent being configured to bind to a first analyte and the second capture reagent being configured to bind to a second analyte.

[0070] At least a portion of the sample fluid may be contacted, mixed, and / or incubated with at least one detection reagent, such that at least one target analyte can be labeled with the detection reagent, which is designed to specifically bind to the (other) binding site of the analyte. At least one detection reagent chamber may be specifically configured to contain and / or contain more than one detection reagent. For example, a detection reagent chamber may contain two detection reagents to label two target analytes in the sample fluid, respectively. Detection reagent chambers may contain even more than two detection reagents, such as three, four, five, six, seven, nine, ten, or more capture reagents. Detection reagents may be provided in dry form and, when guided through a detection reagent chamber, may only be dissolved by the sample fluid and / or a portion (a portion and / or component) of it.

[0071] If the test carrier is a centrifugal test carrier, the sample fluid can be guided into at least one test reagent chamber under the control of centrifugal force; however, if the passive transport of fluid to at least one test reagent chamber is based on capillary force, centrifugal force may not be required, for example, when the sample fluid is transported from the capture reagent chamber to at least one test reagent chamber. More specifically, at least a portion of the sample fluid can be guided into the test reagent chamber via a microfluidic sample channel system and possibly via other elements (such as plasma separation elements and / or capture reagent chambers) under the control of centrifugal force and / or capillary force. Further, after at least a portion of the sample can be incubated in the test reagent chamber for a period of time, at least a portion of the sample fluid in the test reagent chamber can be guided to the sample release port by means of capillary force and / or centrifugal force via another portion of the microfluidic channel system and / or other elements. With regard to the reagent chamber, the term "integrated into" may also refer to components of the microfluidic channel system and / or components connected to at least a portion / part of the microfluidic channel system; specifically, the reagent chamber may not have the function of passively delivering sample fluid by means of capillary force, while other elements and / or portions of the microfluidic channel system may have the function of passively delivering sample fluid by means of capillary force.

[0072] The test carrier may already be equipped with reagents (capture reagents and / or detection reagents), so that the reagents are already present in and / or on the test carrier. Alternatively, the reagents may be provided by the user (e.g., or by the supplier) to be positioned in and / or on the test carrier.

[0073] It is advantageous to have dedicated reagent chambers for mixing and incubating the sample fluid with the reagent, as the incubation period in each chamber can be controlled individually, for example, by slowing the centrifuge speed and / or by stopping the centrifuge rotation and / or by allowing a portion of the sample fluid to settle within the reagent chamber. Alternatively or additionally, the reagent can be placed in or on a pad (such as a coupling pad) through which at least a portion of the sample fluid can pass—in which case the incubation period can be controlled by the flow rate through the pad. Coupling pads can be placed between sections of the microfluidic channel system and / or within the test area. Furthermore, the reagent can be provided as a dry powder or in solution form. Dry forms (such as powders) are long-term stable.

[0074] If only two measuring membranes are provided in the test assembly, the width of each of the first and second measuring membranes may be, but is not limited to, about 1.5 mm to 2 mm, and may be measured in a direction perpendicular to their respective overall flow axis (e.g., parallel to one or more lines, such as test lines); or if three measuring membranes are provided in the test assembly, the width of each of the first, second, and third measuring membranes may be, but is not limited to, about 1 mm to 1.3 mm, and may be measured in a direction perpendicular to their respective overall flow axis. When a single membrane is used to test a single analyte, the width may vary between, but is not limited to, about 3 mm and 4 mm. The width of the measuring membrane is typically measured perpendicular to the overall flow direction of the measuring membrane. The more measuring membranes provided, the smaller the width must be selected to arrange all measuring membranes adjacent to each other in the test area. The length of one or more measuring membranes may be about 5 mm to 25 mm, specifically about 10 mm to 20 mm, and more specifically about 15 mm to 18 mm. The length of the measuring membrane is typically measured along the overall flow direction and / or perpendicular to lines such as test lines. The thickness of one or more measuring membranes may be, but is not limited to, about 0.1 mm to 10 mm. One or more measuring membranes may include at least one of the following materials: cellulose, nitrocellulose, glass fiber, nylon and / or cellulose-based materials.

[0075] The test area may include a wash buffer release port, and the test carrier may further include, but is not limited to: a microfluidic wash buffer channel system for guiding and / or passively delivering the wash buffer, for example, based on capillary force and / or centrifugal force; and a wash buffer reservoir receiver for accommodating and / or receiving a wash buffer reservoir. The test carrier may include a fluidly connected wash buffer reservoir, or may be configured to be fluidly connected to the wash buffer release port via the microfluidic wash buffer channel system to provide a predetermined amount of wash buffer to the test membrane at the corresponding flow initiation side.

[0076] A wash buffer reservoir may correspond to a wash buffer blister, configured to be fluidly connected to a wash buffer release port via a microfluidic wash buffer channel system to provide a predetermined amount of wash buffer to the assay membrane on the corresponding flow initiation side, for example, under the influence / control of centrifugal force. The fluid connection can be established by puncturing the wash buffer blister with a sharp element and / or a needle. This can be performed in an automated manner.

[0077] Blisters may include, but are not limited to, at least one of the following materials: aluminum, PVC (polyvinyl chloride), PET (polyethylene terephthalate), and PVDC (polyvinylidene chloride).

[0078] The wash buffer reservoir receiving section may correspond to the recess in the test carrier for accommodating and / or receiving the wash buffer reservoir.

[0079] The wash buffer system allows for the provision of a wash buffer to the assay membrane to remove non-specifically adsorbed and / or weakly bound molecules and / or substances from the assay membrane, thereby obtaining accurate test results that simultaneously provide high sensitivity and selectivity.

[0080] The test vehicle may already be equipped with a wash buffer reservoir, such that the wash buffer reservoir, and in particular the wash buffer, is already present in and / or on the test vehicle. Alternatively, a wash buffer reservoir may be provided so that it can be positioned in and / or on the test vehicle by the user (e.g., or by the supplier).

[0081] The volume of the wash buffer reservoir may be, but is not limited to, approximately 10 µL to 5 mL, specifically approximately 100 µL to 3 mL. The microfluidic wash buffer channel system may, but is not limited to, have a total path / trace length of approximately 1 mm to 20 cm (specifically approximately 5 mm to 7 cm) between the opening of the wash buffer reservoir or the starting point of the microfluidic wash buffer channel system and the wash buffer release port. The microfluidic wash buffer channel system may be configured to receive volumes of approximately 1 µL to 5 mL, specifically approximately 50 µL to 2 mL. The microfluidic wash buffer channel system may include, but is not limited to, channel portions with a width and / or diameter of approximately 0.5 mm to 75 mm. The wash buffer reservoir receiving portion may be, but is not limited to, circular, elliptical, polygonal, and / or conical. The diameter or length and / or width of the wash buffer reservoir receiving portion may be, but is not limited to, approximately 0.1 µm to 2 cm. The wash buffer reservoir receiving portion may have a ring-shaped retaining frame. The wash buffer reservoir may be, but is not limited to, circular, elliptical, polygonal, and / or conical. The wash buffer reservoir may have a cross-section with a diameter or length and / or width of approximately 0.1 µm to 2 cm (specifically approximately 5 µm to 1 cm).

[0082] The first and / or second analytes may include, but are not limited to, cardiac parameters, specifically at least one of the cardiac analytes, such as cTropT, NTproBNP, D-dimer, PCT, BM2, BM3, creatinine, potassium, and sodium chloride. Alternatively or additionally, other analytes and / or parameters besides those listed above can be detected by an IVD testing system. The listed analytes provide information about a patient's cardiac health. Specifically, one or more of these parameters can be used to identify myocardial infarction / coronary thrombosis and / or other cardiac conditions. IVD testing systems allow for multiplex detection and / or better quantification of one or more of these parameters, enabling physicians and / or healthcare professionals to better and more effectively identify diseases and / or acute health conditions threatening patients, such as heart disease. Because IVD testing systems can provide multiple test results in a short period of time, the likelihood of ensuring patient safety is greatly increased.

[0083] Multiple diagnostic assays may correspond to immunochromatographic assays, and at least one of the first detection reagents and / or at least one second detection reagent may include a fluorescent molecule. Immunochromatographic assays typically provide reliable, sensitive, and / or selective test results. Therefore, an assay membrane may correspond to an immunochromatographic assay membrane.

[0084] The test carrier may correspond to a centrifugal microfluidic test carrier, and at least a segment and / or portion of the microfluidic channel system, comprising a microfluidic sample channel system and a microfluidic wash buffer system, may be configured to, under the influence of centrifugal force, guide at least a portion of the sample fluid from the sample application port to the sample release port, and / or guide at least a portion of the wash buffer from the blister to the wash buffer release port. Flow can be controlled by selecting a specific rotation frequency and / or stopping rotation, through the corresponding centrifugal force during rotation and / or the capillary force when rotation is stopped. Alternatively or additionally, in at least a segment and / or portion of the microfluidic sample channel system, capillary force may drive the delivery of the sample fluid.

[0085] To guide a portion of the sample fluid through the entire microfluidic sample channel system, a program for a predetermined rotation frequency can be run / executed. This program may include quantitatively accelerating to a certain value to move the sample fluid, decelerating to reduce sample fluid movement, stopping to allow the sample fluid to settle in one location (e.g., for incubation), and / or repeatedly accelerating to further move the sample fluid toward the sample release port. Therefore, the centrifugal microfluidic test carrier is configured to, under the influence of and / or control of centrifugal force, guide at least a portion, at least partially, of the sample fluid from the sample application port to the sample release port.

[0086] According to a second aspect of the invention, an IVD testing apparatus includes: a centrifuge having a cartridge support plate configured to rotate about a centrifugal axis via a motor of the centrifuge; a controller configured to control the rotation of the motor of the centrifuge and the cartridge support plate; and at least one cartridge comprising and / or corresponding to an IVD testing system including a centrifuged microfluidic test carrier, wherein the cartridge is configured to be fixed to the cartridge support plate so as to rotate together with the cartridge support plate rotating about the centrifugal axis.

[0087] The IVD testing apparatus provides all the advantages and technical effects already described for an IVD testing system included in the IVD testing apparatus or any corresponding embodiment thereof. The IVD testing system may be provided to the IVD testing apparatus in the form of a cartridge. The cartridge may correspond to the IVD testing system, or may include the IVD testing system described according to any of the embodiments described in the examples.

[0088] The cartridge support plate can be configured to receive two or more IVD testing systems located off-axis, with the centrifugal axis passing perpendicularly through the cartridge support plate when the cartridge support plate is properly positioned within the IVD testing device. Preferably, the two or more IVD testing systems are symmetrically positioned around the centrifugal axis.

[0089] According to a second aspect of the invention, a method for performing multiple diagnostic assays includes the following steps: providing an IVD testing apparatus as described above; positioning and / or securing at least one cartridge to a cartridge support plate; initiating the delivery of at least one portion of sample fluid through a microfluidic sample channel system to a first assay membrane and a second assay membrane, such that at least one portion can be delivered through the first assay membrane and the second assay membrane; detecting whether the first assay membrane indicates the presence of a first analyte and / or whether the second assay membrane indicates the presence of a second analyte.

[0090] The method for performing multiple diagnostic assays offers all the advantages and technical effects already described for IVD testing systems or any embodiment thereof.

[0091] Initiating the delivery of at least one portion of the sample fluid through the microfluidic sample channel system may include adding the sample fluid to or onto the sample application port. Passive delivery can then be achieved by means of capillary forces generated by the walls of the microfluidic sample channel system. When a cartridge support plate with a cartridge is rotated about a rotation axis in a centrifuge at at least a first predetermined rotational frequency, the delivery of a portion of the sample fluid in at least one portion of the microfluidic sample channel system may be additionally supported by centrifugal force. The first predetermined rotational frequency is configured to guide and / or deliver at least one portion of the sample fluid through the microfluidic sample channel system in at least a segment between the sample application port and the sample release port, to efficiently and / or rapidly provide at least one portion of the sample fluid to a first assay membrane, a second assay membrane, and possibly a third assay membrane, enabling efficient execution of multiplex diagnostic assays.

[0092] Rotating a cartridge support plate with a cartridge about a rotation axis in a centrifuge at at least one first predetermined rotational frequency may include a procedure of accelerating, decelerating, and stopping rotation, wherein the first predetermined rotational frequency is configured to guide, move, and / or deliver at least one portion of the sample fluid from the sample application port through a microfluidic sample channel system to the sample release port. For example, the procedure may include at least one of the following method steps:

[0093] The method may include the step of initiating rotation by accelerating a cartridge support plate with a cartridge at at least one first predetermined rotational frequency ω1 about a rotational axis in a centrifuge, the first predetermined rotational frequency ω1 being configured to guide at least one portion of the sample fluid from the sample application port through a first portion of a microfluidic sample channel system to a plasma separation element. The first predetermined rotational frequency ω1 may be between about 0 Hz and 100 Hz, specifically between about 55 Hz and 85 Hz, more specifically about 70 Hz. If delivery relies solely on capillary force, the first predetermined rotational frequency ω1 may be equal to zero.

[0094] The method may include the step of rotating a cartridge support plate at at least one second predetermined rotation frequency ω2, the at least one second predetermined rotation frequency ω2 being configured to separate / extract plasma from whole blood. The second predetermined rotation frequency ω2 may be between about 50 Hz and 115 Hz, specifically between about 70 Hz and 100 Hz, and in some embodiments, the second predetermined rotation frequency ω2 may exceed 100 Hz or even exceed 115 Hz.

[0095] The method may include the step of rotating a cartridge support plate at at least one third predetermined rotational frequency ω3, the third predetermined rotational frequency ω3 being configured to guide, move, and / or deliver at least one portion of plasma from a plasma separation element via a second portion of a microfluidic sample channel system to a capture reagent chamber. The third predetermined rotational frequency ω3 may be between about 0 Hz and 100 Hz, specifically between about 55 Hz and 85 Hz, more specifically about 70 Hz. If delivery relies solely on capillary force, the third predetermined rotational frequency ω3 may be equal to zero.

[0096] The method may include steps including stopping rotation and continuing a predetermined first cycle—if not already stopped—the predetermined first cycle being configured to allow plasma to dissolve, mix, and / or incubate with capture reagents in a capture reagent chamber, such that at least one capture reagent can label a predetermined analyte via specific binding to a first binding site of the predetermined analyte.

[0097] The method may include the step of rotating a cartridge support plate at at least one fourth predetermined rotational frequency ω4, the fourth predetermined rotational frequency ω4 being configured to guide, move, and / or deliver at least one portion of plasma carrying a labeled analyte from a capture reagent chamber to a detection reagent chamber via a third portion of a microfluidic sample channel system. The fourth predetermined rotational frequency ω4 may be between about 0 Hz and 100 Hz, specifically between about 55 Hz and 85 Hz, more specifically about 70 Hz. If delivery relies solely on capillary force, the fourth predetermined rotational frequency ω4 may be equal to zero.

[0098] The method may include steps including stopping rotation and continuing a predetermined second cycle—if not already stopped—that is, the predetermined second cycle being configured to allow plasma (which has an analyte labeled with a capture reagent) to dissolve, mix and / or incubate with the capture reagent in the detection reagent chamber, such that at least one detection reagent can label the predetermined analyte via specific binding to a second binding site of the predetermined analyte.

[0099] The method may include the step of rotating a cartridge support plate at at least one fifth predetermined rotational frequency ω5, the at least one fifth predetermined rotational frequency ω5 being configured to guide, move, and / or deliver at least one portion of plasma carrying a double-labeled / sandwich-labeled analyte from the test reagent chamber via a fourth portion of a microfluidic sample channel system to a sample release port to provide at least one portion of plasma carrying the sandwich-labeled analyte to a first assay membrane or a second assay membrane. The fifth predetermined rotational frequency ω5 may be between about 0 Hz and 100 Hz, specifically between about 55 Hz and 85 Hz, more specifically about 70 Hz. If delivery relies solely on capillary force, the fifth predetermined rotational frequency ω5 may be equal to zero.

[0100] The method may include steps including stopping rotation and continuing a predetermined third cycle—if not already stopped—the predetermined third cycle being configured to allow multiple assays to be performed by allowing at least a portion of plasma having sandwich-marked analytes to flow along the respective overall flow axes of the first and second assay membranes from the flow initiation side toward the flow termination side and at least one waste pad.

[0101] The method may include the step of puncturing a wash buffer reservoir to establish a fluid connection between the wash buffer reservoir and a microfluidic wash buffer channel system.

[0102] The method may include the step of rotating a cartridge support plate at at least one sixth predetermined rotational frequency ω6, the sixth predetermined rotational frequency ω6 being configured to guide, move, and / or deliver at least one portion of the wash buffer from a wash buffer reservoir via a first portion of a microfluidic wash buffer channel system to a wash buffer dispensing element and / or chamber. The sixth predetermined rotational frequency ω6 may be between about 0 Hz and 100 Hz, specifically between about 55 Hz and 85 Hz, more specifically about 70 Hz. If delivery relies solely on capillary force, the sixth predetermined rotational frequency ω6 may be equal to zero.

[0103] The method may include the step of rotating a cartridge support plate at at least one seventh predetermined rotation frequency ω7, the seventh predetermined rotation frequency ω7 being configured to aliquot / dispense the total amount of washing buffer in the washing buffer aliquoting element and / or chamber to obtain at least one predetermined portion of the washing buffer. The seventh predetermined rotation frequency ω7 may be between about 30 Hz and 70 Hz, specifically between about 40 Hz and 60 Hz, such as 50 Hz; in some embodiments, the seventh predetermined rotation frequency ω7 may exceed 50 Hz or even exceed 70 Hz.

[0104] The method may include the step of rotating a cartridge support plate at at least one eighth predetermined rotational frequency ω8, the eighth predetermined rotational frequency ω8 being configured to guide, move, and / or deliver at least a portion of a wash buffer from a wash buffer dispensing element and / or chamber via a second portion of a microfluidic wash buffer channel system to a wash buffer release port at a sample area, to provide at least a portion of the at least one portion of the wash buffer to a first assay membrane and a second assay membrane. The eighth predetermined rotational frequency ω8 may be between about 0 Hz and 100 Hz, specifically between about 55 Hz and 85 Hz, more specifically about 70 Hz. If delivery relies solely on capillary force, the eighth predetermined rotational frequency ω8 may be equal to zero.

[0105] The method may include steps comprising stopping rotation and continuing for a predetermined fourth cycle—if not already stopped—the predetermined fourth cycle being configured to allow a washing step to be performed by allowing at least a portion of at least one equal portion of the washing buffer to flow along the respective overall flow axes of the first and second test membranes from the flow initiation side toward the flow termination side and at least one waste pad. Upon completion of the procedure, test results can be read from the test membranes by eye and / or camera and / or other detection devices.

[0106] The wash buffer aliquot element can be configured to receive, but is not limited to, amounts between about 0.05 µL and 150 µL, specifically between about 1 µL and 100 µL. The wash buffer aliquot element can be configured to divide the buffer into two, three, four, or five portions, each with a volume of, but not limited to, about 0.05 µL and 150 µL, specifically about 1 µL and 100 µL.

[0107] It goes without saying that regardless of whether the actual sample fluid contains the target analyte, the characteristics associated with labeling and / or detecting the target analyte are achieved. Multiple determinations are used to determine the presence of one or two or more analytes, and therefore, samples that do not contain a specific one or two or more analytes can also be tested. Detailed Implementation

[0108] In the following description, some exemplary embodiments will be described in detail, wherein the invention should not be construed as being limited to the described exemplary embodiments. The following examples and drawings are provided to aid in understanding the invention, and the true scope of the invention is set forth in the appended claims. Individual features described in particular embodiments may be combined arbitrarily, as long as they are not mutually exclusive. Furthermore, the different features provided together in the exemplary embodiments should not be considered as limiting the invention.

[0109] Those skilled in the art will understand that the elements in the accompanying drawings are shown for simplicity and clarity and are not necessarily drawn to scale. For example, the dimensions of some elements may be enlarged relative to other elements, while other elements may have been omitted or represented by reduced quantities to enhance clarity and improve understanding of aspects of this disclosure. The same reference numerals are used for the same or similar elements or elements having similar or identical effects in different embodiments and examples. Attached Figure Description

[0110] Figure 1a is a schematic diagram of the preparation of sandwich-labeled analytes according to known techniques;

[0111] Figure 1b is a schematic diagram of the detection of sandwich-labeled analytes on an immunochromatographic assay membrane according to known techniques;

[0112] Figure 2 is a top perspective view of an example IVD testing system, which includes an immunochromatographic assay membrane disposed in a centrifuged microfluidic test carrier.

[0113] Figure 3a is a schematic top perspective view of an IVD testing system according to the example in Figure 2, which includes an immunochromatographic assay membrane disposed within a centrifugal microfluidic test carrier for detecting analytes on an assay membrane.

[0114] Figure 3b is a schematic top perspective view of an IVD testing system according to one embodiment, which includes two immunochromatographic assay membranes disposed in a centrifuged microfluidic test carrier for multiplex detection of analytes;

[0115] Figure 4 is an exploded top perspective view of an IVD testing system according to another embodiment, which includes three immunochromatographic assay membranes disposed in a centrifuged microfluidic test carrier for multiplex detection of analytes;

[0116] Figure 5a is a schematic top perspective view of an IVD testing system according to an embodiment of Figure 3b;

[0117] Figure 5b is a schematic diagram of the first capture mechanism of the first assay membrane, which is set as the immunochromatographic assay element of Figure 5a.

[0118] Figure 5c is a schematic diagram of the second capture mechanism of the second assay membrane, which is set as the immunochromatographic assay element of Figure 5a;

[0119] Figure 6a is a front view schematic diagram of an IVD testing apparatus including an IVD testing system according to an embodiment;

[0120] Figure 6b is a top view of a cartridge support plate of three cartridges with the IVD testing apparatus of Figure 6a according to an embodiment;

[0121] Figure 7 is a schematic flowchart of a method for performing multiple diagnostic measurements according to one embodiment; and

[0122] Figures 8a-8h are schematic diagrams of the cutting lines A-A', a-a', BB' and bb' indicated in Figures 3a and 3b according to different embodiments.

[0123] Figure 1a is a schematic diagram illustrating a method for preparing sandwich-labeled analyte 31c (an analyte labeled with two reagents at two different binding sites) according to known techniques, and Figure 1b is a schematic diagram illustrating a method for detecting sandwich-labeled analyte 31c on an immunochromatographic assay membrane 6 according to known techniques. The methods related to Figures 1a and 1b are further described in the background section above.

[0124] Typically, when using immunochromatography for multiplex detection in immunoassays—that is, simultaneously identifying multiple analytes from the same sample in a single measurement—assay membranes with more than one test line are used. Each test line is coated with an analyte-specific antibody for specifically capturing the target analyte. However, conventional assays include assay interference: in kit formats, each of the test lines with corresponding fixed reagents is a source of nonspecific interactions between the assay reagents and the analytes, as these reagents must flow along the other test lines. Nonspecific interactions between the analyte-specific detection antibody and other reagents flowing along the assay membrane, as well as with the reagents in the capture zone, are well known in the field. Therefore, this invention improves accuracy and signal-to-noise ratio, which may arise from dependence on signal recovery rates between donors.

[0125] Furthermore, this invention can improve or even optimize the distance between test leads to reduce and / or prevent crosstalk between test leads during signal detection. Therefore, it eliminates the need for longer membranes, which might otherwise be required; and avoids the need to redesign the test carrier to provide more space for the membrane. Therefore, the advantage of this invention is that test kits for single-parameter testing and test kits for multiple detection can be used with the same platform, hardware, equipment, and / or test carrier. Therefore, different cartridge designs are not required. No additional design of the test carrier is needed in the production line, thus avoiding the complexity of material management and production.

[0126] It may also be unnecessary to design test carriers with separation channels (including assay membranes) for each of the multiple analytes behind the public blood application area. Therefore, the size and complexity of the test carriers do not need to be changed and / or undergo an increase in complexity, which could lead to increased production costs. Furthermore, the increased sample volume required to run multiplex assays can be avoided.

[0127] Therefore, an advantage of this invention is that it provides a platform for both single and multiplex immunochromatographic assays. This platform offers the flexibility to perform single or multiple assays on the same point-of-care (POC) platform without requiring significant changes to the design of the assay carriers and instruments, or the required sample volumes.

[0128] Terminology parameters specifically include the analyte. Typically, terminology parameters may also include the properties of the sample fluid, hematocrit (Hct) value, coagulation status, etc.

[0129] Figure 2 is a schematic diagram of an example IVD testing system 1c, which includes a single immunochromatographic assay membrane 6 located in or on a centrifugal microfluidic test carrier 2. In embodiments of the invention, i.e., in IVD testing systems 1a and 1b of Figures 3b, 4, and 5a, all elements except the single assay membrane 6 can be used. In other words, the elements of the centrifugal microfluidic test carrier 2 and the waste pad 9 can be the same as those in embodiments of the invention. Similarly, in other words, by replacing the single assay membrane 6 with two or three or more assay membranes 6, 7, 8 that are narrower than the single assay membrane shown in Figure 2, the IVD testing system 1c shown in Figure 2 can be converted into one of the IVD testing systems 1a and 1b according to embodiments of the invention. Therefore, embodiments of the invention can be implemented using all the features described for the example of Figure 2 that are not directly related to the assay membrane 6. The IVD testing system 1a of Figure 3b is for multiplex detection and provides two assay membranes 6, 7. The IVD testing system 1b in Figure 4 is used for multiple detection and provides three assay membranes 6, 7, and 8.

[0130] The assay membrane 6 can be positioned, specifically loosely positioned, clamped, snapped, adhered, and / or fixed to the centrifugal microfluidic test carrier 2. The centrifugal microfluidic test carrier 2 is shown without one or both cover plates 2a, 2b and without a wash buffer reservoir 13 (e.g., as shown in FIG. 4). FIG. 4 relates to an embodiment of the invention based on multiple detection of three assay membranes 6, 7, 8, with the center / core 2c shown together with the first cover plate 2a and the second cover plate 2b, wherein the first cover plate 2a is configured to cover a first side S1 of the center / core 2c, and / or the size of the first cover plate is determined to cover the first side of the center / core, and / or the first cover plate is configured to match to cover the first side of the center / core; and the second cover plate 2b is configured to cover a second side S2 of the center / core 2c, and / or the size of the second cover plate is determined to cover the second side of the center / core, and / or the second cover plate is configured to match to cover the second side of the center / core. Therefore, the center component / core component 2c is sandwiched between the first cover plate 2a and the second cover plate 2b. It can be imagined that the center component / core component 2c is covered by only a single cover plate 2a or 2b, and thus the center component / core component 2c can be considered as either the upper or lower test carrier component. In Figure 2, for simplicity, only the center component and / or core component 2c of the test carrier 2 are shown, which can correspond to injection-molded elements. Therefore, the microfluidic test carrier 2 (see...) Figure 4 It may include one or both of the center / core 2c and the first cover plate 2a and the second cover plate 2b.

[0131] The microfluidic test carrier 2, together with the cover plates 2a, 2b, the test membrane 6, the waste pad 9, and the wash buffer reservoir 13 (not shown here), can be considered as a functional cartridge 24 that can be used in the IVD test apparatus 20, as schematically shown in Figure 6a.

[0132] The functional elements of the IVD testing system 1c (specifically the microfluidic test carrier 2) are described below as embodiments explicitly applicable to the present invention. A sample application port 3 is disposed on at least one element of the microfluidic test carrier 2. The sample application port 3 can be considered as the starting point and / or inlet for sample fluid 30 (not shown here) to enter the microfluidic sample channel system 5, which in this example may be, for example, whole blood. The microfluidic test carrier 2 is particularly suitable for testing analytes that may be present in whole blood samples. However, it is conceivable that another type of sample fluid should be tested, and the test carrier may be suitable for testing analytes that may be present in sample fluids such as urine, saliva, or other samples. Covers 2a, 2b may have recesses, cuts, and / or holes at the location of the sample application port 3 to provide access and / or inlet to the microfluidic sample channel system 5. A first portion of the microfluidic sample channel system 5 connects the sample application port 3 to the plasma separation element 10. In fact, the sample application port 3 and the plasma separation element 10 can be considered as segments, parts and / or elements of the microfluidic sample channel system 5, and the first part of the microfluidic sample channel system 5 connecting the sample application port 3 and the plasma separation element 10 can be considered as the first connection part of the microfluidic sample channel system 5.

[0133] A second portion of the microfluidic sample channel system 5 connects the plasma separation element 10 to the first reagent chamber 11a. A third portion of the microfluidic sample channel system 5 connects the first reagent chamber 11a to the second reagent chamber 11b. In fact, reagent chambers 11a and 11b can be considered segments, parts, and / or elements of the microfluidic sample channel system 5, and the second and third portions of the microfluidic sample channel system 5 can be considered the second and third connecting portions of the microfluidic sample channel system 5. The first reagent chamber 11a may contain or be configured to contain at least one capture reagent 33 (capture antibody), and the second reagent chamber 11b may contain or be configured to contain at least one detection reagent 32 (detection antibody). Alternatively, the first reagent chamber 11a may contain or be configured to contain at least one detection reagent 32 (detection antibody), and the second reagent chamber 11b may contain or be configured to contain at least one capture reagent 33 (capture antibody). Alternatively, the first reagent chamber 11a and the second reagent chamber 11b may contain only or be configured to contain one or more (e.g., two, three, four, five, six or more) detection reagents 32 (detection antibodies) or one or more (e.g., two, three, four, five, six or more) capture reagents 32 (capture antibodies). It is even conceivable that one or both reagent chambers may contain both capture and detection reagents 32, 33 simultaneously. In the case of multiplex testing, a reagent chamber may contain or be configured to contain several reagents 32, 33, each binding to a corresponding binding site of multiple analytes 31.

[0134] A fourth section of the microfluidic sample channel system 5 connects the second reagent chamber 11b to the sample release port 4b in the test area 4, where a (treated) sample fluid 30 can be released to provide a flow initiation side 6b for a given test membrane 6 located in a shared recessed test membrane region 4a. The test membrane 6 contacts a waste pad 9 at a flow termination side 6c, positioned in a waste pad region 4d adjacent to the shared recessed test membrane region 4a in the test area 4. The test membrane 6 can be positioned, specifically loosely positioned, clamped, bonded, and / or fixed to the waste pad 9, and can overlap with the waste pad 9. Alternatively, the test membrane 6 contacts the waste pad 9 without overlapping. The sample fluid 30 can then be passively delivered to the waste pad 9 along the overall flow axis 6a of the test membrane 6. In other words, the sample fluid can flow along the overall flow axis 6a.

[0135] The processed sample fluid 30 can be considered as sample fluid 30 that has been processed by at least one of the following steps (but not limited to): aliquoting / dispensing, separating from other elements and / or portions of the sample to which it is applied, labeling the analyte in the sample fluid with a reagent (specifically a detection reagent and / or a capture reagent), diluting the sample fluid, purifying the sample fluid, purifying and / or enriching one or more analytes, dissolving the analyte and / or portions and / or fragments of the sample fluid 30.

[0136] On the other side of the test carrier 2, adjacent to the side of the microfluidic sample channel system 5 located on the test carrier 2, a wash buffer reservoir receiver 14 or a receiving portion / recess (not shown, see Figure 4) for receiving the wash buffer reservoir / blister 13 is provided. The wash buffer reservoir receiver 14 may correspond to a recess, a notch, and / or a hole. The microfluidic wash buffer channel system 12 can deliver and / or guide the wash buffer from the open blister 13 to the wash buffer release port 4c in the test area 4. The blister 13 may be punctured by a needle or sharp element to establish an open state and a fluid connection between the interior of the blister 13 and the microfluidic wash buffer channel system 12. The first part of the microfluidic wash buffer channel system 12 connects the blister 13 to the wash buffer dispensing chamber 15, wherein the first part of the microfluidic wash buffer channel system 12 can be regarded as the first connecting part of the wash buffer dispensing chamber 15, and the wash buffer dispensing chamber 15 can be regarded as a component of the microfluidic wash buffer channel system 12. The wash buffer dispensing chamber 15 has the function of dispensing / dividing the wash buffer into several portions (i.e., dispensing volumes of wash buffer) when the microfluidic test carrier 2 is rotated. The dispensed wash buffer is further delivered through the microfluidic wash buffer channel system 12 to the wash buffer release port 4c in the test area 4. The entire volume of wash buffer in the blister 13 can be divided into several parts (e.g., two, three, four, five, six, or even more parts / dispenses) and provided to the wash buffer release port 4c.

[0137] The microfluidic test carrier 2 shown in Figures 2, 3a, 3b, 4, and 5a has a first side S1 and a second side S2 opposite to the first side S1, respectively. Figures 2, 3a, 3b, and 5a are top views of the corresponding second side S2 of IVD test systems 1a and 1c. Figure 4 is a top perspective view of the first side S1 of IVD test system 1b.

[0138] As shown in Figures 2, 3a, 3b, and 5a, in most cases, the mentioned functional elements of the corresponding microfluidic test carriers 2 of the IVD test systems 1a, 1b, and 1c correspond to recessed elements and / or traces on the corresponding test carrier 2 (specifically, the test carrier core 2c). In these embodiments, the recessed traces are traces in the surface of the second side S2 of the microfluidic test carrier 2, and a second cover plate 2b (not shown, see Figure 4) may be attached and / or fixed to the test carrier core 2c to cover the second side S2 of the corresponding IVD test systems 1a, 1b, and 1c. Therefore, the second cover plate 2b may at least partially seal and / or cover and / or surround the microfluidic sample channel system 5. A first cover plate S1 may cover the opposite side, i.e., the first side of the microfluidic test carrier 2. Therefore, the microfluidic sample channel system 5 may correspond to or include recessed traces in the test carrier core 2c on the second side, and the microfluidic sample channel system 5 may be substantially defined and / or surrounded by the recessed surface of the second side S2 of the test carrier core 2c and the second cover plate 2b. A pathway from the first side S1 via holes, cutouts, and / or recesses (e.g., sample application port 3) in the first cover plate 1a and / or carrier support center member 2c to the microfluidic sample channel system 5 may or may not exist. Alternatively or additionally, a pathway from the second side S2 via holes, cutouts, and / or recesses (e.g., sample application port 3) in the second cover plate 2b to the microfluidic sample channel system 5 may be provided. Alternatively or additionally, the microfluidic sample channel system 5 may correspond to or include recessed traces in one or both of the cover plates 2a and 2b.

[0139] The sample application port 3 may correspond to a hole in one of the cover plates 2a and 2b, that is, the sample application port 3 may correspond to a hole in the first cover plate 2a or the second cover plate 2b. The test carrier core 2c may have the starting point of the microfluidic sample channel system 5, the position of which corresponds to the position of the sample application port 3. In other words, when the corresponding cover plates 2a and 2b are correctly fixed to the test carrier core 2c, the sample application port 3 is positioned exactly above or below the starting point of the microfluidic sample channel system 5.

[0140] Test area 4 may correspond to a recess in the test carrier core 2c. Specifically, test area 4 may correspond to a recess in the test carrier core 2c with a different depth. For example, the recess in test area 4 in the region of sample release port 4b and the region of wash buffer release port 4c may be slightly deeper than the recess in the shared recessed assay membrane region 4a and / or the waste pad region 4d. Further, the recess in the shared recessed assay membrane region 4a may have a different depth than the recess in the waste pad region 4d. For example, the recess in the shared recessed assay membrane region 4a may have a different depth than the recess in the waste pad region 4d because the waste pad 9 and assay membranes 6, 7 may have different thicknesses and / or may overlap in some areas. The size of the recess in test area 4 may be specifically determined to receive the waste pad 9 and one or more assay membranes 6, 7. The depth of the recess can be matched to the thickness of the measuring membranes 6, 7, 8 and / or the waste pad 9, and / or the overlap between the measuring membranes 6, 7, 8 and the waste pad 9.

[0141] Figure 3a is a schematic top perspective view of the IVD assay system 1a according to the example in Figure 2. Figure 3b is a schematic top perspective view of the IVD assay system 1a for multiplex detection, which includes two immunochromatographic assay membranes 6, 7 disposed in the centrifugal microfluidic assay carrier 2 of Figure 2. Figures 3a and 3b are shown adjacent to each other to demonstrate that the same centrifugal microfluidic assay carrier 2 can be used to detect a single analyte using one assay membrane 6 (Figure 3a) and to detect a single analyte using two assay membranes 6, 7 (Figure 3a). Figure 3b ) is used for multiplex detection. In other words, it can be used with a single assay membrane 6 (Fig. 3a) and with two assay membranes 6, 7 ( Figure 3b The centrifugal microfluidic test carrier 2 used for multiplex detection is identical. According to an embodiment, the IVD testing system 1a of Figure 3b is configured for multiplex detection of analytes 31a and 31b on two assay membranes 6 and 7. If the width of the assay membrane is chosen to be smaller than that used with one or two assay membranes, the same centrifugal microfluidic test carrier 2 can even be used for multiplex detection with three, four, five, six, or more assay membranes.

[0142] The first measuring membrane 6 of the IVD testing system 1a for multiplex detection has an overall flow axis 6a, a flow initiation side 6b, and a flow termination side 6c. The first measuring membrane 6 contacts the waste pad 9 at its flow termination side 6c. The second measuring membrane 7 of the IVD testing system 1a for multiplex detection has an overall flow axis 7a, a flow initiation side 7b, and a flow termination side 7c. The second measuring membrane 7 contacts the waste pad 9 at its flow termination side 7c. The first measuring membrane 6 and the second measuring membrane 7 are aligned adjacent to each other, specifically parallel to each other with respect to the overall flow axes 6a and 7a, such that the flow initiation side 6b of the first measuring membrane 6 and the flow initiation side 7b of the second measuring membrane 7 are positioned adjacent to each other, and the flow termination side 6c of the first measuring membrane 6 and the flow termination side 7c of the second measuring membrane 7 are positioned adjacent to each other. The two measuring membranes 6 and 7 share a waste pad 9. The measuring membranes 6 and 7 are separated by a gap 16. Gap 16 can be an air gap and / or a gap without filling membrane material, while the two measuring membranes 6, 7 can be positioned and / or fixed to the substrate. However, it is not required in all embodiments that the two measuring membranes 6, 7 are located on the substrate (specifically a common substrate). The two measuring membranes 6, 7 can, for example, be fixed to one or more waste pads 9 and / or supported by one or more waste pads.

[0143] Figures 8a to 8h show different cuts in the test carrier core 2c of the test carrier according to different embodiments shown in Figures 3a and 3b.

[0144] In Figure 3a, a schematic section line a-a' is shown along the length axis of the measuring membrane 6. Furthermore, a section line A-A' is also indicated that crosses (i.e., is perpendicular to) the section line aa' and the length axis of the measuring membrane 6.

[0145] Figure 8a is a schematic diagram showing a cut along the section line AA' according to an embodiment. The measuring membrane 6 is supported by the bottom / support surface 39 of the recessed measuring membrane region 4a. The recessed measuring membrane region 4a is limited by the recessed wall 38 of the common recessed measuring membrane region 4a indicated on both sides. In other words, the measuring membrane 6 is received and / or embedded in the recessed measuring membrane region 4a. The measuring membrane 6 may be positioned only in and / or clamped in and / or adhered to the bottom / support surface 39 of the recessed measuring membrane region 4a.

[0146] Figures 8b, 8c, and 8d are schematic diagrams illustrating cuts along section line aa' according to different embodiments. Figure 8b shows the measuring membrane 6 and waste pad 9 positioned in the test area 4 and flush with each other. The bottom / support surface 39 of the test area 4 (specifically the recessed measuring membrane area 4a) is a flat surface / flat plane without steps. In another embodiment, the measuring membrane 6 and waste pad 9 may overlap each other and / or may be fluidly connected to each other via another pad (not shown). Compared to Figure 8b, Figures 8c and 8d show steps in the bottom / support surface 39 of the test area 4, wherein in Figure 8c, the horizontal level of the bottom / support surface 39 supporting the waste pad 9 is lower than the horizontal level of the bottom / support surface 39 supporting the measuring membrane 6, and wherein in Figure 8d, the horizontal level of the bottom / support surface 39 supporting the measuring membrane 6 is lower than the horizontal level of the bottom / support surface 39 supporting the waste pad 9. In both cases, a step is provided between the area primarily supporting the measuring membrane 6 and the area primarily supporting the waste pad 9. Preferably, the waste pad 9 and the measuring membrane 6 overlap each other to provide a reliable fluid connection between them. The depth of the step can be from 0.1 mm to 3 mm, specifically from 0.5 mm to 2 mm.

[0147] Figures 8e and 8f are schematic diagrams showing cuts along the section line BB' according to different embodiments. Figure 8eFigure 8f is a schematic diagram showing a cut along the section line BB' according to one embodiment. Measuring membranes 6 and 7 are in contact with each other, with a distance of 0 mm between them. Measuring membranes 6 and 7 are supported by the bottom / support surface 39 of the recessed measuring membrane region 4a. The recessed measuring membrane region 4a is limited by the recessed wall 38 of the shared recessed measuring membrane region 4a indicated on both sides. In other words, measuring membranes 6 and 7 are received and / or embedded in the recessed measuring membrane region 4a. Measuring membranes 6 and 7 may be positioned only in and / or clamped in and / or adhered to the bottom / support surface 39 of the recessed measuring membrane region 4a. Figure 8f is a schematic diagram showing a cut along the section line BB' according to one embodiment. Compared to Figure 8e, in this embodiment, measuring membranes 6 and 7 are not in direct contact with each other and / or have a gap between them with a distance d > 0 mm. The bottom / support surface 39 of the recessed measuring membrane region 4a is a flat, stepless surface, ensuring no obstruction between the first measuring membrane 6 and the second measuring membrane 7. Generally, in all illustrated embodiments, the bottom / support surface 39 of the recessed measuring membrane region 4a can be a flat, stepless surface, ensuring no obstruction between the two measuring membranes 6, 7.

[0148] Figures 8g and 8h are schematic diagrams showing cuts along the section line bb' according to different embodiments. Figure 8g The embodiment of Figure 8h corresponds to the embodiment of Figure 8d, except that it has two measuring membranes 6, 7 instead of just a single measuring membrane 6. The embodiment of Figure 8h corresponds to the embodiment of Figure 8d, except that it has two measuring membranes 6, 7 instead of just a single measuring membrane 6. Preferably, the shared recessed measuring membrane region 4a (specifically shown in Figures 8e to 8h) is a planar surface without walls and / or protrusions between the two measuring membranes 6, 7. However, if protrusions are to be provided between the two measuring membranes 6, 7, or generally between at least two or more measuring membranes (e.g., for better alignment and / or to support alignment of the measuring membranes 6, 7 in the test area), the protrusions may not have the same height as the measuring membranes (specifically, a lower height) and / or may not have the same length as the measuring membranes 6, 7 (specifically, shorter than the measuring membranes 6, 7). Specifically, the recessed measuring membrane region 4a should not correspond to a substantially bifurcated path of fluid before reaching measuring membranes 6 and 7, but should correspond to a common / shared reservoir from which fluid can be received by measuring membranes 6 and 7 (specifically substantially simultaneously).

[0149] The width w of the (shared) recessed measuring membrane region 4a can be between approximately 0.1 cm and 3 cm, specifically between approximately 0.5 cm and 2 cm, and more specifically between approximately 0.8 cm and 1.5 cm. The edge-to-edge distance d between two adjacent measuring membranes can be between approximately 0 and 1 cm, specifically between approximately 0.1 and 5 mm, and more specifically between approximately 0.3 and 1 mm. The width w of the (shared) recessed measuring membrane region 4a can match the width of one measuring membrane 6, or—if multiple measuring membranes are provided—the combined width of the measuring membranes 6 and 7. Specifically, the width w of the (shared) recessed measuring membrane region 4a may correspond to approximately 1 to 2 times the width of a measuring membrane 6, specifically approximately 1.1 to 1.7 times, and more specifically approximately 1.2 to 1.5 times, or—if multiple measuring membranes are provided—a multiple of the combined width of the provided measuring membranes 6, 7.

[0150] The length l of the (shared) recessed measuring membrane region 4a, including the area for receiving the waste pad 9, can be between approximately 1 cm and 7 cm, specifically between approximately 2 cm and 5 cm, and more specifically between approximately 2.5 cm and 3 cm. The length l of the (shared) recessed measuring membrane region 4a, including the area for receiving the waste pad 9, can match the length of the measuring membrane 6 and the length of the waste pad 9. Specifically, the length l of the (shared) recessed measuring membrane region 4a, including the area for receiving the waste pad 9, can correspond to approximately 1 to 1.5 times the length of one of the measuring membranes 6 and 7 arranged in combination with the waste pad 9, specifically between approximately 1 to 1.3 times, and more specifically between approximately 1 to 1.2 times. The length l1 of the (shared) recessed measuring membrane region 4a (excluding the length l2 of the region receiving waste pad 9) can be between approximately 0.5 cm and 4 cm, specifically between approximately 1 cm and 3 cm, and more specifically between approximately 1.5 cm and 2 cm. The length l2 of the region receiving waste pad 9 can be between approximately 0.2 cm and 3 cm, specifically between approximately 0.5 cm and 2 cm, and more specifically between approximately 0.8 cm and 1.2 cm. Figure 4This is an exploded top perspective view of an IVD testing system 1b according to another embodiment, which includes three immunochromatographic assay membranes located in a centrifuged microfluidic test carrier 2 for multiplex detection of analytes. The test carrier core 2c of FIG4 is shown from the side opposite to FIG2, 3a, and 3b (top views viewed from the second side S2) (top views viewed from the first side S2). The test carrier 2 may have all the features and advantages described in conjunction with FIG2, 3a, and 3b, and vice versa. The IVD testing system 1b of FIG4 includes a test carrier 2 having a test carrier core 2c, which can be produced by injection molding, having a first cover plate 2a for at least partially covering the first side S1 of the test carrier core 2c, and a second cover plate 2b for at least partially covering the second side S2 of the test carrier core 2c. The test carrier core 2c is sandwiched between the first cover plate 2a and the second cover plate 2b. This configuration, with a first cover plate 2a and a second cover plate 2b, can also be applied. Figure 2 The embodiments shown in Figures 3a and 3b. Alternatively, it is conceivable that the test carrier core 2c is covered only on the first side S1 by the first cover plate 2a and the second cover plate 2b is not required. The first cover plate 2a can serve as a top cover on which a label is placed, the label having information and / or specifications regarding test type, sample type, sample quantity, brand, charge number, model, design elements, barcode, patient and / or sample information and / or identification notes, etc. The second cover plate 2b can serve as a sealing cap. The first cover plate 2a includes cutouts and / or holes to provide access to the blister 13, the microfluidic sample channel system 5, and / or the assay membranes 6, 7, 8.

[0151] The IVD testing system 1b of Figure 4 further includes a first assay membrane 6 configured to detect at least one first analyte; a second assay membrane 7 configured to detect at least one second analyte; and a third assay membrane 8 configured to detect at least one third analyte. Each assay membrane 6, 7, 8 can be configured to detect an independent analyte. In other words, the first, second, and third analytes can be different analytes. Alternatively, at least one of the assay membranes can be configured to detect the same analyte as another assay membrane, but within a different sensitivity range.

[0152] The IVD testing system 1b of Figure 4 further includes a shared waste pad 9 and a blister / wash buffer reservoir 13 for storing wash buffer. The IVD testing system 1b shown in Figure 4 can be considered a cartridge that can be used in the IVD testing apparatus 20, for example, as shown in Figure 6a.

[0153] The dimension of the longest side of the test carrier 2 can be determined to be between approximately 3 cm and 12 cm, specifically between approximately 4 cm and 10 cm, and more preferably between approximately 5 cm and 7 cm. The dimension of the shorter side of the test carrier 2 (measured perpendicular to the longest side and possibly parallel to the overall flow axis of the measuring membranes 6, 7, 8 when the test carrier is inserted into / on the cartridge) can be determined to be between approximately 2.5 cm and 10 cm, specifically between approximately 3 cm and 7 cm, and more preferably between approximately 4 cm and 6 cm. The thickness of the test carrier 2 (with two cover plates 2a, 2b) can be between approximately 0.1 cm and 1 cm, preferably between approximately 0.3 cm and 0.5 cm.

[0154] If only two measuring membranes 6 and 7 are provided in the test assembly 1a, as shown in Figure 3b, the widths 6d and 7d of the first measuring membrane 6 and the second measuring membrane 7, measured in a direction perpendicular to the respective overall flow axes 6a and 7a, can be approximately 1.3 mm to 2.3 mm, specifically approximately 1.5 mm to 2 mm.

[0155] If three measuring membranes 6, 7, and 8 are set in the test assembly 1b, as shown in Figure 4, the widths 6d, 7d, and 8d of the first measuring membrane 6, the second measuring membrane 7, and the third measuring membrane 8, measured in directions perpendicular to their respective overall flow axes 6a, 7a, and 8a, can be approximately 0.8 mm to 1.5 mm, specifically approximately 1 mm to 1.3 mm.

[0156] Figure 5a is a schematic top perspective view of the IVD testing system 1a according to the embodiment of Figure 3b. Figure 5b is a schematic diagram of the first capture mechanism of the first assay membrane 6, which is provided as the immunochromatographic assay element of Figure 5a. Figure 5c is a schematic diagram of the second capture mechanism of the second assay membrane 7, which is provided as the immunochromatographic assay element of Figure 5a. Together with Figures 5a to 5c, multiplex assays are described in more detail below.

[0157] In this embodiment, the first analyte 31a, detectable on the first assay membrane 6, may correspond to cardiac troponin T (TnT). In this embodiment, the second analyte 31b, detectable on the second assay membrane 7, may correspond to NTproBNP. The first reagent chamber 11a contains a first capture reagent 33a and a first capture mechanism 34a disposed on the first assay membrane 6, the first capture reagent binding to the binding site of TNT; and a second capture reagent 33b and a second capture mechanism 34b disposed on the second assay membrane 7, the second capture reagent binding to the binding site of NTproBNP. In this embodiment, the first capture reagent 33a may correspond to a dried capture Ab for the first analyte 31a, such as a biotinylated Ab, such as MAB-M-11.7-F(ab')2-Bi, and the first capture mechanism 34a may correspond to streptavidin. In this embodiment, the second capture reagent 33b may correspond to a dried capture Ab for the second analyte 31b, such as a digoxigenin-ligated Ab, like MAB-S-1.21.3-F(ab')2-Dig, and the second capture mechanism 34b may correspond to anti-dig. The second reagent chamber 11b contains a first detection reagent 32a and a second detection reagent 32b, the first detection reagent binding to another binding site of TNT and the second detection reagent binding to another binding site of NTproBNP. In this embodiment, the first detection reagent 32a may correspond to a dried capture Ab for the first analyte 31a, such as a fluorescent Ab, like MAB-M-17.57.511-IgG-JG9-Lx. In this embodiment, the second detection reagent 32b may correspond to a dried detection Ab for the second analyte 31b, such as a fluorescent Ab, like MAB-M-18.4.34-IgG-JG9-Lx.

[0158] Figure 5b shows a first assay membrane 6 for detecting the first analyte 31a, and Figure 5c shows a second assay membrane 7 for detecting the second analyte 31b. The two assay membranes 6 and 7 are similar to the examples shown in Figures 1a and 1b illustrating a known point-of-care (POC) test concept. In this example, a whole blood sample may represent the sample fluid 30 received from the patient. In this example, the sample fluid 30 contains both the first analyte 31a and the second analyte 31b to demonstrate the principle. Of course, the sample fluid 30 can also be tested if it does not contain one or more target analytes 31. In this case, the test result for the target-specific analyte 31 is negative.

[0159] Plasma 30a is extracted from whole blood sample 30 in plasma separation element 11 by centrifugation, and / or plasma is extracted from whole blood sample by plasma separation element. In this example, plasma 30a contains first analyte 31a and second analyte 31b. The centrifuge is either started to rotate at a frequency suitable for moving plasma 30a from plasma separation element 11 to capture reagent chamber 11a, or the delivery process relies solely on the capillary force of microfluidic sample channel system 5. Plasma 30a is mixed with dried first capture reagent 33a and dried second capture reagent 33b and incubated for a predetermined amount of time during which the centrifuge does not rotate, slows down, or stops. Plasma 30a dissolves capture reagents 33a and 33b, which then couple to the binding sites of first analyte 31a and second analyte 31b, respectively, thus labeling analytes 31a and 31b for subsequent binding processes. The centrifuge can be restarted to rotate at a frequency suitable for moving plasma 30a from the capture reagent chamber 11a to the detection reagent chamber 11b, or the delivery process can rely solely on the capillary force of the microfluidic sample channel system 5. Plasma 30a dissolves detection reagents 32a and 32b, which then couple to other binding sites of the first analyte 31a and the second analyte 31b, thus labeling analytes 31a and 31b for subsequent detection.

[0160] Measuring membranes 6 and 7 each have an overall flow axis 6a, 7a, a flow initiation side 6b, 7b, and a flow termination side 6c, 7c. In some cases, measuring membranes 6 and 7 may be in contact with one or more waste pads 9 at the flow termination side 6c, 7c. Figures 5b and 5c show measuring membranes 6 and 7 after the application of serum 30a and after the labeled analytes 31a, 31b, 31c have bound to and / or been bound by the measuring membranes, respectively.

[0161] Plasma 30a, containing labeled analytes 31a, 31b, and 31c, is supplied via sample release port 4b to assay membranes 6 and 7 located at corresponding flow initiation sides 6b and 7b. At least one portion (specifically a fraction and / or component) of plasma 30a is passively transported substantially along the overall flow axis 6a and 7a of each assay membrane 6 and 7 from the flow initiation side 6b and 7b to the flow termination side 6c and 7c by capillary force. Each assay membrane 6 and 7 includes three lines 35, 36, and 37: a control line 35, a calibration line 36, and a capture and test result line 37. The control line 35 (assay control) of the first membrane 6 is pre-coated with the target first analyte 31a, and the control line 35 of the second membrane 7 is pre-coated with the target second analyte 31b. The calibration line 36 (instrument calibration) of the first membrane 6 is pre-coated with a fluorescent label including the first detection reagent 32a (detection antibody), and the calibration line 36 (instrument calibration) of the second membrane 7 is pre-coated with a fluorescent label including the second detection reagent 32b. The capture and test result line 37 of the first membrane 6 is coated with the first capture mechanism 34a (streptavidin). The capture and test result line 37 of the second membrane 7 is coated with the second capture mechanism 34b (anti-dig).

[0162] Serum 30a is applied to the corresponding flow initiation sides 6b and 7b of the assay membranes 6 and 7, and thus first passes through the capture and test result line 37. At this line, if labeled analytes 31a and 31b are present in serum 30a, the analytes 31a and 31b sandwiched between the two reagents 31a, 31b, 32a, and 32b are specifically bound / captured by capture antibodies 33a and 33b via binding to the first and second capture mechanisms 34a and 34b. The residual portion of serum 30a passes through the calibration line 36, where no further binding occurs; then, the residual portion passes through the control line 35, where residual unbound detection reagents 32a and 32b, including detection antibodies, are at least partially and specifically bound via binding sites of analytes 31a and 31b provided on the pre-coated control line 35.

[0163] Control line 35 controls whether serum 31 is mixed with reagents 32a and 32b by capturing unbound test reagents 32a and 32b. Control line 35 is particularly important if the test result is negative, as the negative result may be due to the absence of analytes 31a and 31b, but it could also be due to the absence of fluorescent labels 32a and 32b (especially false negatives). Calibration line 36 is used for calibration purposes and provides a pre-coated area with fluorescent labels for use with detection antibodies 32a and 32b to label analytes 31a and 31b for detection.

[0164] Finally, the assay membranes 6 and 7 are washed with buffer supplied via the wash buffer release port 4c to remove unbound reagents 31a, 31b, 32a, and 32b, and CCD images can be recorded to read the test results. The test results of assay membranes 6 and 7, schematically shown in Figures 5b and 5c, will be positive for TnT and NTproBNP.

[0165] Figure 6a is a schematic diagram of an IVD testing apparatus 20 including IVD testing systems 1a and 1b according to one embodiment. The IVD testing apparatus 20 includes a centrifuge 21 with a cartridge support plate 23 configured to rotate about a centrifugal axis 25 via a motor of the centrifuge 21. The IVD testing apparatus 20 further includes a controller 22 configured to control the rotation of the motor of the centrifuge 21 and the cartridge support plate 23. The IVD testing apparatus 20 includes at least one cartridge 24, which includes the IVD testing system 1a and / or corresponds to the IVD testing system 1a according to the embodiments described herein, wherein the cartridge 24 is configured to be fixed to the cartridge support plate 23 so as to rotate together with the cartridge support plate 23 rotating about the centrifugal axis 25. Preferably, a plurality of cartridges 24 (such as two, three, four, five, or even more cartridges 24) may be fixed to the cartridge support plate 23. If each cartridge 24 provides a different multiplex test for detecting two different analytes, then providing three cartridges 24 allows for the detection of six analytes. Alternatively, the multiplex test can be the same, and the cartridges 24 can be loaded with samples from different patients.

[0166] Figure 6b is a top view of the cartridge support plate 23 of the three cartridges 24 of the IVD testing apparatus 20 of Figure 6a according to one embodiment. As can be seen from the above, the cartridges 24 are evenly distributed around the centrifugal axis 25.

[0167] Figure 7 is a schematic diagram of a method 100 for performing multiple diagnostic assays according to one embodiment. Therefore, the microfluidic cartridge allows control over each step of a single assay, resulting in superior and laboratory-like assay performance, such as high sensitivity, high precision, and high accuracy.

[0168] The method 100 for performing multiple diagnostic assays can be performed using an IVD testing system 1a to perform multiple diagnostic assays as described in one of the embodiments. The method 100 may include the step of providing an IVD testing apparatus 20 as described with FIG. 7. The IVD testing apparatus 20 may include: a centrifuge 21 having a cartridge support plate configured to rotate about a centrifugal axis 25 via a motor of the centrifuge 21; a controller 22 configured to control the rotation of the motor of the centrifuge 21 and the cartridge support plate 23; and at least one cartridge 24 comprising the IVD testing system 1a, and / or corresponding to the IVD testing system 1a according to the embodiments described herein, wherein the cartridge 24 is configured to be fixed to the cartridge support plate 23 so as to rotate together with the cartridge support plate 23 rotating about the centrifugal axis 25.

[0169] Method 100 may further include: positioning 102 and / or securing at least one cartridge 24 to a cartridge support plate 23; rotating 103 the cartridge support plate 23 with the cartridge 24 about a rotation axis 25 in a centrifuge 21 at at least one first predetermined rotation frequency, the first predetermined rotation frequency being configured to guide at least a portion 30, 31 of sample fluid 30 at least partially from the sample application port 3 through the microfluidic sample channel system 5 to the sample release port 4b to provide 105 at least a portion 30, 31 of sample fluid 30 to a first assay membrane 6 and a second assay membrane 7, enabling multiple diagnostic assays 104; and detecting 105 whether the first assay membrane indicates the presence of a first analyte and / or whether the second assay membrane indicates the presence of a second analyte. The rotation procedure (sequence and frequency) may be optimized for the cartridge design so that the component design based on the microfluidic sample channel system 5 precisely performs each function.

[0170] In some embodiments, the method may include at least one of the following steps: Specifically, the step “rotating 103 of a cartridge support plate 23 with cartridge 24 about a rotation axis 25 in a centrifuge 21 at at least a first predetermined rotation frequency, the first predetermined rotation frequency being configured to guide at least a portion 30, 31 of sample fluid 30 at least partially from the sample application port 3 through the microfluidic sample channel system 5 to the sample release port 4b to provide 105 at least a portion 30, 31 of sample fluid 30 to the first assay membrane 6 and the second assay membrane 7, thereby enabling multiple diagnostic assays 104” may include or involve further steps. For example, rotation 103 may correspond to the performance of a rotation procedure, wherein rotating the cartridge support plate 23 with cartridge 24 about a rotation axis 25 in a centrifuge 21 at at least a first predetermined rotation frequency allows at least a portion of the sample fluid in at least a segment to be moved from the sample application port to the plasma separation element 10. The performance of the rotation process may include or involve rotation at at least one first predetermined rotation frequency or another predetermined rotation frequency, the rotation being configured to perform plasma separation. The performance of the rotation process may include or involve rotation at at least one first predetermined rotation frequency or another predetermined rotation frequency, the rotation being configured to move at least a portion of the plasma from the plasma separation element 10 to the first reagent chamber 11a. This optional step may accelerate capillary-driven delivery. The performance of the rotation process may include or involve stopping rotation, and allowing binding reactions between the first analyte 31a and the first capture reagent 33a, and between the second analyte 31b and the second capture reagent 33b, if the first analyte 31a and the second analyte 31b are present in the sample fluid 30.

[0171] The performance of the rotation procedure may include or involve rotation at at least one first predetermined rotation frequency or another predetermined rotation frequency, the rotation being configured to move at least a portion of the plasma from the first reagent chamber 11a to the second reagent chamber 11b. This optional step may accelerate capillary-driven delivery. The performance of the rotation procedure may include or involve stopping the rotation, and allowing binding reactions between the first analyte 31a and the first detection reagent 32a, and between the second analyte 31b and the second detection reagent 32b, if the first analyte 31a and the second analyte 31b are present in the sample fluid 30.

[0172] The performance of the rotation process may include or involve rotating a cartridge support plate 23 with cartridge 24 in a centrifuge 21 at at least one first predetermined rotation frequency or another predetermined rotation frequency, the first predetermined rotation frequency or the other predetermined rotation frequency being configured to provide at least a portion 30, 31 of sample fluid 30 to a first measuring membrane 6 and a second measuring membrane 7, such that a first portion of at least a portion 30, 31 of sample fluid 30 flows along the overall flow axis 6a of the first measuring membrane 6, and thus, if a first analyte 31a is present in the sample fluid 30, a capture process of the first analyte 31a on the first measuring membrane 6 is permitted, and a second portion of at least a portion 30, 31 of sample fluid 30 flows along the overall flow axis 7a of the second measuring membrane 7, and thus, if a second analyte 31b is present in the sample fluid 30, a capture process of the second analyte 31b on the second measuring membrane 7 is permitted.

[0173] The performance of the rotation procedure may include or involve opening the wash buffer reservoir 13. The performance of the rotation procedure may include or involve rotating the cartridge support plate 23 with cartridge 24 in the centrifuge 21 at at least one first predetermined rotation frequency or another predetermined rotation frequency, the rotation being configured to guide at least one portion of the wash buffer from the wash buffer reservoir 13 via the microfluidic wash buffer channel system 12 to the wash buffer release port 4c to provide at least one portion of the wash buffer to the first assay membrane 6 and the second assay membrane 7. A final step may include or involve a fractionation step, wherein the rotation frequency is configured to divide the wash buffer into several portions by means of a wash buffer fractionation chamber. Alternatively, the wash buffer may be delivered / guided by capillary force through at least a portion of the microfluidic wash buffer channel system 12 without rotation.

[0174] The principles, effects, advantages and embodiments of the present invention are generally described below.

[0175] IVD testing systems 1a and 1b are shown to include a waste pad 9, but in other embodiments, they may not provide a waste pad 9. However, IVD testing systems 1a and 1b are shown to include two reagent chambers 11a and 11b, at least one of which may be divided into multiple sub-chambers for containing a first reagent in one sub-chamber and a second reagent in another sub-chamber. In other embodiments, IVD testing systems 1a and 1b may also include only one reagent chamber 11. IVD testing systems 1a and 1b may also include more than two reagent chambers 11a and 11b, for example, three, four, five, six, or more reagent chambers. IVD testing systems 1a and 1b are shown as including recessed and open microfluidic channels 5 and channel elements 10, 11, 12, and 15, provided no cover plate is applied; however, in other embodiments, the microfluidic channels 5 and channel elements 10, 11, 12, and 15 may also correspond to substantially and / or at least partially closed channel structures, which do not require a cover plate to control the flow of microfluidics through the channels. IVD testing systems 1a and 1b are shown as including a recessed test area 4; however, in other embodiments, the test area 4 may not be recessed.

[0176] IVD testing systems 1a and 1b are shown to include a wash buffer reservoir receiver 14 corresponding to a cutout in test carrier 2c. In other embodiments, the wash buffer reservoir receiver 14 may correspond to a recess and / or region to provide space for a wash buffer reservoir 13. In other embodiments, the wash buffer reservoir 13 may be integrated into test carriers 2 and 2c. Test carriers 2 and 2c may, for example, include a chamber for containing wash buffer. In other embodiments, the wash buffer reservoir 13 may be fixedly attached to test carriers 2 and 2c. The wash buffer reservoir 13 may be bonded, heat-fused, and / or welded to test carriers 2 and 2c. IVD testing systems 1a and 1b are shown to include two or three assay membranes 6, 7, and 8 positioned parallel to each other relative to the respective overall flow axes 6a, 7a, and 8a. In other embodiments, more than three assay membranes may be provided. In other embodiments, the assay membranes may be positioned adjacent to each other but not parallel aligned. For example, measuring membranes 6, 7, and 8 can be positioned in a star-shaped and / or fan-shaped assembly / or orientation relative to each other, characterized by an angle not 0°, 180°, and / or 360° enclosed by the length axes of the two measuring membranes and / or the overall flow axis.

[0177] A microfluidic assay carrier can be described as a microfluidic cartridge design that allows for the parallel execution of multiple determinations of two, three, or even more analytes using only a single toolbox-like approach. Sample volumes can range from approximately 10 µL to 100 µL, specifically from approximately 15 µL to 70 µL, and more specifically from approximately 30 µL to 35 µL, of sample fluids such as whole blood. Depending on the number of analytes to be multiplexed, two, three, or more assay membranes are placed in the microfluidic assay carrier 2. For example, in a single analyte assay, the membrane width of one assay membrane (6d, 7d, 8d) can be approximately 3 mm to 4 mm; in a two-analyte assay, the membrane width of one of the two assay membranes can be approximately 1.5 mm to 2 mm; and in a three-analyte assay, the membrane width of one of the three assay membranes can be approximately 1 mm to 1.3 mm. All membranes may be of the same length, for example, approximately 17 mm. When sandwich immunoassay is assumed to be used as the test format, the capture region of the first analyte may contain streptavidin for capturing biotinylated antibodies, the capture region of the second analyte may contain anti-dig for capturing dig-labeled antibodies, and the capture region of the third analyte may contain analyte-specific antibodies for directly capturing the corresponding analyte.

[0178] The following text provides some general definitions.

[0179] Serum is the clear, liquid portion of blood that can be separated from clotted blood. Plasma corresponds to the clear, liquid portion of blood containing blood cells. Serum differs from plasma, which is the liquid portion of normal, unclotted blood containing red blood cells, white blood cells, and platelets. Clotting causes the difference between serum and plasma. As used herein, the term "whole blood" includes all components of blood, such as white blood cells and red blood cells, platelets, and plasma.

[0180] "Detection analyte" can refer to the qualitative detection of the presence of at least one target analyte. "Detection analyte" can specifically include the meaning of quantitative determination of the amount of analyte, as primarily used herein. As used herein, the term "determining" the level of a target analyte refers to the quantitative or qualitative determination of the target analyte, such as determining or measuring the amount of the target analyte in a sample.

[0181] In this context, "the presence of at least one target analyte" encompasses the qualitative assessment of the target analyte by means of its absolute value, and / or its relative signal value relative to internal standards and / or references and / or other analytes, and / or its concentration limit matching its concentration.

[0182] In this context, “quantity” or “quantity value” encompasses absolute quantity, relative quantity, or concentration, as well as any value or parameter associated with or derived from it.

[0183] In the context of this disclosure, the terms "analyte," "analyte molecule," or "one or more target analytes" are used interchangeably to refer to a chemical substance to be analyzed by mass spectrometry. Chemical substances suitable for analysis by mass spectrometry, i.e., analytes, can be any kind of molecule present in a living organism, including but not limited to nucleic acids (e.g., DNA, mRNA, miRNA, rRNA, etc.), amino acids, peptides, proteins (e.g., cell surface receptors, cytoplasmic proteins, etc.), metabolites or hormones (e.g., testosterone, estrogen, estradiol, etc.), fatty acids, lipids, carbohydrates, steroids, ketone steroids, open-ring steroids (e.g., vitamin D), molecules characterized by a modification of another molecule (e.g., a sugar moiety or phosphoryl residue on a protein, a methyl residue on genomic DNA), or substances already in vivo (e.g., therapeutic drugs, abused drugs, toxins, etc.) or metabolites of such substances. Such analytes can be used as biomarkers. In the context of this invention, the term "biomarker" refers to a substance within a biological system that serves as an indicator of the biological state of said system.

[0184] List of reference numerals

[0185]

Claims

1. An in vitro diagnostic (IVD) testing system (1a, 1b) for performing multiple diagnostic assays, wherein the IVD testing system (1a, 1b) comprises: Test carrier (2), which includes: The sample application port (3) is configured to receive sample fluid (30); The test area (4) includes a shared recessed measurement membrane region (4a) and a sample release port (4b) for releasing at least one portion (30, 30a) of the sample fluid (30) into the shared recessed measurement membrane region (4a); and A microfluidic sample channel system (5) is configured to guide at least one portion (30, 30a) of the sample fluid (30) from the sample application port (3) to the sample release port (4b); The IVD testing system (1a, 1b) further includes: A first assay membrane (6), positioned in the common recessed assay membrane region (4a), is configured to receive a first portion (30, 30a) of the sample fluid (30) from the sample release port (4b) and indicate at least one first analyte (31a) in the sample fluid (30); and A second assay membrane (7), located adjacent to the first assay membrane (6) in the shared recessed assay membrane region (4a), is configured to receive a second portion of the at least one (30, 30a) of the sample fluid (30) from the sample release port (4b), and to indicate at least one second analyte (31b) in the sample fluid (30) and / or to indicate at least one first analyte (31a) in the sample fluid (30) with a sensitivity range different from that of the first assay membrane (6).

2. The IVD testing system (1b) according to claim 1, further comprising: A third assay membrane (8), located adjacent to the first assay membrane (6) and / or the second assay membrane (7) in the recessed assay membrane region (4a), is configured to receive a third portion of the at least one (30, 30a) of the sample fluid (30) from the sample release port (4b) and indicate a third analyte in the sample fluid (30).

3. The IVD testing system (1a, 1b) according to claim 1 or 2, wherein... The first assay membrane (6) captures the first analyte (31a) labeled with the first capture reagent (33a) by means of the first capture mechanism (34a); The second assay membrane (7) captures the second analyte (31a) labeled with a second capture reagent (33b) different from the first capture mechanism (34a) by means of a second capture mechanism (34b), optionally wherein The first capture reagent (33a) and / or the second capture reagent (33b) comprises one of a biotinylated antibody or a digoxigenin-labeled antibody, and optionally wherein... The first capture mechanism (34a) and / or the second capture mechanism (34b) comprises one of the following: streptavidin for capturing the first analyte (31a) or the second analyte (31b) via the biotinylated antibody; anti-dig for capturing the first analyte or the second analyte via the dig-tagged antibody; a DNA-based system; and / or an analyte-specific antibody for directly capturing the first analyte (31a) or the second analyte (31b).

4. The IVD testing system (1a, 1b) according to any one of the preceding claims, wherein the first measuring membrane (6) and the second measuring membrane (7) each have an overall flow axis (6a, 7a), the overall flow axis being defined by a flow initiation side (6b, 7b) and a flow termination side (6c, 7c) opposite to the flow initiation side (6b, 7b), the corresponding overall flow axis (6a, 7a) passing through the flow initiation side and the flow termination side, and wherein the overall flow axis (6a) of the first measuring membrane (6) and the overall flow axis (7a) of the second measuring membrane (7) are aligned in parallel, such that the corresponding flow initiation side (6b, 7b) is positioned as the sample release port (4b) closest to the test area (4).

5. The IVD testing system (1a, 1b) according to claim 4, further comprising at least one waste pad (9) positioned to contact the first test membrane (6) and / or the second test membrane (7) at a corresponding flow end side (6c, 7c).

6. The IVD testing system (1a, 1b) according to any one of the preceding claims, wherein the test carrier includes a plasma separation element (10), the plasma separation element being integrated with the microfluidic sample channel system (5) and configured to extract plasma (30a) from the sample fluid (30) when the sample fluid includes whole blood, optionally wherein the microfluidic sample channel system (5) is configured to guide at least one portion of the sample fluid (30) from the sample application port (3) to the plasma separation element (10) and guide at least one portion of the plasma (30a) from the plasma separation element (10) to the release port (4b).

7. The IVD testing system (1a, 1b) according to any one of claims 3 to 6, wherein the test carrier (2) further comprises At least one reagent chamber (11), integrated with a microfluidic sample channel system (5), is provided for containing at least one reagent (32, 33) configured to bind to at least one of the analytes (31, 31a, 31b), specifically wherein the at least one reagent chamber (11) includes At least one capture reagent chamber (11a) that contains the first capture reagent (33a) and / or the second capture reagent (33b).

8. The IVD testing system (1a, 1b) according to claim 7, wherein the at least one reagent chamber (11) comprises At least one detection reagent chamber (11b) for containing at least one first detection reagent (32a) configured to bind with the first analyte (31a) and / or for containing a second capture reagent (32b) configured to bind with the second analyte (31b).

9. The IVD testing system (1a, 1b) according to any one of the preceding claims. If only two measuring membranes (6, 7) are provided in the test assembly, then the first measuring membrane (6) and the second measuring membrane (7) each have a width of approximately 1.5 mm to 2 mm, 6d and 7d, measured in a direction perpendicular to the respective overall flow axis (6a, 7a); or If three measuring membranes (6, 7, 8) are provided in the test assembly, the first measuring membrane (6), the second measuring membrane (7) and the third measuring membrane (8) each have a width of approximately 1 mm to 1.3 mm, 6d, 7d, 8d, measured in a direction perpendicular to the respective overall flow axis (6a, 7a).

10. The IVD testing system (1a, 1b) according to any one of the preceding claims, wherein the test area (4) includes a wash buffer release port (4c), and wherein the test carrier (2) further includes A microfluidic wash buffer channel system (12) is used to guide the wash buffer; A wash buffer reservoir receiving section (14) for accommodating and / or receiving a wash buffer reservoir (13); the test carrier optionally further includes The washing buffer reservoir (13) is configured to be fluidly connected to the washing buffer release port (4c) via the microfluidic washing buffer channel system (12) for providing a predetermined amount of washing buffer to the assay membrane at the corresponding flow initiation side.

11. The IVD testing system (1a, 1b) according to any one of the preceding claims, wherein the first analyte (31a) and / or the second analyte (31b) comprises at least one of cardiac analytes, specifically cTropT, NTproBNP, D-dimer, PCT, BM2, BM3, creatinine, potassium, and sodium chloride.

12. The IVD testing system (1a, 1b) according to any one of claims 8 to 11, wherein the multiple diagnostic assay corresponds to an immunochromatographic assay, and the at least one first detection reagent (32a) and / or at least one second detection reagent (32b) comprises a fluorescent molecule.

13. The IVD testing system (1a, 1b) according to any one of the preceding claims, wherein the test carrier (2) corresponds to a centrifugal microfluidic test carrier, and wherein the microfluidic channel system (5) is configured to guide at least one portion of the sample fluid (30) from the sample application port (3) to the sample release port (4b) under the influence of centrifugal force.

14. An IVD testing device (20), comprising: Centrifuge (21) having a cartridge support plate configured to rotate about a centrifugal axis (25) by a motor of the centrifuge (21); A controller (22) configured to control the rotation of the motor of the centrifuge (21) and the cartridge support plate (23); At least one cartridge (24) comprising and / or corresponding to the IVD testing system (1a, 1b) according to claim 13, wherein the cartridge (24) is configured to be fixed to the cartridge support plate (23) to rotate together with the rotating cartridge support plate (23) about the centrifugal axis (25).

15. A method (100) for performing multiple diagnostic assays, the method comprising the following steps: Provide (101) the IVD testing equipment (20) according to claim 14; Position (102) and / or fix the at least one cartridge (24) on the cartridge support plate (23); The sample fluid (30) is initially delivered to the first measuring membrane (6) and the second measuring membrane (7) through the microfluidic sample channel system (103, 104); Detection (105) whether the first assay membrane indicates the presence of the first analyte and / or whether the second assay membrane indicates the presence of the second analyte.

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