MXPA06000346A - Device and method for simultaneously identifying blood group antigens - Google Patents

Abstract

The invention relates to a device for simultaneously, qualitatively or quantitatively identifying a number of analytes in a liquid sample, comprising a membrane (2) with:a charging zone (5) for applying the liquid sample;at least two indicator zones, which can interact with the analyte(s), and at least one absorption area (3), which absorbs the liquid after passing the indicator zones, whereby the indicator zones are located between the charging zone (5) and an absorption area (3). The invention is characterized in that the flowing directions from the charging zone (5) through the respective indicator zones to an absorption area (3) (flow paths) are essentially parallel, and at least two different flow paths exist. The invention also relates to a method for identifying a number of analytes or the derivatives thereof in a liquid sample, consisting in the application of the sample to the charging zone (5) of a membrane (2) of the device according to one of cited claims 1 to 8, whereby this sample is present in an amount sufficient for causing the sample liquid to flow through the indicator zones toward the absorption area (3), and for causing the analytes or the derivatives thereof in the sample liquid to form a complex in the indicator zones.

Description

DEVICE AND METHOD FOR SIMULTANEOUSLY IDENTIFYING ANTIGENS OF BLOOD GROUPS DESCRIPTION OF THE INVENTION The invention relates to a device for testing multiple lateral-diagonal flow parameters, particularly in the fields of blood group serology, for the simultaneous qualitative determination and quantitative of several analytes in a liquid sample, comprising a membrane with a loading zone for applying the liquid sample, at least two indicator zones that can enter into a reciprocal reaction with the analyte (s) and at least one absorption region that absorbs the liquid after passing the indicator area, where the indicator areas are located between the loading zone and an absorption zone, characterized in that the direction of flow of the cargo area through the respective indicator zones to an area Absorption (flow paths) are essentially parallel and that there are at least two flow paths different The invention also relates to a method for determining several analytes in a liquid sample comprising the application of the sample in the loading zone of a membrane of the inventive device, this sample being present in a sufficient volume to make the sample liquid flows in the direction of the absorption region through the indicator zones and to cause the analytes or their derivatives in the sample liquid to form a complex, in particular for the simultaneous determination of blood group antigens. In the serological diagnosis of blood groups are generally detected parameters that are important in particular in relation to transfusions respectively hemolytic disease of the newborn. This refers to, among other factors, the detection of antigens on the surface of erythrocytes that are characteristic for blood groups. Other important antigenic systems are also found in thrombocytes, granulocytes, lymphocytes, which also have a role in transfusion and transplantation. As it is known, the erythrocytes of the persons that must be analyzed (donor or receptor) with reagents containing specific blood group antibodies are put together for the determination of blood group antigens. They are usually analyzes of liquids in which a load of analysis is produced by mixing a sample containing erythrocytes that are directed against a certain characteristic of blood group. The analysis load is then incubated for a defined period of time and verified in a defined centrifuge stage either visually or with optical methods with respect to the presence of eventual agglutination or erythrocyte adsorption. The predominant endpoint measurement in blood group serology continues to be haemagglutination. For each blood group, which must be determined, pipette a separate preparation, that is, the determination - for example - of the 9 most important blood groups, A, B, D, C, C, E, Cw and K, requires - without control - 9 separate preparations. Lateral flow assays are often used today as rapid tests, for example as a pregnancy test, to detect markers of infections or as a drug filter. A lateral flow analysis arrangement consists, as is known, of a solid carrier in which it finds a loading zone for the sample to be analyzed, a separation membrane in which ligament elements are bound, for example, respectively captive antigens. and in which ligand reactions can be verified, and an absorptive region capable of absorbing that allows the flow of the sample to be analyzed through the separation membrane. Membranes of analysis for conventional lateral flow analysis are usually described with a separation similar to chromatography. The analyte in the sample binds specifically to a binding element fixed in the membrane, being that these are usually in bands arranged one after the other respectively on top of each other as an indicator area. The ligand complex is visualized by indicator particles that are normally already present in the array in a dried form in a conjugate release pad. The conjugate release pad is located between the loading zone and the membrane. The precoated color indicator particles are coated, for example, with an antibody directed against the target analyte. The usual lateral flow analysis format has the form of a so-called "sandwich pattern" in which both the indicator zone and the indicator particles are loaded with ligands directed against the target analyte, usually an antibody. The ligand (ligament element) is normally held immobile in the membrane. The detection reagent, typically an antibody that is ligated with a particle stained with polystyrene or colloidal metals, is deposited in a washable form on the conjugate release pad. This league complex serves as an indicator particle. The application of the sample to be analyzed rapidly humidifies the conjugate release pad, which mobilizes the indicator particles. The indicator particles migrate with the liquid front along the porous membrane. An analyte found in the sample is bound by the antibody that is coupled to the indicator particle. When the sample passes through the indicator zone, the analyte / indicator particle complex is immobilized in the indicator zone by reaction of the analyte with the antibody bound in the indicator zone, which produces a visible signal. Another known test format for small analytes that have only a single antigenic determinant, which can not simultaneously bind two antibodies, is the so-called "competitive analysis". The detector reagent bound to the indicator particle is normally a molecule identical or analogous to the analyte. The indicator particles are deposited in the conjugate release pad. The indicator particles cross along with the liquid front along the porous membrane. When the sample containing the analyte and the indicator particles (which effectively also contain the analyte) pass through the indicator zone, a part of the analyte molecules binds with the sample and a part with the indicator particles. The greater the amount of analyte found in the sample, the more effectively it will compete with the union of the indicator particles, the weaker the signal becomes. As is known, these indicator particles are for the most part colloidal gold or polystyrene, which are produced and coated by methods known to the expert. In typical lateral flow analysis formats, analytes are detected indirectly. By direct detection of an analyte it is understood here that the analyte is already bound naturally in the indicator particle (e.g., an erythrocyte). In the most usual case of the indirect detection of the analyte, the sample to be analyzed usually contains a non-bound component in cellular form, for example plasma, as an analyte and, in addition to the sample to be analyzed, two reactive components are required, namely a particle indicator and a league element. In indirect detection, the analyte binds first with the reporter particle released from the conjugate release pad, before this complex is immobilized by a second reaction with the linker element in the indicator areas. When conventional lateral flow analyzes with erythrocytes are used as indicator particles that have bound the analytes to be detected, for example blood group-specific antigens, then antibodies against the corresponding blood group antigens in the indicator areas as elements are now available. of league in bands one after another respectively one on top of the other, such as for example anti-A, anti-B against antigens of blood groups A respectively B or antibodies against antigens of the Rh blood group system. For this, conventional lateral flow analyzes have the disadvantage that the erythrocytes bound by the antibodies form, in an analysis, a barrier to the flow of the analytes that still have to be analyzed, for example with respect to other cell-associated antigens. Due to the agglutination or adsorption of cells in a band of ligament elements having a proximal location relative to the loading zone, it is not possible for the other analytes, in particular cells respectively fragments of cells in the sample to be analyzed, to follow their separation without inhibition and visibly and consequently can not be detected unambiguously respectively complete. This can have the consequence, for example, in the case of a person who is of blood group AB Rh D positive, who weakens respectively eliminate band B and D, which could produce a misinterpretation in the sense of blood group A Rh negative Therefore, it has not been possible, until now, to use lateral flow analysis, especially in the serological diagnosis of blood groups, which have more than one indicator area. For the measurement of several parameters of blood groups, in particular linked to cells or of a plasma nature, it is necessary, up to now, to carry out individual parameter analysis separately. The object of the invention is to overcome the aforementioned disadvantages in relation to the previous technology, in particular those of the analysis of respectively detection indicator zones located one after another, one above the other of the conventional lateral flow analyzes, for the measurement simulation of different sample parameters, in particular of cellular and plasma parameters. The objective is achieved inventively by means of a device for the simultaneous qualitative and quantitative determination of one or several analytes in a liquid sample or in several liquid samples, comprising a membrane with a loading zone for applying the liquid sample, at least two zones of indicator that can enter into reciprocal reaction with the analytes and an absorption region that absorbs the liquid after passing through the indicator zones, the indicator areas being located between the loading zone and an absorption region, characterized in that the direction of flow of the loading zone, through the respective indicator zones to an absorption region representing flow tracks is essentially parallel and because there are at least two different flow paths.

The indicator areas of the inventive device are located in the membrane and comprise elements of intercept link respectively bind the analyte (s) in the sample (s) to be detected (s). In the indicator areas, the ligand reactions between analyte and ligand element are detected. In one embodiment of the invention, the indicator areas are arranged in such a way that the sample liquid per flow path passes through only one indicator area. By way of example, the indicator areas are arranged in the membrane in a displaced manner. The arrangement of the indicator areas is configured, in this case, preferably extending diagonally in a row from proximal to distal or in the reverse direction. Special modes are in the form of V, W, M or N, or in the reverse direction of V, W, M or N. In another embodiment, the indicator zones are arranged in a linear row parallelly displaced one next to the other. The introduction of displaced indicator areas is what first allows an analysis of multiple parameters with erythrocytes as indicator particles. The particularly preferred embodiment of a diagonal arrangement has the advantage that the designation of the results can be applied in the inventive arrangement in a particularly practical and easy to read manner, since each of the parameters to be determined has a position X and Y defined, if the arrangement of the inventive device is considered as a coordinate system with ordinate (plane of flow direction) and abscissa (plane of the loading zone). The indicator zones comprise antibodies respectively fragments of antibodies and / or lectins respectively fragments of these which intercept respectively bind the blood group antigens to be determined and with this the cells that carry them. Preferred ligand elements are antibody fragments and / or lectins, respectively fragments of these with the antigens of all blood group systems imaginable in the indicator areas, respectively in the porous membrane. Preferably, a control link element (control = ctl), which indicates positively, is applied in an indicator area of this row of indicator areas, preferably in a zone of indicator of distal location with respect to all other indicator areas. the passage of the sample through the indicator areas. The control ligand element is preferably a polyclonal anti-erythrocyte antibody. In a preferred embodiment, respectively, an indicator zone comprises in this a ligament element, preferably an antibody respectively an antibody fragment against an analyte to be analyzed. Preferred moieties of antibodies respectively of antibody fragments and / or lectins respectively fragments thereof in the indicator zones are respectively lectin antibodies against antigens of the ABO blood group system, Rh system, Kell, Lewis, Hh, Duffy, Kidd, MNS , Lutheran, P. Also preferred as ligand elements in the indicator zones are antibodies against antigens of blood group systems Diego, Yt, Scianna, Dombrock, Colton, Chido / Rodgers, Gerbich, Cromer, Knops, Landsteiner-Wiener, Xg, Kx, Indian, Ok, Raph, John Milton Hagen, Langereis and / or Sid. A particularly preferred embodiment of the inventive device comprises indicator areas with ligand elements anti-A, -B, -AB, -D, D antibodies , -C, -c, -E, -e, -Cw and / or K respectively their antibody fragments, being that the two anti-D antibodies are two different antibodies, respectively the antibody fragments of these. In particular in the case of patients, pregnant or neonates, these are preferably monoclonal antibodies of the IgM class that do not detect the DVI category. In the case of donors, it is preferably an antibody that detects the DVI category and an antibody that does not detect the DVI category.

Thanks to the inventive device, it is no longer necessary to pipette separately for each individual determination, but a large number of antigens desired for blood group systems to be analyzed can be determined in a sample simultaneously at a time, for example the most important characteristics of the systems of blood groups ABO, Rh and Kell (A, B, AB, D, C, c, E, e, Cw, K). This represents an extraordinary rationalization of work procedures. Also the reading of the results represented in diagonal arrangement is substantially more favorable. Furthermore, it is possible to determine and read in the inventive device, for example, properties of ABO and Rh in a single device, one next to the other. The association of the results with the respective patient is facilitated. The two-dimensional result, as well as the stable end point of the reaction, favors both the reading to the naked eye and an automated reading of the results with the usual methods of graphic analysis, such as CCD cameras. The work investment is reduced even in the case of manual processing. The inventive device also produces a reduction of environmental impact and favorable effects on costs. Even in emergency situations with time pressure it can be carried out in a short time in a single step of analysis, for example, a complete determination of ABO blood groups / Rh subgroups. Regarding the production technique, the lateral-diagonal flow construction offers substantial advantages in relation to the previous technology, being that there is a substantially lower consumption of reagents used and the offering of a multiplicity of analysis parameters in a single device. Thanks to the inventive device, a lateral flow analysis is offered, in particular for the serological diagnosis of blood groups, in which erythrocytes are used as indicator particles and with which a plurality of plasmatic parameters and / or cell characteristics are allowed simultaneously in a sample preparation. blood, in particular antigens of erythrocytes respectively epitopes of antigens, in particular of components of whole blood, in each sample to be analyzed. Additionally, an analysis system that is easy to produce and economical and easy to use is offered in this way, particularly with few series of tests and without sample preparation, which allows the simultaneous determination of different cellular parameters and / or plasma parameters of a sample, in particular characteristics of blood groups. The membrane of the inventive device is a porous membrane. Preferred materials for the membrane are, for example, nitrocellulose (for example UniSart from Sartorius, HiFlow from Millipore, Whatman, AE99 respectively FF85 / 100 from Schleicher &Schuell), polyethylene. { Lateral Fio from Porex Corporation) or Nylon. { Novylon from CUNO). Preferably, the membrane has a pore size as high as possible, since a high porosity of the membrane facilitates the penetration into the porous structure, in particular, of cellular components of the sample to be analyzed, e.g. of erythrocytes. Particularly advantageous is the use of absorbent membranes. The inventive device, however, is not restricted to these characteristics. All membranes having a high capillary flow rate (Capillary Speed) are preferred, being that the rate of capillary flow is the time that a dyeing solution requires to advance 40 mm in a given membrane. In particular, membranes with a capillary flow rate < 100. In a preferred embodiment of the invention, in the direction of flow behind the loading area of the inventive device, there is a sealing element disposed in the porous membrane. Two- or three-dimensional sealing elements are applied which are placed on the porous membrane and with which a sample loading zone is generated, separated from the rest of the surface of the porous membrane. The sealing element has, firstly, the effect of a liquid barrier and allows the targeted distribution of the sample liquid and the analysis reagents in the porous membrane. In addition, the sealing element inventively seals the sample application area to prevent an undesirable passage of liquid to the other areas of the lateral flow device arrangement. Preferred embodiments of the sealing element are in the form of a rib, respectively a funnel vat. The shaping of the sealing element is carried out by cutting processes of the material used for the production of the sealing element. In the case of the funnel or tub shape, the sealing element comprises an internal opening whose preferred variants of modalities are round, square or rectangular shapes, in the case of a funnel shape, decreasing towards the lower side (contact side). of the membrane) of the sealing element. Preferred materials for the sealing element are materials that do not absorb water (hydrophobic) In a particular embodiment, the materials are coated unilaterally with an adhesive film, for example a pressure-sensitive acrylate adhesive respectively self-adhesive. In this way, the sealing element can be stuck directly on the surface of the porous membrane. As an alternative, the sealing element can be joined to the lateral flow box, for example, bonded, wherein in this embodiment the lateral flow box presses the sealing element onto the surface of the porous membrane, thus establishing the operation of the sealing element. Preferred materials for the formation of two-dimensional sealing elements are any form of adhesive tapes or adhesive sheets (for example, Tesa 4124 of Beiersdorf AG, ARcare 7815 of Adhesive Research). Preferred materials for the formation of three-dimensional sealing elements are flexible elastomer materials with closed pores or flexible silicone materials with different thicknesses of material, preferably 3-5 mm (for example, Pitzner EPDM140 cellular rubber, silicone rubber or whole rubber) , hardness 40 ° or less, from Castan). Thanks to this inventive conditioning, the inventive device can receive liquid samples containing cells, such as, for example, whole blood, without separating the cells in the process. In addition, the sealing element allows the application of large volumes of sample in the porous membrane (loading zone) without it flooding. The sealing element thus helps to exploit the absorbent properties of the porous membrane. In addition, the sealing element guarantees a directed flow of sample. The inventive device can, however, function well with or without a sealing element. For the absorption region (absorption pad) of the inventive device, mechanically stable materials are preferable, preferably with water absorption capacities of 20-30 g / 100 cm 2 (for example Wicking paper, type 300, Schleicher und Schüll). The contact between the absorption pad and the lateral flow membrane of the inventive device is produced by press-contact or overlap with the porous membrane. The precise placement of the absorption pad in the membrane is achieved by gluing the absorption pad with the carrier layer that carries the lateral flow membrane (backing sheet). In another embodiment, the components of the inventive device are fixed on a support or a carrier layer respectively for the purpose of providing mechanical reinforcement. But the inventive device can work with or without a carrier layer. Mechanically stable and non-water-absorbing materials are preferred, preferably with material thicknesses of 100 μm or more, which are coated on one or both sides with an adhesion film, for example a pressure-sensitive acrylate adhesive respectively self-adhesive (e.g. , 0.005"polyester W / GL-187, G &L) .The porous membrane and the absorption pad are fixed to the carrier layer.In the case of an adherent carrier layer on both sides, the second adherent face is used to fixing the arrangement on other surfaces, for example inside the lateral flow box In another embodiment, the inventive device is integrated with or without a carrier layer, in which the components of the inventive device are fixed, in a box, which presses the components of the membrane against each other and the box thus supports the function of sealing element.The inventive device can work, however, even l of good with or without box. Another object of the invention is the use of the inventive device for blood analysis, in particular for the simultaneous determination of antigens of blood groups or epitopes of antigens of all imaginable blood group systems, preferably of any analyte on the surface of the erythrocytes. The antigens • respectively antitope epitopes to be analyzed are, by way of example, those of the ABO blood group system, of the Rh, Kell, Lewis, Hh, Duffy, Kidd, MNS, Lutheran, P. systems of the Diego blood group systems , Yt, Scianna, Dombrock, Colton, Chido / Rodgers, Gerbich, Cromer, Knops, Landsteiner-Wiener, Xg, Kx, Indian, Ok, Raph, John Milton Hagen, Langereis and / or Sid, in particular Al, A2, B , D, C, c, E, e, Cw, K, k, M, N, S, s, Jk (a), Jk (b), Fy (a), Fy (b), Kp (a), Kp (b), Js (a), Js (b), Le (a), Le (b), Lu (b), Pl, I, H, Xg (a), 'U, Vw, Wr (a) , Lan. A particularly preferred embodiment of the inventive device simultaneously determines several characteristics of blood groups, by way of example A, B, AB, D, C, c, E, e, Cw and K. The samples to be analyzed, for example native whole blood or Anti-coagulated or concentrated erythrocytes or suspensions of diluted erythrocytes, is applied in the loading area of the inventive device. The erythrocytes contained in the sample carrying the analyte (s) simultaneously serve as indicator particles. The objective is achieved, on the other hand, by a method for the determination of several analytes or their derivatives in a liquid sample comprising the application of the sample in the loading zone of a membrane of the inventive device, this sample being present in a sufficient volume to cause the sample liquid to flow in the direction of the absorption region through the indicator zones and to cause the analytes or their derivatives in the sample liquid to be linked with the respective indicator zones respectively form a complex in the indicator areas. In the inventive method, the analytes to be determined are in particular antigens of blood groups or epitopes of antigens of all blood group systems, preferably of those found on the surface of erythrocytes. The antigens respectively epitopes of antigens are, by way of example, that of the ABO blood group system, of the Rh, Kell, Lewis, Hh, Duffy, Kidd, MNS, Lutheran, P. systems of the Diego blood group systems, Yt, Scianna, Dombrock, Colton, Chido / Rodgers, Gerbich, Cromer, Knops, Landsteiner-Wiener, Xg, Kx, Indian, Ok, Raph, John Milton Hagen, Langereis and / or Sid, in particular Al, A2, B, D, C, c, E, e, Cw, K, k, M, N, S, s, Jk (a), Jk (b), Fy (a), Fy (b), Kp (a), Kp (b), Js (a), Js (b), Le (a), Le (b), Lu (b), Pl, I, H, Xg (a), U, Vw, Wr (a), Lan . A particularly preferred embodiment of the inventive method simultaneously determines several characteristics of blood groups, by way of example A, B, AB, D, C, c, E, e, Cw and K. The samples to be analyzed, for example, native whole blood or Anti-coagulated or concentrated erythrocytes or suspensions of diluted erythrocytes, is applied in the loading area of the inventive device. The erythrocytes contained in the sample carrying the analyte (s) simultaneously serve as indicator particles. The invention is explained below by means of figures and examples in more detail, without limiting them. In the figures there is shown: Fig. 1 a perspective representation of an inventive device for lateral flow analysis for the simultaneous determination of the characteristics of blood groups A, B, AB, D and CDE; FIG. 2 is an exploded view of the inventive device for lateral flow analysis, shown in FIG. 1; FIG. 3 a perspective representation of an inventive device for lateral flow analysis for the simultaneous determination of the characteristics of blood groups A, B, AB, D and CDE, made with a three-dimensional obturation element in the form of a rib; Fig. 4 an exploded representation of the inventive device for lateral flow analysis, shown in Fig. 3; Fig. 5 a perspective representation of an inventive device for lateral flow analysis for the simultaneous determination of the characteristics of blood groups A, B, AB, D and CDE, made with a tubular three-dimensional sealing element; Fig. 6 an exploded view of the inventive device for lateral flow analysis, shown in Fig. 5; FIG. 7 a perspective representation of an inventive device for lateral flow analysis for the simultaneous determination of the characteristics of blood groups A, B, AB, D, C, c, E, e, Cw and K; Fig. 8 a perspective representation of an inventive device for lateral flow analysis performed as bedside analysis for the verification of receptor identity and conserve. Fig. 9 an exploded representation of the inventive device shown in Fig. 8 for lateral flow analysis. FIG. 10 a perspective representation of an inventive device for lateral flow analysis for the simultaneous determination of the characteristics of blood groups A, B, AB, D and CDE. Fig. 11 a perspective representation of an inventive device for lateral flow analysis for the simultaneous determination of the characteristics of blood groups A, B, AB, D and CDE.

Fig. 12 a perspective representation of an inventive device for lateral flow analysis for the simultaneous determination of the characteristics of blood groups A, B, AB, D and CDE. Fig. 13 a perspective representation of an inventive device for lateral flow analysis for the simultaneous determination of the characteristics of blood groups A, B, AB, D and DCE. Fig. 14 a perspective representation of an inventive device for lateral flow analysis with bidirectional flow for the simultaneous determination of the characteristics of blood groups A, B, AB, D and DCE. Fig. 15 an exploded view of the inventive device for lateral flow analysis shown in Fig. 14. In Fig. 1 there is shown, by way of example, a perspective representation of an inventive device for lateral flow analysis for analysis of lateral flow for the simultaneous determination of the characteristics of blood groups A, B, AB, D and CDE. In the present example, the device consists of a carrier layer 1, of the porous membrane 2, the absorption pad 3 and the two-dimensional sealing element 4, realized in the form of a rib. The porous membrane 2 is fixed, in this case, in the carrier layer 1 provided with a pressure sensitive acrylate adhesive. The absorption pad 3 flaps, on the other hand, with the porous membrane 2. The sealing element 4 fixed on the upper face of the porous membrane 2 separates the loading zone 5 from the rest of the membrane surface and allows the targeted distribution of analysis liquid and analysis reagents in the porous membrane 2. Between the loading zone 5 and the region of the porous membrane 2 which makes contact with the absorption pad 3, the region 6 of indicator areas is arranged. This is formed by zones I-VI of indicator point, displaced diagonally, arranged in defined X and Y positions, being that the indicator areas consist of the following elements of league: areas of indicator element of league specification I antibodies anti-A (monoclonal) II antibodies anti-B (monoclonal) III antibodies anti-AB (monoclonal) IV antibodies anti-D (monoclonal) V anti-CDE antibodies (monoclonal) VI anti-erythrocyte antibodies (polyclonal) Indicator zone VI is the control (ctl) and contains anti-polyclonal erythrocyte antibodies. It is located distally in relation to all other indicator areas. In Fig. 2 an exploded representation of the inventive device for lateral flow analysis shown in Fig. 1, consisting of the carrier layer 1 components, porous membrane 2, absorption pad 3 and sealing element 4 separating the area, is shown. 5 of the rest of the membrane, which in turn consists of the region 6 of indicator areas, with the indicator areas I-VI arranged diagonally displaced. In Fig. 3 there is shown, by way of example, a perspective representation of an inventive device for lateral flow analysis for the simultaneous determination of the characteristics of blood groups A, B, AB, D and DCE. In the present example, the components of the device correspond to the components of the device shown in Fig. 1, with the exception of the sealing element 4 made in the form of a three-dimensional rib, fixed on the upper face of the porous membrane 2. In Fig. 4 an exploded representation of the inventive device for lateral flow analysis shown in Fig. 3 is shown with the carrier layer 1 components, porous membrane 2, absorption pad 3 and sealing element 4 made in the form of a three-dimensional rib, which separates the load zone 5 from the rest of the membrane, which again contains the region 6 of the indicator zones with the I-VI indicator areas arranged in a diagonally offset manner. Fig. 5 shows, by way of example, a perspective representation of an inventive device for lateral flow analysis for the simultaneous determination of the characteristics of blood groups A, B, AB, D and DCE. In the present example, the components of the device correspond to the components of the device shown in Fig. 1, with the exception of the sealing element 4 made in the form of a three-dimensional tub, fixed on the upper face of the porous membrane 2. In Fig. 6 an exploded representation of the inventive device for lateral flow analysis shown in Fig. 5 is shown with the carrier layer 1 components, porous membrane 2, absorption pad 3 and sealing element 4 made in the form of a three-dimensional tub, which separates the load zone 5 from the rest of the membrane, which again contains the region 6 of the indicator zones with the I-VI indicator areas arranged in a diagonally offset manner. In Fig. 7 there is shown, by way of example, a perspective representation of an inventive device for the simultaneous determination of the characteristics of blood groups A, B, AB, DC, c, E, e, Cw and K. In the present example, the device consists of a carrier layer 1, the porous membrane 2, the absorption pad 3 and the bi-axial sealing element 4 formed in the form of a rib. The porous membrane 2 is fixed, in this case, in the carrier layer 1 provided with a pressure sensitive acrylate adhesive. Also the absorption pad 3 is fixed in the carrier layer 1, with a part of the absorption pad 3 overlapping the porous membrane 2. The sealing element 4, fixed on the upper face of the porous membrane 2, separates the loading zone 5 from the rest of the membrane surface and allows a directed distribution of sample liquid and analysis reagents to the porous membrane 2. Between the loading zone 5 and the region of the porous membrane 2 which makes contact with the absorption pad 3, the region 6 of indicator areas is arranged. This is formed by punctiform I-XI zones, the indicator zones consist of the following ligand elements: indicator zones, ligand element, specification I, anti-A antibodies (monoclonal), II, anti-B (monoclonal) antibodies, III, anti-AB antibodies. (monoclonal) IV anti-D (monoclonal) antibodies V anti-C (monoclonal) antibodies VI anti-c (monoclonal) antibodies VII anti-E (monoclonal) antibodies VIII anti-e antibodies (monoclonal) IX anti-Cw antibodies (monoclonal ) X anti-K (monoclonal) antibodies XI anti-erythrocyte (polyclonal) antibodies Zone XI of the indicator is the control (ctl) and contains antibodies against anti-polyclonal erythrocytes. FIG. 8 shows, by way of example, a perspective representation of an inventive device for lateral flow analysis carried out as analysis next to the bed to verify the ABO identity of the receiver and conserve. In the present example, the device consists of a carrier layer 1, of the porous membrane 2a and 2b present in double materialization, the absorption pad 3 and the two-dimensional sealing element 4 realized in the form of a rib. The two porous membranes 2a and 2b are fixed to the carrier layer 1 provided with a respectively self-adhesive pressure-sensitive acrylate adhesive. Also the absorption pad 3 is fixed in the carrier layer 1, a part of the absorption pad 3 overlapping both porous membranes 2 at the same distance. The sealing elements 4a and 4b, fixed on the upper faces of the porous membranes 2a and 2b, separate the respective charging zones 5a and 5b from the rest of the membrane surface and allow a directed distribution of sample liquid and analysis reagents. to porous membranes 2. Between the charging zones 5a respectively 5b and the respective regions of the porous membranes 2a and 2b contacting the absorption pad 3, the regions 6a and 6b of indicator areas are arranged. These are formed by punctiform zones Ia-IIIa respectively Ib-IIIb, displaced diagonally, arranged in defined X and Y positions, where the indicator areas consist of the following elements of the league: indicator areas element of the league specification la, Ib antibodies anti-A (monoclonal) Ha, Ilb antibodies anti-B (monoclonal) Illa, Illb antibodies anti-AB (monoclonal) Illa and Illb areas of indicator are the controls (ctl) and contain polyclonal anti-erythrocyte antibodies . It is distal to all other indicator areas. In Fig. 9 there is shown an exploded representation of the inventive device for lateral flow analysis shown in Fig. 8 with the carrier layer 1 components, porous membranes 2a and 2b, absorption pad 3 and the sealing elements 4a and 4b that separate , respectively, the loading areas 5a and 5b of the rest of the membrane, which itself contains the regions 6a respectively 6b of indicator areas comprising the indicator areas I-1 and Ib-Illb arranged diagonally from proximal to distal . FIG. 10 shows, by way of example, a perspective representation of an inventive device for lateral flow tests for the simultaneous determination of the characteristics of blood groups A, B, AB, D and DCE. The present example represents a lateral flow analysis device for right-handers and consists of a carrier layer 1, the porous membrane 2, the absorption pad 3 and the two-dimensional sealing element 4 realized in the form of a rib. The porous membrane 2 is fixed, in this case, in the carrier layer 1 provided with a pressure sensitive acrylate adhesive. Also the absorption pad 3 is fixed in the carrier layer 1, wherein a part of the absorption pad 3 overlaps the porous membrane 2. The sealing element 4, fixed on the upper face of the porous membrane 2, separates the loading zone 5 from the rest of the membrane surface and allows a directed distribution of sample liquid and analysis reagents to the porous membrane 2. Between the loading zone 5 and the region of the porous membrane 2 which makes contact with the absorption pad 3, the region 6 of indicator areas is arranged. This is formed by punctiform zones I-VI, displaced in parallel one next to the other, arranged in a linear row in defined X and Y positions, being that the indicator zones consist of the following elements of league: indicator areas element of league specification I antibodies anti-A (monoclonal) II antibodies anti-B (monoclonal) III antibodies anti-AB (monoclonal) IV antibodies anti-D (monoclonal) V antibodies anti-CDE (monoclonal) VI antibodies anti-erythrocytes (polyclonal) Zone VI of indicator is the control (ctl) and contains anti-erythrocyte antibodies. It is distal to all other indicator areas. In Fig. 11 there is shown, by way of example, a perspective representation of an inventive device for lateral flow tests for the simultaneous determination of the characteristics of blood groups A, B, AB, D and DCE. The present example represents a lateral flow analysis device for lefties and consists of a carrier layer 1, the porous membrane 2, the absorption pad 3 and the two-dimensional sealing element 4 made in the form of a rib. The porous membrane 2 is fixed, in this case, in the carrier layer 1 provided with a pressure sensitive acrylate adhesive. Also the absorption pad 3 is fixed in the carrier layer 1, wherein a part of the absorption pad 3 overlaps the porous membrane 2. The sealing element 4, fixed on the upper face of the porous membrane 2, separates the loading zone 5 from the rest of the membrane surface and allows a directed distribution of sample liquid and analysis reagents to the porous membrane 2. Between the loading zone 5 and the region of the porous membrane 2 which makes contact with the absorption pad 3, the region 6 of indicator areas is arranged. This is formed by punctiform zones I-VI, displaced in parallel one next to the other, arranged in a linear row in defined X and Y positions, being that the indicator zones consist of the following elements of league: indicator areas league element specification I antibodies anti-A (monoclonal) II antibodies anti-B (monoclonal) III antibodies anti-AB (monoclonal) IV antibodies anti-D (monoclonal) V antibodies anti-CDE (monoclonal) VI antibodies anti-erythrocytes (polyclonal) Zone VI of indicator is the control (ctl) and contains anti-polyclonal erythrocyte antibodies. It is distal to all other indicator areas. FIG. 12 shows, by way of example, a perspective representation of an inventive device for lateral flow tests for the simultaneous determination of the characteristics of blood groups A, B, AB, D and DCE. The present example represents a lateral flow analysis device for right-handers and consists of a carrier layer 1, the porous membrane 2, the absorption pad 3 and the two-dimensional sealing element 4 realized in the form of a rib. The porous membrane 2 is fixed, in this case, in the carrier layer 1 provided with a pressure sensitive acrylate adhesive. Also the absorption pad 3 is fixed in the carrier layer 1, with a part of the absorption pad 3 overlapping the porous membrane 2. The sealing element 4, fixed on the upper face of the porous membrane 2, separates the loading zone 5 from the rest of the membrane surface and allows a directed distribution of sample liquid and analysis reagents to the porous membrane 2. Between the loading zone 5 and the region of the porous membrane 2 which makes contact with the absorption pad 3, the region 6 of indicator areas is arranged. This is formed by zones I-VI stretched longitudinally respectively in the form of tapes, displaced in parallel one next to the other, arranged in defined X and Y positions, where the indicator areas consist of the following elements of ligament: of indicator element of league specification I antibodies anti-A (monoclonal) II antibodies anti-B (monoclonal) III antibodies anti-AB (monoclonal) IV antibodies anti-D (monoclonal) V antibodies anti-CDE (monoclonal) VI antibodies anti-erythrocytes (polyclonal) Zone VI of indicator is the control (ctl) and contains polyclonal anti-erythrocyte antibodies. It is distal to all other indicator areas. FIG. 13 shows, by way of example, a perspective representation of an inventive device for lateral flow tests for the simultaneous determination of the characteristics of blood groups A, B, AB, D and DCE. The present example represents a lateral flow analysis device for lefties and consists of a carrier layer 1, the porous membrane 2, the absorption pad 3 and the two-dimensional sealing element 4 made in the form of a rib. The porous membrane 2 is fixed, in this case, in the carrier layer 1 provided with a pressure sensitive acrylate adhesive. Also the absorption pad 3 is fixed in the carrier layer 1, with a part of the absorption pad 3 overlapping the porous membrane 2. The sealing element 4, fixed on the upper face of the porous membrane 2, separates the loading zone 5 from the rest of the membrane surface and allows a directed distribution of sample liquid and analysis reagents to the porous membrane 2. Between the loading zone 5 and the region of the porous membrane 2 which makes contact with the absorption pad 3, the region 6 of indicator areas is arranged. This is formed by zones I-VI stretched longitudinally respectively in the form of tapes, displaced in parallel one next to the other, arranged in defined X and Y positions, where the indicator areas consist of the following elements of link: indicator areas league element specification I antibodies anti-A (monoclonal) II antibodies anti-B (monoclonal) III antibodies anti-AB (monoclonal) IV antibodies anti-D (monoclonal) V anti-CDE antibodies (monoclonal) VI anti-erythrocyte antibodies (polyclonal) Indicator zone VI is the control (ctl) and contains polyclonal anti-erythrocyte antibodies. It is distal to all other indicator areas. FIG. 14 shows, by way of example, a perspective representation of an inventive device for lateral flow tests with bidirectional flow for the simultaneous determination of the characteristics of blood groups A, B, AB, D and DCE. The present example consists of a carrier layer 1, the porous membrane 2, the absorption pads 3a and 3b and the two-dimensional obturation elements 4a and 4b made in the form of a rib. The porous membrane 2 is fixed, in this case, in the carrier layer 1 provided with a pressure sensitive acrylate adhesive. Also the absorption pads 3a and 3b are fixed in the carrier layer 1, with a part of the absorption pads 3a and 3b overlapping the porous membrane 2. The sealing elements 4a and 4b, fixed on the upper face of the porous membrane 2, separate the loading zone 5 from the rest of the membrane surface and allow a directed distribution of sample liquid and analysis reagents to the porous membrane 2 . Between the charging zone 5 and the region of the porous membrane 2 which contacts the absorption pads 3a and 3b, the regions 6a and 6b of indicator areas are arranged. These are formed by punctiform I-VI zones, displaced diagonally, arranged in defined X and Y positions, being that the indicator zones consist of the following elements of the league: indicator areas element of the league specification I antibodies anti-A (monoclonal) II antibodies anti-B (monoclonal) III antibodies anti-AB (monoclonal) IV antibodies anti-D (monoclonal) V antibodies anti-CDE (monoclonal) VIa, VIb antibodies anti-erythrocytes (polyclonal) The Vía or VIb indicator zones are the controls (ctl) and contain polyclonal anti-erythrocyte antibodies. They are located distally in relation to all zones I-III respectively indicator IV-V. In FIG. 15, the inventive device shown in FIG. 14 for lateral flow analysis with bidirectional flow consisting of a carrier layer 1, the porous membrane 2, the absorption pads 3a and 3b and the elements is shown in an exploded view. 4a and 4b of sealing that separate the load zone 5 located in the center of the rest of the membrane surface again contain the two regions 6a and 6b of zones I, II, III, Via respectively, IV, V, VIb of indicator, displaced diagonally from proximal to distal. Examples Example 1: Blood group determination Preparation of the test strips The test strips consist of a loading zone, a region of indicator zones and an absorption region. Membranes of the Millipore HiFlow Plus 065 type are cut to form strips with a size of 15 x 35 mm (width / length, x / y) and are stuck in a carrier layer (Backing sheet, for example G &L). They are applied in the region of diagonally displaced indicator 'areas, a number of respectively 0.2 μl with the aid of a dispenser, for example AD 3200 (Biodot): Anti-A-clone Birma-1 (Serologicals, TLJ0105); anti-B-clone ES-4 (Serologicals, NCA0201); anti-AB-clones AB6, AB26, AB92 (Medion Diagnostics, 010062); anti-D-clone MS-80 + MS-258 (Serologicals, KXE0201); anti-C -clone MS-24 (Serologicals, not formulated, KGK0212) M; anti-c -clone MS-33 (Serologicals, KNI0207); anti-E-clone MS-80 + MS-258 (Serologicals, KXE0201); anti-e clones MS-21 + MS-63 (Serologicals, KLL0205 + KQK0205); anti-Cw-clone MS-110 (Serologicals, JPK0201); anti-K clone MS-56, (Serologicals KOA0201). The positioning of anti-A antibody is carried out in position x = 3 mm / y = 10 mm. All other antibodies are dispensed iteratively at distances of x = 1.5 mm / y = 2.2 mm relative to the position of the anti-A antibody. The anti-erythrocyte specific control antibody (Rabbit IgG anti human RBC fraction, Rockland, 209-4139) is applied at x = 2 / y = 3.5 mm displacement in relation to the last point of the series of antibodies specific for blood groups. Antibody dilution is performed in 15 M potassium phosphate buffer, pH 7.5, 10% (v / v) methanol) as follows: anti-A antibody 1: 3, anti-B antibody 1: 2, anti-antibody AB 1: 4, anti-D antibody 1: 4, anti-RBC antibody 1: 3. All other antibody solutions are not pre-diluted, but mixed with 10% (v / v) methanol. The membranes are dried, after dispensing the antibodies, for 20 min at 40 ° C and then stored under constant atmospheric humidity until analysis is carried out. At the distal end in relation to the loading area an absorption pad is adhered, which overlaps the membrane by 3 mm, measuring 15 x 10 mm respectively 26 x 10 mm (Schleicher & amp;; Schüll, 300). The loading zone is separated from the rest of the membrane by adhesion, over the entire membrane width, of an adherent strip with a width of 1-2 mm (Tesa 4124) in position y = 5 mm. Analysis load: Anticoagulated whole blood is used as blood samples. For the analysis itself 100 μl of blood diluted 1: 6 in dilution buffer (EnlisstII, Medion Diagnost cs or Diluent 1, DiaMed) are applied in loading zone (6-point mode) respectively 150 μl (11-point mode) ). When the blood has left the loading zone, 100 μl respectively 150 μl of buffer to dilute or preferably with 100 μl of hypo-osmotic wash buffer (15 mM potassium phosphate buffer, pH 7.4, 0.3-0.45) is applied. % (v / v) NaCl) to remove unbound erythrocytes from the membrane. Alternatively, the sample application can also be performed with 50 μl of diluted or undiluted blood. In these analyzes the membrane is washed twice with buffer to dilute respectively once with buffer to dilute followed by hypo-osmotic wash buffer. With the dilution selected, the anti-RBC control becomes visible after 2 minutes as an indicator of a successful analysis. The analysis is slower with undiluted blood. Result: The analysis is valid if the anti-RBC control shows a clearly positive signal (red dot). Depending on the presence or absence of the respective blood group antigens, red spots appear in the corresponding positions (positive) or the almost white background color of the membrane (negative). Example 2: analysis next to the bed Preparation of the analysis strips: The analysis next to the bed consists of two membranes respectively ("conserve", "receptor") fixed in a carrier layer (support sheet) consisting respectively of a loading zone, a region of indicator zones and an absorption region . Membranes of the Millipore HiFlow Plus 065 type are cut to form strips with a size of 12.5 x 30 mm • (width / length; x / y). Two of these are respectively glued to a carrier layer (Backing sheet, for example G &L) at a distance of 5 mm, so that the total size of the assembly is 30 x 30 mm. They are applied in both membranes with the help of a dispenser, for example AD 3200 (Biodot), respectively the same applications: 0.2 μl of solutions of monoclonal antibodies anti-A - clone Birma-1 (Serologicals, TLJ0105) in position x = 4 / y = 12 mm; anti-B - clone ES-4 (Serologicals, NCA0201) in position x = 7 / y = 14 mm. The control antibody specific antieritrocitos (Rabbit IgG Fraction of anti Human RBC, Rockland, 209-4139) is applied with x = 3 / y = 4 mm displacement in relation to the anti-B point. Antibody dilution is performed on the antibodies in 15 mM potassium phosphate buffer, pH 7.5, 10% (v / v) methanol) as follows: anti-A 1: 3 antibody, anti-B 1 antibody: 2, anti-RBC antibody 1: 3.

The membranes are dried, after dispensing the antibodies, for 20 min at 40 ° C and then stored under constant atmospheric humidity until analysis is carried out. At the distal end in relation to the loading zone, an absorption pad is adhered, which overlaps the membrane by 3 mm, measuring 30 x 10 mm (Schleicher &Schüll, 300). The loading zone is separated from the rest of the membrane by adhesion, over the entire membrane width, of an adherent strip with a width of 1-2 mm (Tesa 4124) in position y = 5 mm. Analysis load: As blood samples, it is used: for the "preserves" membrane: erythrocyte concentrate; for the "receptor" membrane: whole blood. For the analysis itself, 50 μl of whole blood is applied to the "receptor" side and 50 μl of erythrocyte concentrate on the "preserved" side in the respective loading zone. When the blood has left the loading zone, it is washed with respectively 2 x 100 μl of buffer to dilute respectively once with buffer to dilute and then with hypo-osmotic wash buffer. Result: The anti-RBC control as an indicator of a successful analysis is visible on both membranes after approximately 2 minutes.

The analysis is valid if the anti-RBC control shows a clearly positive signal (red dot). Depending on the presence or absence of the respective blood group antigens, red spots appear in the corresponding positions (positive) or the almost white background color of the membrane (negative). An identical box for? "receiver" and "conserve" means ABO identity between receiver and conserve. Example 3: Determination of blood group with bidirectional lateral flow analysis Preparation of the analysis strips: The analysis strips consist of a loading zone located in the center, two regions of indicator zones and two absorption regions. Millipore HiFlow Plus 065 type membranes are cut to form strips with a size of 15 x 50 mm (width / length, x / y) and are stuck in a carrier layer (Backing sheet, for example G &L). Shifted diagonally or alternatively displaced in a linear row, 0.2 μl of solutions of different monoclonal antibodies specific for blood groups are applied in the region of indicator areas. Half of the strip length (Y = 0) is in this case the reference quantity for the positioning of the indicator areas in the y-direction. The following antibodies are dispensed using a dispenser, for example, AD3200 (Biodot): Anti-A-clone Birma-1 (Serologicals, TLJ0105); anti-B-clone ES-4 (Serologicals, NCA0201); anti-AB-clones AB6, AB26, AB92 (Medion Diagnostics, 010062); anti-D-clone MS-80 + MS-258 (Serologicals, KXE0201); anti-C -clone MS-24 (Serologicals, non-formulated, KGK0212); anti-E -clone MS-80 + MS-258 (Serologicals, KXE0201). For the anti-CDE indicator zone, the anti-D and anti-C antibodies are concentrated twice and the anti-E antibody three times and mixed in equal parts by volume. In the modality with diagonal displacement of the indicator areas, the anti-A antibody is dispensed in position x = 4 / y = 10 mm. The positions of anti-B and anti-AB antibodies are carried out in iterated form at distances of x = 3.5 / y = 2 mm in relation to the position of anti-A antibody. The anti-erythrocyte specific control antibody (Rabbit IgG Fraction of anti Human RBC, Rockland, 209-4139) is applied with x = 3.5 / y = 3.5 mm displacement relative to the last point of the series of anti-A antibodies, anti-B and anti-AB. The anti-D antibody is dispensed in the x = 4 / y = -10 mm position, the anti-CDE antibody is dispensed at the distance of x = 3.5 / y = -2 mm. The anti-erythrocyte specific control antibody (Rabbit IgG Fraction of anti Human RBC, Rockland, 209-4139) is applied with x = 3.5 / y = -3.5 m displacement relative to the anti-CDE antibody point. Antibody dilution is performed on the antibodies in 15 mM potassium phosphate buffer, pH 7.5, 10% (v / v) methanol) as follows: anti-A 1: 3 antibody, anti-B 1 antibody: 2, anti-RBC antibody 1: 3. The anti-CDE antibody mixture is not pre-diluted, but mixed with 10% (v / v) methanol. The membranes are dried, after dispensing the antibodies, for 20 min at 40 ° C and then kept under constant atmospheric humidity until analysis is carried out. At the distal end in relation to the loading zone, an absorption pad is adhered, which overlaps the membrane by 3 mm, measuring 15 x 10 mm (Schleicher &Schüll, 300). The loading area is separated from the rest of the membrane by adhesion, over the entire membrane width, of an adherent strip with a width of 1-2 mm (Tesa 4124) in position y = 3 mm respectively y = -3 mm. Analysis load: whole blood is used as blood samples. For the analysis itself 100 μl of diluted whole blood is applied to l: 6 in dilute buffer (EnlisstII, Medion Diagnostics) in the respective loading zone. When the blood has left the loading zone, pipette once with 100 μl of buffer to dilute or with 100 μl of hypo-osmotic wash buffer (15 mM potassium phosphate buffer pH 7.4, 0.3-0.45% ( w / v) NaCl) in the loading zone to remove unbound erythrocytes from the membrane. Alternatively, however, sample application can also be performed with 50 μl of blood diluted 1: 3 or undiluted blood. In these samples the membrane is washed twice with buffer to dilute respectively once with buffer to dilute and then with hypo-osmotic buffer to wash. With the dilution of 1: 6 selected, the anti-RBC control becomes visible as an indicator of a successful analysis after 2 minutes. With undiluted blood, the analysis takes more time. Result: The analysis is valid if the anti-RBC control shows a clearly positive signal (red dot). Depending on the presence or absence of the respective blood group antigens, red spots appear in the corresponding positions (positive) or the almost white background color of the membrane (negative).

Claims (19)

CLAIMING

1. Device for the simultaneous, qualitative or quantitative determination of several analytes in a liquid sample, comprising a membrane with - a loading zone for applying a liquid sample, - at least two indicator zones that can enter into reciprocal reaction with the analyte (s) and - at least one absorption zone that absorbs the liquid after passing through the indicator zones, wherein the indicator zones are located between the loading zone and an absorption region, characterized in that the flow directions of the loading zone through the respective indicator zones to an absorption region (flow paths) are essentially parallel and at least two different flow paths are present. Device according to claim 1, characterized in that the indicator areas are arranged in such a way that the sample liquid does not pass through more than one indicator zone per flow path. Device according to claim 1 or 2, characterized in that the indicator areas are arranged in a diagonal row, V-shaped, W, M, N or linear. Device according to one of Claims 1 to 3, characterized in that the indicator areas comprise antibodies respectively fragments of antibodies and / or lectins respectively fragments thereof. 5. Device according to one of the claims 1 to 4, characterized in that the indicator zones comprise in particular antibodies respectively fragments of anti-A, -B, -AB, -D, -D, -C, -c, -E, -e, -Cw, and /okay. Device according to one of the claims 1 to 5, characterized in that the membrane preferably consists of polyethylene, nitrocellulose or nylon. Device according to one of claims 1 to 6, characterized in that at least one sealing element is disposed in the membrane behind the loading area and before the indicator areas. Device according to one of claims 1 to 7, characterized in that the components of the device are fixed in a carrier layer as mechanical reinforcement. 9. Device according to one of the claims 1 to 8, characterized in that the components of the device are integrated in a box. 10. Use of the device according to one of claims 1 to 9 for the blood analysis, in particular for the determination of antigens respectively epitopes of blood group antigens. Use of the device according to one of claims 1 to 10 for blood analysis, in particular for the simultaneous determination of the antigens respectively epitopes of blood group antigens A, B, AB, D, C, c, E, e , Cw and / or K. 1

2. Method for the determination of several analytes or their derivatives in a liquid sample, comprising: the application of the sample in the loading zone of a membrane of the device according to one of the preceding claims 1 to 8, this sample being present in a sufficient volume to cause the sample liquid to flow towards the absorption region through the indicator zones and to cause the analytes or their derivatives in the sample liquid to form complexes in the indicator areas. The method according to claim 12, characterized in that the analytes are antigens respectively epitopes of blood group antigens. Method according to claim 12 or 13, characterized in that the analytes comprise in particular antigens respectively epitopes of blood group antigens A, B, AB, D, C, c, E, e, Cw and / or K. 15. Method according to one of claims 12 to 14, characterized in that the antigens respectively epitopes of blood group antigens A, B, AB, D, C, c, E, e, Cw and / or K are determined simultaneously. Method according to one of claims 12 to 15, characterized in that the indicator particles are erythrocytes. Method according to one of claims 12 to 16, characterized in that the membrane is washed after applying the indicator particles. 18. Method according to claim 17, characterized in that the washing buffer is preferably hypo-osmotic. Method according to one of claims 12 to 18, characterized in that the liquid sample consists of blood or blood components, preferably whole blood, erythrocyte concentrate or a test liquid as control blood. SUMMARY The invention relates to a device for the simultaneous qualitative or quantitative determination of several analytes in a liquid sample, comprising a membrane (2) with a loading zone (5) for the application of a liquid sample, at least two indicator zones which can enter into reciprocal reaction with the analyte (s) and at least one absorption region (3) that absorbs the fluid after passing through the indicator zones, the indicator zones being located between the zone (5) ) and the absorption region (3), characterized in that the flow direction (flow paths) extend essentially parallel to the application area (5), passing through each indicator area to the region (3) Absorption At least two different flow paths are present. The invention also relates to a method for determining various analytes or derivatives thereof in a liquid sample comprising: the application of the sample in the loading area (5) of a membrane of the device according to preceding claims 1 to 8, wherein the referenced sample is present in sufficient amounts to allow the fluid sample to flow in the direction of the absorption region (3) through the indicator zone and to allow the analyte or derivatives thereof in the liquid sample to form a complex in the indicator area.