WO2004069411A1

WO2004069411A1 - Analyte detection device

Info

Publication number: WO2004069411A1
Application number: PCT/AT2004/000036
Authority: WO
Inventors: Walter Pils
Original assignee: Walter Pils
Priority date: 2003-02-05
Filing date: 2004-02-04
Publication date: 2004-08-19
Also published as: EP1592506A1; AT500380A2

Abstract

The invention relates to an analysis device (1) for determining at least one analyte from a sample, said device comprising a reagent container (2) and a container (3) equipped with an analysis section (5) consisting of a material that is capable of capillary action. The two containers have a fluidic connection in the form of a penetration device (4), such as e.g. an open end of a capillary. At least one reagent chamber (13) for the reagent and/or at least one analyte chamber for receiving the analyte container is/are located in the reagent container (2) and can communicate by means of a fluidic connection.

Description

Device for the detection of an analyte

The invention relates to an analysis device for determining at least one analyte from a sample, comprising a reagent container and a container in which an analysis section made of capillary material is arranged, and which is provided by a penetration device, e.g. an open end of a capillary that is connected to the flow, a method for determining at least one analyte from a sample with an analysis device consisting of a reagent container and a container in which an analysis section made of capillary-capable material is arranged, which are brought into contact with one another, and the use of a such analysis device.

Chemical and biochemical analyzes of liquids and solids are traditionally carried out in specialized laboratories. Nevertheless, the classic methods of analytical chemistry are now being replaced by automated analysis devices. These evaluations are typically still carried out in specialized institutions by highly trained personnel using expensive evaluation equipment. Recently, however, there has been a trend to develop devices with which it is possible to analyze samples outside of these specialized institutions and the analysis can also be carried out by less trained personnel. These rapid tests should not be manipulable by the consumer and the result should nevertheless not be available in coded form.

The analysis of body fluids, e.g. Blood or urine is a simple undertaking unless only a small amount of fluid can be obtained, as is the case with saliva, for example. The small amounts of saliva that are secreted into the mouth also have a high viscosity, in contrast to blood or urine.

Nitrocellulose strips are available for the analysis of body fluids, such as urine. On the one hand, these devices are inexpensive and, on the other hand, they produce rapid yet reproducible results for various applications. Due to the high viscosity of These strips of saliva cannot be used to analyze them. It is known that saliva, which is analyzed directly and undiluted using a nitrocellulose strip, has poor migration properties on these strips due to the relatively high concentration of mucin and other viscous protein-containing substances.

The analysis of saliva has not been intensively investigated in recent years because blood and urine samples were available as primary liquids for the detection of different analytes or for the testing of various diseases. However, it is now known that the concentration of lymphocytes, plasma cells and immunoglobulins in saliva is directly related to that in the blood. In addition, there are immunoglobulins in the saliva that are specific to the saliva, e.g. IgA. In contrast to blood, urine and saliva can also be obtained from medically untrained personnel. There are various techniques for collecting saliva, such as with capillary tubes, micropipettes, by chewing paraffin or by spitting into a tube.

US 6,008,056 A describes a method and an apparatus for examining a certain volume of a sample on a chromatographic strip. The device comprises a sample reservoir for receiving the sample, an overflow device for the excess sample liquid, a chromatographic strip and a recess in communication with the overflow device. The apparatus has a piston for compressing a sample pad which is pressed against the pad. By pressing against the sample pad, the sample accumulated in the pad is drained through an opening. Typically, the sample pad compressor is a modified syringe. The sample is brought together via an outlet device in a chamber, which can contain two solutions. The sample now reaches the sample reservoir from the chamber via a connection. The sample reservoir is surrounded by a recess that collects the excess liquid. A disadvantage of this device is that a certain sample volume has to be collected in order to be able to squeeze out the sample collection pad. If solid starting materials or small sample volumes are present, there is no possibility of carrying out an analysis using this device.

No. 5,935,864 A describes a method and an analysis kit for collecting liquid samples consisting of a sample container and a reagent tube. The sample container has an open and a capillary end with a chamber in between. The reagent The tube is sealed with a penetrable film and is used to hold the capillary end of the sample container. In order to carry out the analysis, the capillary-shaped end, which had previously been brought into contact with the sample, penetrates the film of the reagent tube. The liquid from the capillary can be drawn into the sample container. The analysis kit can also consist of a further chamber, which contains a buffer solution. The buffer chamber is also closed with a septum that can be pressed in. The sample container comprises a stamp which fits into the reagent tube and thus exerts pressure on the buffer solution contained therein and the contents of which are sucked through the capillary. The liquid buffer mixture now enters the sample container, where the sample can be analyzed. Transferring the buffer solution to the sample is made possible by positively bringing the sample container together with the buffer chamber or the reagent tube. Only liquid samples can be analyzed with this device.

A disadvantage of US 5,935,864 A proves that the sample, e.g. Blood enters the capillary directly from the patient and is subsequently transferred directly to the test strip via the reagent. However, the high concentrations of organic material in the blood can lead to disturbances, especially interference, during the analysis. In addition, the sample is exposed until immediately before the analysis is carried out, and there is therefore an increased risk of contamination for the personnel performing the analysis.

A disadvantage of these devices known from the prior art is that only one analysis test strip is available in each case and thus neither an increase in the sample throughput is possible nor a large number of analytes can be detected at the same time.

The object of the invention is therefore to provide an analysis device with which the detection and determination of analytes can be carried out quickly, easily and reproducibly. Part of the object of the invention is also to minimize the risk of contamination before and during the analysis.

The object of the invention is achieved independently in each case by an analysis device mentioned at the outset, in which at least one reagent chamber for the reagent and at least one analyte chamber for receiving the analyte are arranged in a flow-connectable manner in the reagent container, or by a method mentioned at the outset, which enables the reagent container and the like Container, which are already arranged next to each other (arranged next to each other) by means of a pen Fration device to create a flow connection, wherein the reagent container and the container are moved relative to each other, such as shifted, rotated or folded, or by the aforementioned use of said analysis device for detecting at least one analyte from a group comprising active ingredients, hormones, proteins, peptides , Allergens, antigens, antibodies, neurotransmitters, nucleic acids, carbohydrates and / or lipids. The advantage here is that the reagent and analyte pass from the reagent container into the container and the reagent and analyte are mixed or mixed and the reagent-analyte mixture is thus already present at the beginning of the analysis section. The penetration device establishes a flow connection between the container and the reagent container as soon as the positions of the reagent container and the container relative to one another are changed, such as reduced, for example, and the analyte is introduced into the analyte chamber and thus the reagent-analyte mixture can flow to the analysis section. The short-term combination of the reagent with the analyte immediately before the addition to the analysis section prevents the analyte in the reagent from being changed or degraded and thus no longer being available in its original state at the start of the analysis. All reagents required for analysis are in the reagent container. It proves to be advantageous that the reagents can be present both in separate reagent chambers and in one reagent chamber. In the case of separately available reagents, no undesired reactions of the reagents take place before the start of the analysis, but the reactions only take place in the presence of the analyte. When the reagents are present in a reagent chamber, it proves advantageous that only one reagent chamber has to be penetrated and therefore the reagents from many different reagent chambers do not have to be brought into contact with the analytes before the reagent-analyte mixture is transported to the analysis section, but rather the one Analyte only needs to be brought into contact with the reagent from a reagent chamber.

It also proves advantageous that a large number of different analytes can be detected with the same analysis device, as a result of which a simple means for analyzing samples is available through the use of the analysis device.

Furthermore, it proves to be of advantage that the use of the analysis device enables a quick and uncomplicated andyte determination, regardless of the location, and that a closed system is available for the analysis and thus the risk of contamination is reduced. According to an embodiment variant, it is provided that a means for connecting the reagent container to the container, such as a tongue and groove connection, a thread, a bayonet connection and / or predetermined breaking point, etc., is arranged, as a result of which there is firm cohesion. It has been found to be advantageous that the analysis device is ready for use, thereby avoiding errors that can occur when assembling or assembling variable analysis devices. By arranging the connecting means, the containers can be separated for subsequent evaluations if necessary.

The one or more penetration devices of the analysis device can be designed as a cannula, tip, wick, spike, etc., after which a flow connection can be established between the reagent container, the reagent chamber, the analyte chamber and the container, which can be used to transfer the reagent from the reagent chamber the analyte chamber serves in the container. Due to the time-delayed establishment of the flow connection, the analyte-reagent mixture does not come into contact with the analysis section until immediately before the analysis and therefore no unspecific reactions can take place in advance. Furthermore, it proves advantageous that the analyte-reagent mixture can already be mixed in the penetration device. By arranging several penetration devices, it can be achieved that the analyte-reagent mixture reaches the analysis section directly and thus the transport via another capillary material can be avoided and the sample loss is minimized.

It also proves to be particularly advantageous that the same device can be used for the penetration of the reagent container, the reagent chamber and the analyte chamber and for the transfer of the reagent into the container.

In a further development of the embodiment variant of the invention just mentioned, it is provided that a capillary-shaped object does not necessarily have to be used as the penetration device, but the penetration can also take place, for example, through an object which has capillary-capable properties.

It is also possible that the wall of the reagent container, the reagent chamber, the analyte chamber and / or the container is at least partially formed by a septum, the analysis of the analyte being able to take place with a time delay after the sample has been taken or the sample has been fed into the analyte chamber and therefore several samples taken at different times collected, can be analyzed together under the same framework.

A predetermined breaking point for the transfer of the reagent into the analytical chamber and / or the container can be formed in the wall of the reagent container, the reagent chamber, the analyte chamber and / or the container, after which the opening takes place at a predefined stand and does not take place arbitrarily and thus that Unreacted reagent passes from the reagent container, the reagent chamber or the analyte chamber into the container. The targeted transfer of the reagent at a predefined location enables the reagent to be passed on to the container by means of the penettation device.

At least one positionable closure element can be arranged on the reagent container, wherein the reagent remains enclosed in the reagent container in the reagent chamber regardless of the position of the analysis device, thus reducing the risk of contamination for the environment and the personnel performing the analysis.

In a further development of the embodiment variant of the invention just mentioned, it is provided that at least two closure elements can be arranged in parallel, according to which a tight analysis device is still present even when the reagent container moves relative to the container, and contamination-free working can thereby still be ensured.

According to an embodiment variant, it is provided that a flap which can be pivoted through 90 ° can be arranged in the analyte chamber, the horizontal arrangement of the flap on the one hand preventing premature unintentional changes in the position of the two containers relative to one another and on the other hand preventing contamination of the analyte chamber before analysis becomes. Due to the vertical arrangement of the flap after the analyte is fed into the analyte chamber, the analysis device can be kept closed and the analyte can no longer emerge from the analysis device. This also proves to be advantageous that, regardless of the position of the analysis device, there is no risk to the environment, the subject or the analyzing personnel.

The reagent container can be arranged to be movable on the container, such as displaceable, rotatable and / or foldable, after which a penetration of the reagent container only takes place after the application of a force takes place and the reagent-analyte mixture is transferred into the container and can thus reach the analysis section.

According to a variant of this, provision is made for a position control in the reagent container and / or container to check the relative movement, such as e.g. the sliding, rotating and / or folding movement, the container and the reagent container is formed, after which any manipulation of the reagent container in relation to the container can be controlled. This enables the transfer of the reagent from the reagent container into the container to be controlled and a reagent is therefore always available for the analysis section.

According to a development of the analysis device according to the invention, several analysis sections for different analytes can also be arranged in the container, according to which a large number of analytes can be detected simultaneously and the analysis device cannot be used exclusively for the detection of a specific analyte. As a result, the production costs for the different configurations of the analysis devices can be minimized. It also proves advantageous that the same analysis device can be used for the detection of all possible analytes.

In particular, the formation of the analysis section from capillary material has proven to be particularly advantageous, the analyte-reagent mixture being transported by the physical properties of the analysis section and no additional means for transporting the analyte-reagent mixture from the reagent container into the container having to be arranged.

The analysis section can consist of at least one analysis field, whereby simple analysis results, e.g. only a color reaction, such as pH value determinations, can be read.

A strip of absorbent material can be arranged in the container at at least one end of the analysis section, as a result of which the reagent is transported from the reagent container through the capillary action of the absorbent material into the container and is thus fed to the analysis section. It proves to be advantageous that no mechanically or mechanically operated devices for transferring the reagent from the reagent container into the container are required, and thereby a cost-efficient execution of the analysis device is made possible. In a further development of the embodiment variant of the invention just mentioned, it is provided that the strip is arranged at two diametrically lying ends of the analysis section. This improves the conductivity of the reagent from one end of the analysis section to the other end and thus the separation of the analyte. Another advantage is that a targeted transport of the reagent-analyte mixture is made possible.

In a further development of the analysis device, the strip is designed as a penetration device, the arrangement of an additional device for penetrating the reagent container, the reagent chamber and the analyte chamber being saved. It also proves to be advantageous that the capillary action already transfers the reagent from the reagent container into the container and the reagent-analyte mixture is thus immediately available at one end of the analysis section.

The formation of a recess in the analysis device above the strip makes it possible for the analyte to be determined to be applied directly to the strip and only have to be washed into the analysis section via the reagent, and because of the short transport distance there is no or negligible sample loss comes.

The analysis section can be divided into a start zone and at least one run, dye, conjugate, control and / or analysis field, with the separation of the analyte in the different zones reacting with different reagents which are immobilized in the different fields , can take place and these can then be detected in the analysis field. Another advantage is that, regardless of the presence of an analyte in the sample, a mark is detected in the zone of the control field in order to control the functionality of the analysis device. A further advantage is the temporary immobilization of the reagents in the various fields, in particular in the dye field and in the conjugate field, because this eliminates the need to add these reagents during the analysis, thus simplifying the process and preventing confusion of the reagents. The arrangement of running fields advantageously ensures that the analytes are freed from particulate contaminants during the passage of the capillary-capable material.

In the analysis field, at least one analyte and / or an analyte-specific binding partner can be immobilized in a predefined concentration, wherein at least a semi-quantitative determination of the analyte to be detected can be made possible by using several analysis fields which are arranged either on an analysis line or in adjacent analysis lines, in each case a different concentration of the analyte or analyte-specific binding partner is immobilized. The arrangement of different concentrations on adjacent analysis sections also proves to be advantageous that there is an additional control option for the analysis method, because if the detection of a low concentration fails, an error in the analysis can be detected, whereas in the case of a linear arrangement of several analysis fields an analysis section cannot be checked whether there is possibly too little reagent for the transport of the analyte from the start zone into the analysis section or whether there is actually a lower concentration of the analyte.

It is also possible that at least parts of the analysis field of the analysis section are arranged visibly in the container, wherein the analysis of the analysis can take place directly in the analysis device and the analysis section does not have to be removed from the analysis device. This offers the advantage that the personnel carrying out the analysis and the surroundings cannot be contaminated by the analysis reagents or by the analyte.

One or more markings can be arranged on the container to support the evaluation of the analysis, according to which, when evaluating the analysis, it is only necessary to check whether there is a marking on the analysis line at the position to be expected, which is identified by a marking on the analysis device occurs. As a result, a quick statement about the result can be made by simply comparing the markings on the analysis device and the analysis section.

According to a variant, a fixing device, such as e.g. Arrange a snap-in device, which attaches the reagent container to the container in a predefined position, on the container and / or on the reagent container, with which an unintentional adjustment of the container relative to the reagent container can be avoided during the analysis and thereby a controlled transfer of the reagent from the reagent container into the container can be guaranteed.

One or more auxiliary substances for carrying out the analysis, such as stabilizers, preservatives and / or drying agents, can be present in the container because this can extend the shelf life and the usability of the analysis device. One or more analytes from a group comprising active substances, hormones, proteins, peptides, allergens, antigens, antibodies, neurotransmitters, nucleic acids, carbohydrates and / or lipids can be arranged in the analyte chamber, after which many different analytes can be detected. It proves to be advantageous that a variety of analytes can be detected with the same analysis device without having to adapt the analysis device. Another advantage is that, due to the diversity of the analysis device, no special training of the personnel performing the different analytes is required.

Active substances from a group comprising drugs, drug substitutes, performance-enhancing substances such as doping agents, pharmaceuticals, toxins or their metabolites can be arranged in the andyt chamber, after which, in particular, analytes which influence the physiological balance of humans can be detected. It has proven to be advantageous that the analysis device ensures simple handling and can therefore be carried out by anyone. This multiply applicable analysis device minimizes costs for the analysis of the most diverse analytes. The analytes can be determined directly on site and do not have to be transported to a special laboratory. The rapid test result also enables an early decision to be made as to whether the analyte is impairing the individual and thus further dangerous situations can be avoided. A rapid test result also prevents the possibility of manipulating biological samples, for example by taking doping maskers. The immediate availability of a result on site makes it much more difficult for the test person to provide evidence.

Both drugs and psychotropic drugs impair the respondent's ability to react at the workplace and their ability to drive in traffic. The immediate danger of such situations can be recognized immediately by obtaining a rapid test result using this method.

The invention is explained in more detail below with reference to the exemplary embodiments shown in the drawings.

Show it:

1 is a top view of an analysis device 1; 2 shows an embodiment variant of an analysis device 1 according to the invention;

3 shows a further embodiment of an analysis device 1 according to the invention;

4 shows a variant of the inventive analysis device 1 according to FIG. 3.

It should be noted that in the differently described embodiments, the same parts are provided with the same reference numerals or the same component names, and the disclosures contained in the entire description can be applied analogously to the same parts with the same reference numerals or the same component names. The location information selected in the description, e.g. above, below, laterally, etc. refer to the figure described and illustrated immediately and are to be transferred analogously to the new position in the event of a change in position. Furthermore, individual features or combinations of features from the different exemplary embodiments shown and described can also represent independent, inventive or inventive solutions.

1 shows a schematic illustration of an analysis device 1. The analysis device 1 consists of a reagent container 2 and a container 3, the reagent container 2 and the container 3 being arranged next to one another. To establish the flow connection between the two containers a Penefrationseinnchtung 4 is arranged.

1, the penetration device 4 is formed by a cannula, l alternative embodiments, the penetration device 4 can be formed by any pointed or angular object, such as Tip, spike, wick, strips, etc. When forming the penetration device 4 as a strip made of a capillary material, such as e.g. Nitrocellulose, foam, etc., or as a solid hollow object, which can be pointed, filled with a capillary material, the reagent or reagent-analyte mixture is transferred from the reagent chamber 13 into the container 3 to the analysis section 5 via its capillary-active properties reconciled. When the Penefrationseinrichtui g 4 is designed as a wick or spike, a channel, which represents a connection between the reagent chamber 13 and the starting zone 6 of the container 3, can be formed in the reagent container 2. The penetration device 4 is arranged either on the reagent container 2 or on the container 3.

The shape and the outer dimensions of the analysis device 1 can vary depending on the use. The outer dimensions of the container 3 are preferably insignificantly smaller than that of the reagent container 2 in order to at least partially shift the two containers 2, 3 into one another to enable. The outer shape can be angular, as shown in FIG. 1, or it can also take on round or ellipsoidal shapes, as in a variant not shown.

The connection or the merging of the reagent container 2 and the container 3 takes place either via webs at which a predetermined breaking point is provided in order to be able to carry out a relative movement of the two containers 2, 3, or it is a tongue and groove connection, a thread, a bayonet connection, em hinged hinge or the like.

The analysis device 1 is preferably made of plastic, such as e.g. Polystyrene, cycloolefin copolymers (COC), polypropylene, acrylic butadiene styrene, polyamide, polycarbonate, polymethyl methacrylate, polysulfone and or styrene acrylonitrile, etc. are manufactured. A combination of two or more materials or different plastics can also be used, for example 2 COC being used for the reagent container and 3 polystyrene for the formation of the container. In an alternative embodiment, the reagent container 2 and the container 3 are formed from COC and the reagent chamber 13 from polystyrene.

In the container 3 is the analysis section 5, which is formed from an absorbent or capillary material. The capillary-capable material, which represents a filter, consists of cellulose and / or its derivatives from organic polymers and their derivatives, in particular from materials that adsorb little protein on their surface, such as e.g. Glass fiber, ceramics, polytefrafluoroethylene (PTFE), nylon, polyvinyl fluoride (PVDF), silicon-based materials, materials of biological origin, acetates or from mixtures or modifications of these materials.

The filters are selected with a pore size selected from a range with an upper limit of 50 μm, preferably 40 μm, in particular 35 μm, and a lower limit of 0.2 μm, preferably 5 μm, in particular 10 μm. Filters with a pore size selected from a range with an upper limit of 30 μm, preferably 25 μm, in particular 20 μm, and a lower limit of 12 μm, preferably 15 μm, in particular 17 μm, have also proven to be particularly advantageous.

Pre-filters with an undefined pore size can also be used in particular in the area of the start and target zones 6, 7 and in the area of the running areas 8. The capillary-capable material is formed by a filter with an arbitrarily defined pore size. In an alternative embodiment, a recess, which serves to receive the analyte, can be formed in the analysis device 1 in the region of the start zone 6. The recess can be designed both for receiving the analyte in liquid and in solid form. The start zone 6 can be constructed in the area of the recess from several layers of filters in order to remove or minimize impurities from the sample before the analysis.

The capillary-capable material of the analysis device 1 has a thickness selected from an area with a lower limit of 0.05 mm, preferably 0.08 mm, in particular 0.15 mm and an upper limit of 2 mm, preferably 1.5 mm. in particular 1 mm. A thickness selected from a range with a lower limit of 0.1 mm, preferably 0.15 mm, in particular 0.2 mm and an upper limit of 0.9 mm, preferably 0.8 mm, in particular 0, has proven particularly advantageous , 5 mm. The thickness of the capillary-capable material can vary in the different fields, i.e. that the analytical device 5 does not have to have a uniform thickness over its entire extent. The various analysis fields 10 can consist of the same absorbent material or of different materials. The thickness and pore size of the absorbent material can also be designed differently in the various fields.

At one end of the analysis section 5, one or more further strips of absorbent material, which are arranged normal to the analysis section 5, are formed. The start and destination zones 6, 7 can be arranged on these strips. In addition to the materials already mentioned, these strips can also consist of a pre-filter with an undefined pore size. The strips can also be formed from different capillary materials or from the same material but with different pore sizes in order to achieve different running properties, e.g. a different speed of achieving the sample on the individual strips.

The analysis track 5 and the start and finish zone 6, 7 in the container 3 are connected via a connection layer, such as e.g. a double-sided adhesive film, connected to the container 3. The connection of the analysis section 5 and the start and target zones 6, 7 to the container 3 can also be made using a one-sided adhesive carrier material (laminating).

Several fields can be arranged on the analysis section 5, such as a conjugate field 9 with specific binding partners to which at least one substance is conjugated, at least one field 8, an analysis and control field 10, 11 on which different analytes can be bound and / or a dye field 12.

The different fields can be in different forms, e.g. linear, circular, triangular, quadrangular, hexagonal, pentagonal, heptagonal, octogonal, frapezoid, rhomboid, etc., can be arranged on the analysis section 5.

The number and order of the fields can also vary. For example, only one analysis field 10 or only a combination of an analysis field and a control field 10, 11 can be arranged on the analysis section 5, or the conjugate 9, dye 12, analysis 10 and control field 11 are directly strung together and are thus there are no running fields 8, or several analysis fields can be arranged for a different analyte.

The analysis device 1 has a recess at least above the analysis field 10 and or the control field 11 or is formed at this point by a transparent material so that the result of the analysis can take place without disassembling the analysis device 1.

For better evaluation of the analysis device 1, several markings can be attached to its surface, for example, which predefine the position of the expected result and thus enable faster orientation on the analysis section 5.

To saturate the free non-specific binding sites, the capillary-capable material is treated with solutions containing proteins such as BSA, milk powder, casein, gelatin, fat-free milk powder, calf serum, etc. and / or with commercial blocking reagents, such as e.g. Blotto or Superblock (from Pierce) in a concentration of 0.1 mg / ml to 10 mg / ml and / or detergents, such as e.g. Tween, Triton, Nonidet P-40, Chaps, etc., in a concentration of 0.005% by weight to 5% by weight, blocked.

In the analysis field 10, at least one analyte or an analyte-specific binding partner is selected in a concentration from a range with a lower limit of 0.5 ng / ml, in particular 1 ng / ml, preferably 2 ng / ml, and an upper limit of 5 mg / ml, in particular 3 mg / ml, preferably 1 mg / ml, immobilized. The at least one analyte can be detected both directly and indirectly, such as by the analyte itself, a specific analyte binding partner or a labeled binding partner. Indirect detection is carried out by binding the analyte to a first specific binding partner, eg primary antibody. In analysis field 10, either this or the analyte (on another stand) is then recognized and bound by a second specific binding partner, for example secondary antibody, which is immobilized in analysis field 10. The presence of the analyte in the analysis field 10 is finally linked to a further specific binding partner which is conjugated to a substance which either recognizes the analyte itself (at another location) or the analyte bound to the first specific binding partner or the second specific binding partner. demonstrated.

At least one analyte is immobilized in the analysis field 10. As an alternative to the analyte, an analyte-specific binding partner can also be immobilized in the analysis field 10. In this case, for the detection of the analyte, the specific binding partner which is conjugated to a substance must be specific for the analyte.

The analyte can be introduced into the axial chamber 14 either in the liquid state or in the solid state. A variety of different analytes can be detected. The at least one analyte can be selected from a group comprising drugs or drug substitutes including cannabis products such as marijuana, hashish and / or cannabinol, cocaine such as benzoylecgonine, crack and / or crystal, opiates such as morplium, acetylmoφhin, heroin, codeine, Propoxyphene and / or fentanyl, nicotine, kotinin, d-lysergic acid diethylamide (LSD), psilocybin and psilocin, mescaline and peyote, methadone or methadone metabolites, or a designer drug such as amphetamines (MDA, dimethoxvbromamphetamine, etc.), me e.g. ecstasy, MDMA), phencychdin (angel dust), y-hydroxybutyric acid (liquid ecstasy), anti-epileptics, such as phenytoin, doping agents from a group comprising anabolics, such as testosterone and its derivatives, somatofropin, ephedrine derivatives, analeptics, such as strychnine , Amphetarnine derivatives, analgesics, antitussives, agents for increasing the oxygen transport capacity and the availability of oxygen for the skeletal muscles, such as erythropo ietin and / or diuretics, pharmaceuticals, analgesics or psychotropic drugs from a group comprising neuroleptics, such as phenothiazine, butyrophenones and / or thioxanthenes, antidepressants, such as MAO (monoamine oxidase) inhibitors, inhpramine, desipramine, amitriptyline, etc., tranquilizers, such as z. B. benzodiazepines (diazepam), barbiturates and / or psychoanaleptics, stimulants such as phenylethylamine, antiepileptics and / or hypnotics, antibodies, formed in response to viral, viroid, bacterial, mycotic, parasitic or prion-based infections and / or formed as a result of immunizations (vaccination in humans or animals or polyclonal antibody production in animals) and / or formed as a result of autoimmune diseases tack or allergic reactions, hormones such as HCG (human chorionic gonadotropin), proteins as tumor markers from a group comprising squamous cell carcinoma antigen (SCC), thyroglobin (Tg), steroid hormone receptors, prostate-specific antigen (PSA), neuron-specific enolase (NSE), Carcinoembryonic Antigen (CEA), Alpha-Fetoprotein (AFP), CYFRA 21-1, CA 125, 19-9, 72-4, 15-3 and / or human Calcitonin (hCT), MCA (Mucine-like cancer associated antigen), toxins from a group comprising botulinum toxin, toxins from animals, plants, such as: fungi, bacteria, algae, plants and their constituents, foods and / or toxins of artificial origin, for example combat poisons, and / or heavy metals, herbicides, fungicides , Pesticides, bactericides, antibiotics, preservatives, etc. can be selected. Furthermore, allergens such as. e.g. pollen, dust mite droppings, histamine, etc. can be detected. The detection of prions from the nervous system of various mammals as well as from feed or food is of increasing importance to prevent the spread of Creutzfeldt-Jakob disease. Markers or species-specific proteins which are used in genetic engineering to identify genetically modified foods, seeds, etc. can also be detected as analytes. Various types of antibodies (sub) can also be detected.

The analyte can be immobilized in several analysis fields 10 in different concentrations. Due to the concentration of the analysis fields 10, a semi-quantitative determination of the analyte can be carried out. The concentration of the at least one analyte in the dilution series decreases with a factor selected from a range with a lower limit of 1, in particular 2, preferably 3 and an upper limit of 100, in particular 10, preferably with a factor 5.

In an alternative embodiment, a different concentration of the analyte for the semi-quantitative determination of the analyte can be immobilized in several analysis areas 5. For example, in successive analysis areas 5, which are arranged in the analysis device 1, the concentration of the analyte can be multiplied.

The analyte can be obtained from a sample of biological origin, such as body fluids from humans and animals, such as blood, plasma, serum, urine, saliva, sweat, sperm, etc. and / or vegetable origin, such as leaf, fruit, seeds, etc. and / or microbiological origin. The analyte can also be detected from the soil or from water. In order to carry out an analysis of substances from the air, the analyte has to be concentrated, for example by sucking in larger air volumes by means of a pump, and the analyte can be enriched directly on the material which is fed to the analyte chamber 14.

n Dye field 12, one or more dye components or their precursors are temporarily immobilized. However, the dye components do not necessarily have to be in one zone, but can also be divided into several zones, e.g. on the starting zone 6, on the dye field 12, on the conjugate field 9 and the running field 8. The dye components are in undissolved form. The dye components used and their precursors are: tyramine and its derivatives, X-gal (5-bromo-4-chloro-3-indolyl-ß-D-galactoside), S-gal (3,4 cyclohexenoescetin-ß-D -Galactopyranoside), ONPG (o-nitrophenyl-ß-D-galactopyranoside), CPRG (chlorophenol red-ß-D-galactopyranoside), bluogal (5-bromo-3-hidolyl-ß-D-galactoside), MUGal (4- Methylumbelhfeιyl-ß-D-galactopyranoside), NBT (nitroblue tetrazolium chloride), BCJP (5-bromo-4-chloro-3-h dolylphosphate), PNPP (p-nitrophenyl phosphate), OPD (o-phenylenediamine), ABTS (2nd , 2'-azino-di- (3-ethylbenzothiazoline sulfonic acid), DAB (3,3-diaminobenzidine), TMB (3,3 '5,5'-tetramethylbenzidine), phenazutethosulfate, potassium ferrocyanide, potassium ferricyanide, ferric ammonium cifrate, VectorBlackTM, chemiluminescent dyes, such as Galacton-Star substrate, Lumi-PhosTM plus, LuciGLOTM, DuoLuXTM, Lumigen PS-3, Chemiluminescent HRP (horseradish peroxidase) substrate, 2 component system from BioFXTM, and / or Lumi-Gal 530 and / or Fluoureszenfarbstoffe, e.g. 3-carboxy umbelhferyl-β-D-galactopyranoside (CUG) and / or MUP (4-methyl umbelhferyl phosphate). The concentration of the dye components and their precursors is, in each case in% by weight, from a range with a lower limit of 0.01%, in particular 0.1%, preferably 1%, and an upper limit of 20%, in particular 10%. preferably 4% selected.

In the field with the dye components, other substances, such as, for example, BSA, maltrine, milk powder, casein, gelatin, can also be selected with a concentration from a range with a lower limit of 0.1%, in particular 1%, preferably 5%, and an upper limit Limit of 50%, in particular 30%, preferably 10%, are added in order to reduce the adsorption of the dye components to the analysis device 1 or to increase the solubility of the dyes. In an alternative embodiment, the dye field 12 is absent, the reagents for dye reaction being contained in the reagent of the reagent chamber 13.

In conjugate field 9 there are in particular specific binding partners, e.g. Antibodies to analytes, which are supported with a catalyst, e.g. an organic or inorganic catalyst or enzymes or proteins, e.g. β-galactosidase, peroxidase, alkaline phosphatase, sfreptavidin / avidin, protein A or similar molecules capable of high-affinity specific bonds, etc., are conjugated. Alternatively, several different substances can also be conjugated to a specific binding partner simultaneously or in a predefined sequence. The concentration of the specific binding partners to which the substance is conjugated is selected from a range with a lower limit of 0.5 ng / ml, in particular 1 ng / ml, preferably 2 ng / ml, and an upper limit of 5 mg / ml, in particular 2 mg / ml, preferably 1 mg / ml. Concentrations have also been selected particularly advantageously from a range with a lower limit of 0.1 mg / ml, preferably 0.2 mg / ml, in particular 0.3 mg / ml, and an upper limit of 2 mg / ml, preferably 1 mg / ml, especially 0.5 mg / ml.

Control field 11 contains an immobilized specific binding partner, e.g. an antibody for at least one substance, e.g. β-galactosidase, alkaline phosphatase, peroxidase, streptavidin / avidin, protein A or similar molecules capable of highly specific bonds, etc., in a concentration selected from a range with a lower limit of 0.5 ng / ml, in particular 1 ng / ml, preferably 2 ng / ml, and an upper limit of 5 mg / ml, in particular 3 mg / ml, preferably 1 mg / ml.

It is not necessary to immobilize a substance or an analyte in the start and target zones 6, 7 and the running fields 8.

Several analytes or combinations of, e.g. B. specific binding partners, substances and / or dye components or their precursors.

For a better overview, the individual fields of the analysis section 5 are not shown in FIGS. 2, 3 and 4. However, it goes without saying that these can be arranged as shown in FIG. 1 or described in FIG. 1.

FIG. 2 shows a top view of the analysis device 1, wherein a reagent chamber 13 for receiving the reagent is formed in the reagent container 2. The reagent can be stored in the nis 2 or in a separately designed reagent chamber 13. The reagent chamber 13 is preferably arranged at the opposite end of the reagent container 2 with respect to the penetration device 4. The reagent chamber 13 can be fastened in the reagent container 2 by means of sealing materials, such as, for example, foam-like, rubber-like or other flexible materials. The reagent container 2 can also be sealed with sealing material for better sealing.

In an alternative embodiment variant, not shown, a plurality of reagent chambers 13 can also be arranged in the reagent container 2, wherein different reagents can be located in the respective reagent chambers 13.

The total volume of the reagent is selected from a range with an upper limit of 10 ml, preferably 8 ml, in particular 6 ml and a lower limit of 100 μl, preferably 500 μl, in particular 1 ml. A range with an upper limit of 5 ml, preferably 4 ml, in particular 3 vA, and a lower limit of 1.5 ml, preferably 2 ml, in particular 2.5 ml, has also proven to be particularly advantageous.

hn container 3, an absorbent material is arranged on the two diametrically arranged sides of the analysis section 5. The start zone 6 made of capillary-capable material serves to transfer the reagent from the reagent chamber 13 in the reagent container 2 via the penetration device 4 to the analysis section 5. The capillary-capable material in the target zone 7 draws the reagent-analyte mixture from the start zone 6 over the analysis section 5 ,

3 and 4 show a further embodiment of the analysis device 1, an analyte chamber 14 being arranged in the reagent container 2. The analyte chamber 14 serves, directly or indirectly, via the reagent-analyte mixture, to hold the analyte which is to be analyzed in the further analysis.

The analyte, which is directly or bound to a carrier 15, such as cotton swabs or the like, is fed through a closure element 16 into the analyte chamber 14, which is arranged on the reagent container 2. The closure element 16 is arranged in FIG. 3 on the side wall of the reagent container 2, but it can of course also be arranged on the top or bottom wall. The closure element 16 can either be made in one or more parts, such as, for example, two parts. The two-part embodiment of the closure element 16 consists of two parts arranged in parallel, which are movable. In the analyte chamber 14 of the reagent container 2 there is also a flap which can be pivoted through 90 °

17 arranged, which is folded when the analyte is fed into the analyte chamber 14. The pivotable flap 17 prevents the two containers 2, 3 from moving unintentionally to one another without an analyte being fed to the analyte chamber 14, and thus fulfills a safety function, such as a lock.

In the container 3, in addition to the one analysis section 5, further analysis sections 5 can be arranged, these being parallel to the first analysis section 5. In an alternative embodiment, the analysis sections 5 can also be arranged in a circular or polygonal manner.

The reagent container 2 and the container 3 are constructed such that the arrangement of the two containers 2, 3 relative to one another is changed by the action of a force such that the penetration device 4, which is located in or between the reagent container 2 and the container 3, is in Direction of the reagent chamber 13 is displaced and penetrates it, thereby establishing a connection between the reagent container 2 and the analyte chamber 14 and the starting zone 6 in the container 3.

A position control 18 for checking the movement of the reagent container 2 relative to the container 3 can be arranged on the outside of the containers 2, 3. This position control

18 can be a marking which results in a predefined pattern, for example a recess into which an elevation is fitted, or two linear markings which produce a continuous line, or a colored marking, etc.

A plurality of fixing devices 19 can be designed to position the container 3 on the reagent container 2, e.g. Locking devices. These fixing devices 19 have the effect that it is no longer possible to subsequently move the two containers 2, 3 during the analysis, once they have been pushed into one another, pressed, folded or rotated.

To receive a controlled sample volume, an element 20 with a predefined capacity for receiving the sample from the carrier 15 can be arranged in the analyte chamber 14. This makes it possible for a certain sample volume to be available in the analysis process and thus an analysis with the same sample volume to be easily reproduced.

An embodiment variant is not shown, in which the analyte is fed directly into the reagent chamber 13 of the reagent container 2. The reagent-analyte mixture also passes through here a movement of the two containers 2, 3 relative to one another via the penetration device 4 to the starting zone 6 in the container 3, the sample to be analyzed being fed to the analysis section 5.

On or in the analysis device 1, a fastening for the arrangement of a means for sample collection can be arranged, which can be removed from the analysis device 1 by simple manipulation and with which the sample can be obtained, drawn, removed, etc.

FIG. 4 shows an embodiment variant of the analysis device 1 according to the invention, the reagent container 2 and the container 3 being displaced relative to one another. The flap 17 is pivoted by the supply of the analyte, which is bound on the carrier 15, and the container 3 can be pushed into the reagent container 2, pressed, folded or rotated. The position control 18 now shows a marking and thus confirms the execution of the movement of the two containers 2, 3. The element 20 can be compressed during the movement of the two containers 2, 3 and the sample volume can thus be transported to the analysis section with the reagent.

5 shows the arrangement of a plurality of penetration devices 4 between the reagent container 2 and the analysis sections 5 of the further container 3.

The reagent container 2 and the container 3 can also be separated from one another after the analysis has been completed, and each can be stored as a separate device, for example to enable the analysis result to be read at a later point in time.

To carry out the method, a small amount of a sample is collected with an absorbent material such as a cotton swab. The analyte adhering to the absorbent material is introduced into the analyte chamber 14. By the action of a force, the container 3 is pressed against the reagent container 2, folded or rotated, and the reagent chamber 13 is moved with the penetration device 4, such as a needle or a tip, or with a tire, on which the starting zone 6 is located. open. The reagent, in particular organic reagent, is sucked from the reagent chamber 13 via the sample into the start zone 6 of the container 3 via a channel or the absorbent material. As a result of the capillary action, the reagent-analyte mixture is drawn into the target zone 7 via the analysis line 5 and separated on the analysis line 5. The reagent with the analyte migrates in the capillary-capable material of the analysis direction 1 at a speed of 1 to 12 cm / minute from the starting zone 6, for example through the conjugate field 9, dye field 12, running field 8, analysis and control field 10, 11 and finally into the target zone 7. If sufficient in the reagent with the sample If the amount of analyte 13 is present and all the binding sites of the specific binding partners are saturated, they can no longer bind to the immobilized analytes in the analysis field 10. A detectable amount of the analyte on the analysis device 1 only gives a mark in the control field 11 because the binding partners are already saturated and do not react with the immobilized analytes on the analysis device 1, in the event that no analyte or an insufficient amount is available in the sample Detection of the analyte is present, there are two markings on the analysis device 1, the first marking resulting from the binding of the analyte-specific and / or substance-conjugated analyte-specific binding partner in the analysis field 10 and the second marking resulting from the binding of the immobilized binding partner specifically for the substance , such as ß-galactosidase, in reaction with a dye component or its precursor.

If the analyte to be determined is missing in the sample, the analyte-specific binding partner binds in the analysis field 10, and the analyte-specific binding partner to which at least one substance, e.g. β-galactosidase, conjugated, reacts with the passing dye component or its precursors in such a way that the dye or dye complex formed remains either insoluble or poorly soluble in the analysis field 10 and / or can be detected by antibodies to the dye complex formed. The specific binding partner, to which at least one substance is conjugated, can carry several binding sites for dyes or dye complex molecules that produce a clearly visible label.

If an analyte is present in sufficient quantity, it binds to analyte-specific binding partners that are conjugated with a substance, and these binding partners can therefore no longer bind to the immobilized analytes in the analysis field 10 and run into the target zone 7 of the analysis device 1 without formation of a visible marking Irrespective of the presence of the analyte to be detected in the sample, either the analyte-specific binding partners which are conjugated with a substance or the substance itself binds to corresponding substance-specific binding partners in confectionery area 11, the substance thus bound with the dye components passing by or their precursors forms an insoluble or poorly soluble dye complex, which remains in place and by antibodies against the formed Dye complex, which are immobilized in the control field 11, are bound in large numbers with the duration of the test and form a clearly visible marking.

In control field 11 there are immobilized antibodies against the substance, e.g. β-galactosidase. As already mentioned, the reaction takes place in ConfroUfeld 11, regardless of the amount of analyte in the reagent.

The detection of an analyte with the enzyme ß-galactosidase as an enhancer system is carried out by the cleavage of natural or αβ-ß-D-galactosides in galactose and the corresponding residual compounds. Unphysiological substrates for the enzyme, such as ONPG or X-gal, give colored reaction products after hydrolysis and oxidation and allow visual and spectrophotometric detection.

Sfreptavidin and avidin, proteins with multiple binding sites for biotin, can be used as an alternative amplification system. For example, a specific binding partner can be labeled with biotin. Proteins such as sfreptavidin and avidin bind to one of their four binding sites in a highly specific manner to biotin. Free biotin binding sites are identified with the biotin-labeled enzyme, e.g. alkaline phosphatase, saturated. Due to the addition of chromogenic substrates, a color mixture precipitates, so that the binding can be verified. Since the biotin-binding proteins contain four biotin binding sites and the enzymes used can also ask several biotin groups, complexes can form with many protein-enzyme-biotin molecules, which significantly increases the sensitivity of detection. Alkaline phosphatase is used as a marker enzyme, since a sensitive, histochemical color reagent and signal amplifier system results in connection with suitable substrates. Substrates such as After hydrolytic cleavage by the enzyme, p-nitrophenyl phosphate gives colored, photometrically easily measurable reaction products or, as in the case of 5-bromo-4-chloro-3-indolyl phosphate (BCIP = X-phosphate), a deep blue colored insoluble, easily recognizable Precipitation of indigo. BCIP is used together with nitro blue tetrazohum (NBT) as a color enhancer.

The enzyme peroxidase (HRP) with derivatized tyramine can be used as a further alternative amplification system (Super-CARD, Catalytic deposition of derivatized tyramine, Bhattacharya, R., Bhattacharya, D., and Dhar, TK (1999), Journal of hnmunological Methods 227, 31-39). Here is z. B. the specific binding partner coupled with the enzyme peroxidase (HRP) and catalyzes with the help of H ₂ O ₂ (which is present in the reagent or is formed directly by a chemical reaction) the covalent binding of the derivatized tyramine to the analysis field 10 (specifically to specially modified proteins (p-OH-PPA-casein, p -OH-PPA-gelatin or p-OH-PPA-BSA; p-OH-PPA (3- (p-hydroxyphenyl) propionic acid)) which are immobilized on the analysis field 10). Either a dye, gold particle (NanoGold) or a catalyst is used as the molecule that is coupled to tyramine. In the latter case, for example peroxidase (HRP), a histochemical color reagent thus results in a signal which is amplified many times over.

The reagent used to take up the sample consists of monohydric alcohol with 1 to 5 carbon atoms, ketones with 3 to 8 carbon atoms, polyhydric alcohols, especially ethylene glycol and polyethylene glycol. The concentrations of these chemical compounds are selected from a range with a lower limit of 0.1%, in particular 5%, preferably 10%, and an upper limit of 40%, in particular 30%, preferably 20%.

A detergent, in particular (octylphenoxyl) polyethoxyethanol, alkylphenol polyglycol ether, Tween 20, sodium deoxycholate, nonidet P-40 (Ige-pal CA-630), Triton X-100, cholic acid, deoxycholic acid and / or Zwittergent® in a concentration selected from a range with a lower limit of 0.001%, in particular 0.005%, preferably 0.01%, and an upper limit of 1%, in particular 0.5%, preferably 0.2%, as a solubilizer become.

A buffer can also be added to the reagent, which keeps the pH of the reagent constant. Buffer solutions such as citrate buffer, acetate buffer, maleate buffer, phosphate buffer, colhdin buffer, triethanolamine-HCl-EDTA buffer, tris buffer, ammonium diol buffer, glycine buffer, diethanolamine buffer or tris-boric acid-EDTA buffer, or buffers according to Good, NE et al. (1966) Biochemistry 5, 467. The concentration of the buffer solutions is in each case selected in percent by weight from a range with a lower limit of 0.1%, preferably 1% and in particular 2% and with an upper limit of 20%, preferably 15%, in particular 10%. The buffer solution has a pH value selected from a range with a lower limit of 5.5, in particular 6.0, preferably 6.5, and an upper limit of 9.5, in particular 9.0, preferably 8.5. Buffer solutions with a pH value with a lower limit of 6.8, preferably 7.0, especially 7.2 and an upper limit of 8.0, preferably 7.8, in particular 7.6.

The addition of a mixture of the buffers mentioned makes it possible to stabilize the pH of the reagent over a wide range and thus to ensure a correct analysis result even when examining contaminated samples. Contamination from the existing buffer system is also buffered to such an extent that it does not interfere with the analysis reaction of the at least one analyte.

Furthermore, at least one dye component or its precursor can be added to the reagent, the dye components being selected from a group comprising tyramine and its derivatives, X-gal (5-bromo-4-chloro-3-hιdolyl-ß-D- Galactoside), S-Gal (3,4 cyclohexenoesculetin-ß-D-galactopyranoside), ONPG (o-nifrophenyl-ß-D-galactopyranoside), CPRG (chlorophenol red-ß-D-galactopyranoside), Bluogal (5- Bromo-3-h dolyl-ß-D-galactoside), MUGal (4-methylumbelhferyl-ß-D-galactopyranoside), NBT (nitroblue tetrazolium chloride), BQLP (5-bromo-4-chloro-3-hιdolylphosphate), PNPP (p-nitrophenyl phosphate), OPD (o-phenylenediamine), ABTS (2,2'-azino-di- (3-ethylbenz-tl iazoline Sulfonic Acid), DAB (3,3-diaminobenzidine), TMB (3.3 '5,5'-teframethylbenzidine), phenazine methosulfate, potassium ferrocyanide, potassium ferricyanide, ferric ammonium cifrate, VectorBlackTM, cherylluminescent dyes, such as Galacton-Star substrate, Lumi-PhosTM plus, LuciGLOTM, DuoLuXTM, Lumigen PS-3 HRP, cheimlumines (horseradish peroxidase) substrate, 2 component system from BioFXTM, and / or Lumi-Gal 530 and / or fluorescent dyes, e.g. 3-Carboxyumbelhfeιyl-ß-D-Ga-lactopyranosid (CUG) and / or MUP (4-methylumbelhferyl phosphate). The concentration of the dye components is selected from a range with a lower limit of 1 μM, in particular 10 μM, preferably 50 μM, and an upper limit of 300 mM, in particular 150 mM, preferably 100 mM.

The dye components and / or their precursors can be in undissolved form in the reagent or partially in the reagent or completely or partially in the dye field 12 or completely or partially in the starting zone 6 of the analysis device 1.

The dye components and / or their precursors are dissolved in the reagent and are released with a time delay by the addition of the components of the reagent, BSA, maltodextrin, milk powder, calf serum, casein and / or gelatin and transported via the analytical device 5 by the capillary action. In order to increase the osmolarity of the reagent, sodium salts, potassium salts, phosphate salts, magnesium and / or manganese salts, BSA, maltodextrin (M trin) or milk powder in a concentration from a range with a lower limit of 0.001%, in particular 0.1% , preferably 1%, and an upper limit of 30%, in particular 20%, preferably 10%.

Furthermore, a preservative, in particular sodium azide, sodium benzoate, sorbic acid, pentachlorophenol, sorbate and / or preservative based on mercury, can be added to the reagent in a concentration from a range with a lower limit of 0.01%, in particular 0.05%, preferably 0 , 1%, and an upper limit of 5%, in particular 3%, preferably 1%, in order to extend the shelf life of the reagent.

The following two exemplary embodiments show possible implementation options of the method, it being noted at this point that the invention is not limited to the specifically mentioned exemplary embodiments, but rather all conceivable exemplary embodiments which are included in the scope of protection by combining individual details of the described options.

Example 1:

Analysis device 1 for determining benzodiazepines in saliva

The sample in which the analyte to be determined is located is obtained from the saliva of a subject using a cotton swab. The stick is inserted into the analyte chamber 14 of the reagent container 2. Due to the action of a small force, such as lightly compressing the reagent container 2 with the container 3, the reagent chamber 13 and the analyte chamber 14 are pierced by the penetration device 4, such as a needle, and a connection is established, as a result of which the reagent passes over the sample of the cotton swab in the analyte chamber 14 flows to the start zone 6 or is drawn.

The start and finish zone 6, 7 is made of cellulose absorbent paper (Pall Coφoration, type 165 for the start zone 6 (BSP165PK) and type 197 for the target zone 7 (BSP197PK)). The format of the start and finish zone 6, 7 is 10 x 70 mm. The Filteφapier is used without further impregnation for the production of the analysis device 1. The dye field 12 is made of glass fiber microfilter type GF / D (Whatman). The format of the dye field 12 is 5 x 8 mm and is evenly charged with 15 μl of the dye solution. This is followed by drying with exclusion of light at 20 ° C. The dye solution consists of: X-Gal (100 mM), NBT (50 mM), Phenazine Methosulfate (1 mM), BSA (4%), Maltrin (1%) in PBS (5 mM KH ₂ PO ₄ , 15 mM Na ₂ HPO ₄ , 120 mM NaCl, 2.3 mM KC1, 2 mM MgCl ₂ , 0.05% NaN ₃ , 5% ethanol, 0.1% Triton X-100). The starting reagents are obtained from SIGMA-Aldrich.

The conjugate field 9 consists of a blocked Accuwick® membrane (Pall, AW 14-20-10). The membrane is blocked with 1% BSA in PBS solution for 1 hour at room temperature, then drying at 20 ° C. The shape is 5x 8 mm. 15 μl of the AntikÖiper solution are pipetted on evenly and then drying is carried out at 20 ° C. The antibody solution consists of: primary monoclonal antibodies (host mouse) against benzodiazepines (25 nM), (Fitzgerald, cat. No .: 10-B12) secondary antibodies anti-mouse (25nM) conjugated with beta-lactosidase (from Fisher Scientific) β-galactosidase (25 nM), (Röche Apphed Science), BSA (4%), maltrin (1%) in PBS (5 mM KH ₂ PO ₄ , 15 mM Na ₂ HPO ₄ , 120 mM NaCl, 2.3 mM KCL, 2mM MgCl ₂ , 0.05% NaN ₃ , 0.1% Triton X-100).

For the production of the running fields 8, S-tires made of Predator ™ (PALL) in the format 5 x 8 mm are used. To block binding sites, the Sfreifen in a 1% BSA-containing PBS solution (5 mM KH ₂ PO ₄ , 15 mM Na ₂ HPO ₄ , 120 mM NaCl, 2.3 mM KC1, 2 mM MgCl ₂ , 0.05 % NaN ₃ , 0.1% Tifron X-100) incubated for 1 hour at room temperature with shaking.

In the test field, a BSA-benzodiazepine conjugate (from Fitzgerald) is immobilized before the binding sites are blocked. To do this, the cellulose acetate filter (AcetatePlus) is incubated in 100 mM sodium periodate (SIGMA-Aldrich) for 20 min, then washed with H ₂ O and washed with a 0.5 μM BSA-benzodiazepine solution in 0.1 M borate buffer ( pH 9.0) for 10 minutes. Then 4 mg sodium cyanoborohydride (SIGMA-Aldrich) are added per ml and incubated for 2 h at room temperature. Unbound BSA benzodiazepine is washed away using H ₂ O.

In KonfroUfeld 11, a specific polyclonal antibody against the enzyme ß-galactosidase (Fitzgerald, host: Rabbit) is immobilized before the binding sites are blocked. The hnmobihsie- 0.25 μM specific polyclonal antibody is produced using the same method as in the test field.

The reagent used, in which the sample is taken up, consists of PBS (5 mM KH ₂ PO ₄ , 15 mM Na ₂ HPO ₄ , 120 M NaCl, 2.3 mM KCl, 2 mM MgCl ₂ , 0.1% BSA, 0.1% maltrine, 0.05% NaN ₃ , 5% ethanol, 0.05% Triton X-100) with a pH of 7.1.

The start and finish zones 6, 7 and the other fields are attached to Xeroperm (Rank Xerox Limited) and in container 3 by means of double-sided adhesive film (Tesa AG). The strips are stored at room temperature in the absence of light and moisture.

Embodiment 2

Analysis device 1 for the determination of testosterone in saliva

The sample in which the analyte to be determined is located is obtained from the saliva of a sample using a cotton swab. The stick is inserted into the analyte chamber 14 of the reagent container 2. Due to the action of a small force, such as lightly squeezing the reagent container 2 with the container 3, the reagent chamber 13 at the predetermined breaking point and the analyte chamber 14 are pierced by the penetration device 4, such as the start zone 6, and a connection is established, as a result of which the reagent passes through the Sample of the cotton swab in the analyte chamber 14 to the starting zone 6 flow or is drawn.

The start and finish zones 6, 7 are made of cellulose absorbent paper (Pall Coφoration, type 165 for the application pad (BSP165PK) and type 197 for the receiving pad (BSP197PK)). Sizes of 20 x 100 mm are cut which are used without further impregnation or pretreatment for the production of the analysis device 1.

The dye field 12 is made from 'Glass Fiber Media' (Pall, A / D Glass). Strips in the format of 5 x 80 mm are cut and evenly charged with 80 μl of the dye solution. The drying then takes place at 20 ° C in the absence of light. The dye solution consists of: X-Gal (100 mM), NBT (50 mM), Phenazme methosulfate (1 mM), BSA (4%), Maltrin (1%) in PBS (5 mM KH ₂ PO ₄ , 15 mM Na ₂ HPO ₄ , 120 mM NaCl, 2.3 mM KCl, 2 mM MgCl ₂ , 0.05% NaN ₃ , 5% ethanol, 0.1% Triton X-100) (all solutions are from SIGMA-Aldrich ). The running, analysis and confection area 11 consist of a 5 μm hnmunodyne® ABC membrane in the format 20 x 80 mm (Pall., BC500H5R). A 0.5 μM testosterone-3-CMO-BSA solution (from Fitzgerald, 80-TT49) is placed on the analysis device 1 at the location of the analysis field 10 and a 0.25 μM biotin-BSA solution at the location of the control field 11 ( SIGMA-Aldrich) applied. The remaining binding sites on the membrane are then blocked for 2 hours at room temperature with a 1% BSA in PBS solution.

In place of the conjugate field 9 on the analysis device 1, 10 nM of the primary specific monoclonal antibody against testosterone (host: mouse, Fitzgerald: 10-T07), 25 nM biotinylated anti-mouse IgG antibody (host: horse, Vector Laboratories: BA- 2080) and 100 nM of an avidin-beta-galactosidase conjugate (SIGMA-Aldrich) applied directly to the blocked hnmunodyne® ABC membrane between start zone 6 and analysis field 10.

The reagent used, in which the sample is taken up, consists of PBS (5 mM KH ₂ PO ₄ , 15 mM Na ₂ HPO ₄ , 120 mM NaCl, 2.3 mM KCl, 2 mM MgCl ₂ , 0.1% BSA, 0.1% maltrine, 0.05% NaN ₃ , 5% ethanol, 0.05% Triton X-100) with a pH of 7.1.

The cushions and the fitted hnmunodyne® ABC membrane are cut according to the specifications described and attached to the carrier material (Xeroperm type 003R96094 from Rank Xerox Limited) using double-sided adhesive film. Of these, strips of 4 mm are cut with a roll cutter. The strips are fastened in the container 3 of the analysis device 1.

The analysis device 1 is stored at room temperature with the exclusion of light and moisture.

Finally, it should also be mentioned that in particular the antibodies which are present in the antibody solution are each adapted to the analyte to be detected.

The exemplary embodiments show possible design variants of the analysis device 1, it being noted at this point that the invention is not restricted to the specifically illustrated design variants of the same, but rather also various combinations of the individual design variants with one another are possible and this variation possibility is based on the teaching of the technical Acting through the subject invention lies in the ability of the person skilled in the art in this technical field. So all conceivable leadership variants, which are possible by combining individual details of the illustrated and described variant, included in the scope of protection.

For the sake of order, it should finally be pointed out that, for a better understanding of the structure of the analysis device 1, these or their components have been partially shown to scale and / or enlarged and / or reduced.

The object on which the independent inventive solutions are based can be found in the description.

Above all, the individual versions shown in FIGS. 1 to 4 can form the subject of independent solutions according to the invention. The relevant tasks and solutions according to the invention can be found in the detailed descriptions of these figures.

REFERENCE NUMBERS

Analysis device Reagent container Container Penefrationseinrichtung Analysis section Start zone Target zone Running field Conjugate field Analysis field KonfroUfeld Dye field Reagent chamber Analyzer chamber Carrier Closure element Flap Positional control element

Claims

Patent claims

1. Analysis device (1) for determining at least one analyte from a sample comprising a reagent container (2) and a container (3), in which an analyzer (5) made of capillary material is arranged, and by a penetration device (4), such as eg An open end of a capillary is connected to the flow, characterized in that at least one reagent chamber (13) for the reagent and / or at least one analyte chamber (14) for receiving the analyte are arranged in a flow-connectable manner in the reagent container (2).

2. Analysis device (1) according to claim 1, characterized in that a means for connecting the reagent container (2) to the container (3), such as. a tongue and groove connection, a thread, a bayonet connection, a predetermined breaking point, a folding hinge is arranged.

3. Analysis device (1) according to one of claims 1 or 2, characterized in that one or more penetration device (s) (4) is (are) designed as a cannula, tip, wick, spike, etc.

4. Analysis device (1) according to one of the preceding claims, characterized in that a wall of the reagent container (2), the reagent chamber (13), the analyte chamber (14) and / or the container (3) is at least partially formed by a septum ,

5. Analysis device (1) according to claim 4, characterized in that in the wall of the reagent container (2), the reagent chamber (13), the analyte chamber (14) and / or the container (3) a predetermined breaking point for transferring the reagent into the Analyte chamber (14) and / or the container (3) is formed.

6. Analysis device (1) according to one of the preceding claims, characterized in that at least one positionable closure element (16) is arranged on the reagent container.

7. Analysis device (1) according to claim 6, characterized in that at least two closure elements (16) are arranged in parallel.

8. Analysis device (1) according to one of the preceding claims, characterized in that a flap (17) which is pivotable by 90 ° is arranged in the analyte chamber (14).

9. Analysis device (1) according to one of the preceding claims, characterized in that the reagent container (2) on the container (3) is movable, such as. is arranged to be displaceable, rotatable and / or foldable.

10. Analysis device (1) according to one of the preceding claims, characterized in that in the reagent container (2) and / or container (3) a position control (18) for checking the relative movement, such as e.g. the sliding, rotating and / or folding movement. supply, the container (3) and the reagent container (2) is formed.

11. Analysis device (1) according to one of the preceding claims, characterized in that one or more analysis sections (5) are arranged in the container (3) for each different analyte.

12. Analysis device (1) according to one of the preceding claims, characterized in that the analysis section (5) is formed from an absorbent material.

13. Analysis device (1) according to claim 11 or 12, characterized in that the analysis section (5) consists of at least one analysis field (10).

14. Analysis device (1) according to one of the preceding claims, characterized in that one or more strips of absorbent material are arranged in the container (3) at at least one end of the analysis section (5).

15. Analysis device (1) according to claim 14, characterized in that the Sfreifen is arranged at two diametrically lying ends of the Analytesfrecke (5).

16. Analysis device (1) according to one of claims 14 or 15, characterized in that the tire is designed as a penetration device (4).

17. Analysis device (1) according to one of claims 14 to 16, characterized in that a recess is formed above the tire.

18. Analysis device (1) according to one of claims 11 to 13, characterized in that the analytical device (5) in a starting zone (6) and at least one running, dye, conjugate, control and / or analysis field (8, 12, 9, 11, 10) is divided.

19. Analysis device (1) according to claim 18, characterized in that in the analysis field (10) at least one analyte and / or an analyte-specific binding partner is immobilized in a predefined concentration.

20. Analysis device (1) according to claim 18 or 19, characterized in that at least the analysis field (10) of the analysis section (5) in the container (3) is arranged visibly.

21. Analysis device (1) according to one of the preceding claims, characterized in that one or more markings are arranged on the container (3) to support the evaluation of the analysis.

22. Analysis device (1) according to one of the preceding claims, characterized in that a fixing device (19), such as e.g. a snap-in device, which fastens the reagent container (2) in a predefined position on the container (3), is arranged on the container (3) and / or on the reagent container (2).

23. Analysis device (1) according to one of the preceding claims, characterized in that one or more auxiliary substances for carrying out the analysis, such as stabilizers, preservatives and or drying agents, are present in the container (3).

24. Analysis device (1) according to one of the preceding claims, characterized in that in the analyte chamber (14) an analyte from a group comprising active substances, honnones, proteins, peptides, allergens, antigens, antibodies, neurofransmitters, nucleic acids, carbohydrates and / or Lipids is arranged.

25. Analysis device (1) according to claim 24, characterized in that active substances from a group comprising drugs, drug substitutes, performance-enhancing substances such as doping agents, drugs, toxins or their metabolites are arranged.

26. Method for determining at least one analyte from a sample with an analysis device (1) consisting of a reagent container (2) and a container (3), in which at least one analyzer device (5) made of capillary-capable material is arranged, which is brought into contact with one another are characterized in that the reagent container (2) and the container (3), which are already arranged one another, are fluidly connected by means of a permeability device (4), the reagent container (2) and the container (3) being moved relative to one another, such as eg moved, folded or rotated.

27. Use of the analysis device (1) according to one of claims 1 to 25, characterized in that at least one analyte from a group comprising active ingredients, hormones mones, proteins, peptides, allergens, antigens, antibodies, neurofransmitters, nucleic acids, carbohydrates and / or lipids is detected.