JP4451735B2 - Improved detection device - Google Patents

Description

  The present invention relates to a flow-through detection apparatus including a solid phase support in an immunoassay method for detecting an antigen or an antibody, comprising a flow rate control and / or concentration means capable of controlling a flow rate and concentrating a reagent and a sample. A flow rate control and / or concentration means that can control the flow rate and concentrate the reagent and sample, and coloration / non-coloration of the test region where the concentration means is a capture reagent site and a control site Non-test region other than the capture reagent site and the control site The present invention relates to a flow-through detection device having a function that can be confirmed before the end of the test by comparison with the above, an immunoassay method and an immunoassay kit using the same.

  Various methodologies such as an immunodiffusion method, an enzyme immunoassay method, and an agglutination method have been put to practical use as analytical methods for detecting or quantifying an analyte in a sample by an immunological technique using the specificity of the immune reaction. The flow-through inspection method and the immunochromatographic inspection method (lateral flow method, tangential flow method) are inspection methods using a membrane, are easy to operate, and are widely used in general inspection places. Various principles of the lateral flow inspection method have been reported (see Non-Patent Document 1 and Patent Documents 1 to 9). Various principles of the flow-through inspection method have also been reported (see Non-Patent Document 1 and Patent Documents 10 to 15).

  This analysis method will be briefly described by taking detection of influenza virus antigens as an example. Specimens collected from a patient (pharyngeal / nasal wiping solution, nasal aspirate, etc.) are suspended in a sample suspension on a membrane on which a capture reagent (eg, anti-influenza virus antibody) for capturing influenza virus antigens is immobilized. When a predetermined amount of the sample is dropped, when the sample liquid passes through the membrane, the existing analyte (influenza virus antigen) is captured by the capture reagent immobilized on the membrane. Next, when a predetermined amount of a detection reagent (for example, a labeled antibody) that is labeled with an insoluble colored particle such as an enzyme or colloidal gold particles and binds to the analyte is dropped, a capture reagent-analyte-detection reagent ( A labeled antibody) is formed. Thereafter, in the case of the enzyme-labeled antibody, an immune complex was formed by dropping the substrate, and in the case of using insoluble colored particles, it was concentrated on the immobilized capture reagent and visualized. Visually determine the presence of influenza virus antigen in the sample by coloring the area and comparing the color of the colored test area (capture reagent site, control site) and the other uncolored non-test area be able to. This analysis method is widely used because it has high sensitivity and does not require special equipment and equipment, and can easily and quickly obtain results.

  In the flow-through type detection device, the sample is not added directly onto the membrane, but is added to a container in which the sample can be temporarily stored, the complex of the analyte and the labeling reagent is concentrated in the container, and the sample The flow rate when the sample is moved from the container to the membrane and absorbed is controlled by the shape and size of the opening through which the sample moves or by an appropriate filter. And / or some have a concentration means.

  However, among commercially available immunoassay methods using flow-through detection devices, products having flow rate control and / or concentration means (test pack RSV, Directigen, etc.) have their flow rate control and / or concentration means ( In most cases, it is difficult to make the determination until the adapter is removed, or a multi-step operation step is required even after the flow rate control and / or concentration means is removed. On the other hand, although flow-through detection devices that do not have flow rate control and / or concentration means are available on the market (Morinaga's pregnancy diagnostic agents, overseas HCV diagnostic agents), these products have an effective concentration effect on samples and reagents. Not only can this be expected, but also the absorption time control of these liquid components depends only on the membrane that is the solid support and the porous band that accompanies the absorption band, so sufficient performance (detection sensitivity) depending on the measurement target There was a case that could not be achieved.

  In addition, an improved flow-through type detection device that has flow control and / or concentration means and improved operability is available (Quick S-Flu A / B "Seiken"). Because of the presence of the concentration means, the test area could not be visually observed in the middle of the measurement, and it was difficult to confirm and determine the progress until the sample / reagent had completely absorbed.

Japanese Patent No. 1774328 Japanese Patent No. 1889525 Japanese Patent No. 2590059 Japanese Patent No. 2058991 Japanese Patent No. 2131938 Japanese Patent No. 3304350 Japanese Patent No. 1963966 Japanese Patent No. 2133513 Japanese Patent No. 2135865 Japanese Patent No. 3114531 Japanese Patent No. 1879967 Japanese Patent No. 2818191 Japanese Patent No. 2035586 JP 57-200862 A JP 59-170768 Guide to Diagnostic Rapid Test Device Components ", 2 nd edition, published by Schleicher & Schuellcompany, January 2000, Edited by Lisa Vickers, p6- 8

  Therefore, the present invention is an immunoassay using a flow-through detection device having a flow rate control and / or concentration means capable of concentrating reagents and samples by controlling the flow rate, while maintaining the sample / reagent concentration effect, An object of the present invention is to provide an immunodiagnostic apparatus capable of observing the progress of the test and a kit using the same.

  In the flow-through type detection apparatus having flow control and / or concentration means capable of concentrating the reagent and sample by controlling the flow rate, the inventor uses the flow control and / or concentration means as a test region containing a capture reagent and the test region. The present invention has been completed by finding that the above-mentioned problems can be achieved by molding from a transparent resin that can be seen through the solid support, or a resin having a color that matches the solid support that includes the test region. . That is, the present invention relates to a flow-through type detection device that combines a flow rate control and / or concentration means molded from a transparent resin or a solid phase support and a resin of the same color.

  The present invention controls the flow rate of the above resin consisting of ABS (acrylbutyl styrene), PE (polyester), PP (polypropylene), PS (polystyrene), PC (polycarbonate), PA (polyallomer), and concentrates the reagent and sample. The present invention relates to a flow-through type detection device that combines a flow rate control and / or concentration means that can be used.

  The present invention further provides a flow rate control capable of concentrating a reagent and a sample by controlling a flow rate in which the color of the resin is a color (for example, a white nitrocellulose membrane filter) synchronized with a filter membrane which is a solid phase support including a test region. The present invention relates to a flow-through detection device combined with a concentration means.

Specifically, the present invention is as follows.
[1] A solid phase support on which a capture reagent that specifically binds to an analyte in a specimen sample is immobilized and a labeling reagent that includes a ligand that specifically binds to the analyte, and the reagent and the sample are in a solid phase A flow-through detection device for performing an immunoassay having a flow rate control and / or concentration means capable of concentrating a reagent and a sample by controlling a flow rate moving to a support, and detecting the flow rate control and / or concentration means A flow-through type detection device that can confirm the coloring or non-coloring of the test area without removing it from the device,
[2] The flow-through detection device according to [1], wherein the labeling reagent is contained in the porous material, and the porous material is provided in the flow rate control and / or concentration means,
[3] The flow-through detection device according to [1], wherein the labeling reagent is contained in a porous material and includes a sample addition device equipped with the porous material containing the labeling reagent,
[4] The color of the test area and the non-test area on the solid support can be simultaneously observed without removing the flow rate control and / or concentration means from the detection device, and the test area can be colored or non-colored. The flow-through detection device according to any one of [1] to [3], which can be confirmed by comparison with
[5] The flow-through detection device according to any one of [1] to [4], wherein the flow rate control and / or concentration means is integrated with the detection device.
[6] The flow-through detection device according to any one of [1] to [5], wherein the flow rate control and / or concentration means is molded from a transparent resin.
[7] The flow-through detection device according to any one of [1] to [5], wherein the flow rate control and / or concentration means is molded from a resin of a color synchronized with the solid support.
[8] The flow-through detection device according to any one of [1] to [7], wherein the flow rate control and / or concentration means is molded from polystyrene resin.
[9] The flow-through detection device is composed of a resin main body containing a solid support and a resin flow control and / or concentration means, and the main body and flow control and / or concentration means are made of polystyrene resin. The flow-through detection device according to any one of [1] to [8] to be molded,
[10] The flow-through detection device according to [9], wherein the flexural modulus of polystyrene resin molding the flow rate control and / or concentration means is different from the flexural modulus of polystyrene resin of the main body by 200 MPa or more.
[11] The flow-through detection device according to [9], wherein the rigidity of the polystyrene resin molding the flow rate control and / or concentration means is lower than the rigidity of the polystyrene resin of the device body.
[12] The flow-through detection device according to any one of [1] to [11], wherein a distance between the side of the reagent and sample addition unit of the flow rate control and / or concentration means and the outer structure is 1 mm or more.
[13] The flow-through detection device according to any one of [1] to [12], wherein the color of the outer structure on the side of the reagent and sample addition unit of the flow rate control and / or concentration means is a light color,
[14] There is no condition in which light having a specific wavelength is intensified by interference between light reflected by the flow rate control and / or the lower surface of the concentration unit and the reflected light on the upper side of the flow rate control and / or the concentration unit after addition of the reagent and the sample The flow-through detection device according to any one of [1] to [13],
[15] The flow-through of any one of [1] to [14], after the reagent and sample are added, the lower surface of the flow rate control and / or concentration means and the solid support are in close contact with each other without forming an air layer between them. Type detection device,
[16] If the cross-sectional shape of the flow rate control and / or concentration means reagent and sample addition part does not include a part of the parabola or includes a part of the parabola, the focus is not in the test region [1] to [ 15], and
[17] The flow-through detection device according to any one of [1] to [16], wherein the flow rate control and / or concentration means is provided with a color for determining positive or negative or comparing and determining the degree of positive.

  In the present invention, the flow rate control and / or concentration means capable of concentrating the reagent and the sample by controlling the flow rate is a transparent region that can be seen through the test region including the capture reagent and the control region and the solid phase support including the test region. It is molded from a resin or a resin of the same color as the solid support containing the test area. By using a transparent resin, it is possible to recognize the coloration of the capture reagent site through flow control and / or concentration means. Further, by using a resin having a synchronized color, recognition of the colored site of the capture reagent site and the control site is not hindered by the color of the flow control and / or concentration means. Therefore, in the apparatus of the present invention, it is possible to make the determination without hindering the color determination of the capture reagent site without removing the flow rate control and / or the concentration means. For example, when normal detection is performed, the detection time is specified, and after the detection time has elapsed, the flow rate control and / or the concentration means is removed and the color determination is performed. On the other hand, when detecting using the apparatus of the present invention, for example, if the sample is a strong positive sample, it is known that the sample is positive before the specified detection time elapses. Can be completed and the time required for detection can be saved. In addition, when detecting an infectious disease, bacteria or viruses may be present in the specimen. In such cases, it is desirable not to touch the flow control and / or concentration means to reduce the risk of infection. When the apparatus of the present invention is used, it is not necessary to touch the flow rate control and / or concentration means to which the sample is added, so that the risk of infection can be reduced.

Hereinafter, the present invention will be described in detail.
The present invention relates to an apparatus for detecting an analyte in a sample, and relates to a flow-through detection apparatus and a method for detecting an analyte using the apparatus. The flow-through detection apparatus includes a membrane-like solid support on which a capture substance capable of binding and capturing at least an analyte is immobilized. In the flow-through detection method, a sample containing an analyte is first sampled on a membrane (solid support) on which a capture reagent (eg, antibody) for capturing the analyte (eg, antigen) is immobilized. When a predetermined amount of the sample suspended in the suspension is supplied, the analyte present when the sample passes through the solid support is captured by the capture reagent immobilized on the solid support, and the analyte-capture A reagent complex is formed. Next, when a predetermined amount of a labeling reagent containing a ligand that specifically binds to the analyte (eg, enzyme label for the analyte, insoluble particulate matter) is provided, the capture reagent-analyte-labeling reagent complex is captured. It is formed at the reagent immobilization position (test area). Then, the labeling reagent can be added by any method (in the case of enzyme labeling, a substrate is added to cause a color reaction, and in the case of insoluble particulate matter labeling, insoluble particulate matter accumulates on the capture reagent and the color of the particulate matter can be recognized. ) To determine the presence of the analyte. A site where the capture reagent is immobilized is called a capture reagent site. In the flow-through detection method, the specimen sample passes across the solid support. The flow-through detection device may include a control reagent site on which a control reagent is immobilized. A test region refers to a region that can be colored on a solid support and includes a capture reagent site and a control reagent site. The non-test region refers to a region that is not colored, and includes sites other than the capture reagent site and the control reagent site. By comparing the colors in the test area and the non-test area, it is possible to visually recognize whether or not the area is colored. In this case, the test area and the non-test area are visually observed at the same time, and the determination is facilitated by comparing both colors.

  The flow-through detection device of the present invention is a device having flow rate control and / or concentration means for specimen samples and reagents. FIG. 1 shows the configuration of a flow-through type detection apparatus having flow control and / or concentration means, and an outline of the method of the present invention using the apparatus. The flow-through detection device of the present invention comprises a flow rate control and / or concentration means capable of concentrating reagents and samples by controlling the flow rate with the main body, and by fitting the flow rate control and / or concentration means with the main body, etc. Combined and used. Although there are no restrictions on the material, shape, etc. of the main body, a membrane-like solid phase support in which a capture substance capable of binding and capturing at least an analyte is fixed in a resin container (housing). The resin housing and the resin flow control and / or concentration means are fitted and used. For example, as shown in FIG. 1, a solid support is housed in a housing. There is an opening in the upper part of the housing, and the test area of the solid support is exposed in the opening. A part of the flow control and / or concentration means is coupled so as to be close to or in contact with the opening.

  Further, the flow rate control and / or concentration means for the sample and reagent of the flow-through detection device of the present invention may be mounted in advance on the flow-through detection device and integrated with the detection device body. Here, “integrated with the detection device main body” means that the flow rate control and / or concentration means is connected to the device main body in a non-removable or difficult to remove manner. In the apparatus of the present invention, since the result can be determined without removing the flow control and / or concentration means, it is necessary to remove the flow control and / or concentration means when determining the result as in the conventional apparatus. There is no. For example, the flow rate control and / or concentration means may be integrally molded with the apparatus main body, or may be separately molded or difficultly coupled using a coupling means such as an adhesive after being molded separately. It is difficult to remove the fitting part of the flow rate control and / or concentration means and the apparatus main body so that the strength of the fitting when the fitting part of the flow rate control and / or concentration means is fitted, for example, is increased. You may do it. Here, difficult to remove means that it is not assumed to be removed in a normal use state, does not come off naturally, and requires a large force to be removed.

  Here, the flow rate control and / or concentration means capable of controlling the flow rate and concentrating the reagent and the sample controls the flow rate when the specimen sample and the reagent supplied to the solid phase support flow to the solid phase support, and A means for concentrating the specimen sample and the reagent flowing on the solid support to enhance the formation efficiency of the capture reagent-analyte-labeling reagent complex on the capture reagent on the solid support. Say. The flow rate control and / or concentration means comprises a member having a flow rate control function of the specimen sample and / or a reagent and sample concentration function. Examples of the member having such a function include a container-like adapter to which a specimen sample can be added and the specimen sample can be temporarily stored. In the flow-through detection device, the adapter is provided so as to be positioned above the solid support to which the capture reagent and the like are immobilized. There are moving openings. By changing the shape and size of the opening, the flow rate and speed when the specimen sample and reagent in the container move to the solid support can be adjusted and controlled. The shape and size of the opening are not limited, but can be appropriately designed according to the analyte, desired sensitivity, desired detection time, and the like. A filter may be disposed between the container and the solid support. The filter is not limited as long as it can remove impurities such as aggregates and solids contained in the specimen. Paper filter paper, glass filter, membrane filter having a pore size of 2 μm or less (for example, 0.22 μm, 0.45 μm) Etc. can be used. The filter can also control the rate at which the specimen sample and reagent in the container move to the membrane. Also in this case, the speed at which the specimen sample and the reagent in the container move to the solid support can be controlled by changing the thickness of the filter, the size of the filter hole, and the like. In this case, the container-like adapter and the filter are integrated to form a flow rate control and / or concentration means. By controlling the speed at which the specimen sample and reagent in the container move to the solid support, the time for the specimen sample to be absorbed by the solid support is controlled, and the capture reagent on the solid support per unit time It is possible to control the amount of the analyte in the sample sample that comes into contact with the sample, and the same effect as that obtained by concentrating the reagent can be obtained. Further, the sample and the labeling reagent may be mixed in the concentration means, and the labeling reagent may be previously contained in the container. For example, a labeling reagent may be included and equipped in a container adapter that can add a specimen sample and temporarily store the specimen sample. At this time, the apparatus may be equipped with a labeling reagent in advance, or may be equipped with a labeling reagent at the time of use. The labeling reagent may be liquid or freeze-dried, and is absorbed into a suitable porous material such as a glass fiber nonwoven fabric, dried, and a section (labeling reagent pad) cut from the porous material is placed. Also good. In this case, since the sample sample and the labeling reagent are mixed and reacted in the container, the complex of the analyte and the labeling reagent in the sample sample is concentrated when the sample sample moves from the container to the solid support. The The labeling reagent and the sample may be mixed, for example, in a sample addition device. The specimen addition device is an accessory part outside the detection apparatus, and is a container for putting a collected specimen into a specific process. Such devices include vials, syringes, tubes and the like. In addition to the container, the device may include a filtering means for filtering the specimen when the specimen is added to the detection apparatus. The sample addition device includes a container part for storing the sample and a part for adding and supplying the sample in the container to the detection apparatus. The latter part to which the specimen is added and supplied has a part having a nozzle (opening) for discharging the specimen from the container, and this part can also serve as a lid for the container part. The portion where the latter specimen is added and supplied to the detection device is also referred to as a nozzle portion or a lid portion. In addition, when the specimen addition device includes a filtering means, the specimen addition device may be composed of two parts, a nozzle portion including a filter housing including a filtration filter and a container portion. In this case, the filter housing has an opening through which the sample passes through the filter and an opening through which the sample that has passed through the filter is discharged, and the filter exists between both openings. As the container, for example, a syringe or the like that can pass the sample through the filter under pressure may be used. When the specimen addition device is used, the flow-through detection device of the present invention also includes the specimen addition device. That is, the flow-through detection apparatus of the present invention includes a labeling reagent, but the labeling reagent may be included in the apparatus or the sample addition device.

  In this way, the flow rate control and / or concentration means capable of controlling the flow rate and concentrating the reagent and sample in the apparatus of the present invention concentrates the complex of the analyte and the labeling reagent in the specimen sample, on the solid support. It is possible to control the rate of the complex of the analyte and the labeling reagent in the specimen sample that reaches the capture reagent.

  Flow rate control refers to keeping a mixture of a reagent and a sample for a certain period of time. In general, reagents and samples take a certain amount of time for the reaction, but if they flow to the solid support before the reaction has sufficiently progressed, the expected reaction will not occur on the solid support and the correct test results will be obtained. I can't. Therefore, by controlling the flow rate, the mixture of the reagent and the sample is kept only for the time required for the reaction between the reagent and the sample, and a correct test result can be obtained by sufficiently proceeding the reaction.

  Concentration means that a mixture of a reagent and a sample is passed only through a predetermined range of a solid support. In the flow-through type detection apparatus, a mixture of a reagent and a sample is passed through a solid support and the reaction is judged to determine the test result. In general, when the area of the solid support is large compared to the amount of the mixture of reagent and sample, a sufficient amount of analyte or analyte-capture reagent complex is supplied to the capture reagent on the solid support. Therefore, the detection sensitivity of the test result is lowered. Therefore, by concentrating the reagent and the sample, the mixture of the reagent and the sample can be passed only to a predetermined range of the solid phase support, and a sufficient amount of the analyte or analyte can be passed to the capture reagent on the solid phase support. Since the analyte-capture reagent complex is supplied, the detection sensitivity can be increased.

  In the present invention, since the flow rate is a flow rate per unit time, it may be referred to as a flow rate control means instead of the term “flow rate”. Furthermore, the flow rate control and / or concentration means of the present invention also has a function of controlling the flow path.

  In order to improve the absorption time adjustment of the analyte or reagent to the solid support and the concentration effect on the solid support, as a result, to improve the detection sensitivity of the analyte, the flow rate as described above As a flow-through detection device with flow control and / or concentration means that can control the concentration of reagents and samples, Becton Dickinson's Directive Series, Denka Seiken Influenza A-B-Quick "Seiken", Denka Seiken Quick S -Flu A / B "Seiken", Abbott Test Pack RSV, etc. In these conventional apparatuses having flow rate control and / or concentration means capable of controlling flow rate and concentrating reagents and samples, the components constituting flow rate control and / or concentration means are usually molded from non-transparent colored resin. Since the flow rate control and / or concentration means is provided on the solid phase support so as to cover the solid phase support on which the capture reagent is immobilized, in order to determine the detection result in the conventional apparatus, the flow rate is determined. It was necessary to remove the control and / or concentration means. There is also a device in which the capture reagent site exists in a shape adapted to the opening at the lower part of the flow rate control means, and in this case, it is possible to visually observe the color change or the like appearing in the capture reagent site through the opening. The contrast between the color of the flow rate control and / or concentration means and the color of the capture reagent site is unclear and it is difficult to make an accurate determination.

  In the present invention, the flow rate control and / or concentration means capable of concentrating the reagent and the sample by controlling the flow rate is formed of a transparent resin, or a resin of a color synchronized with the solid phase support including the test region It is molded from. By using a transparent resin, it is possible to recognize the coloration of the capture reagent site through flow control and / or concentration means. Further, by using a resin having a synchronized color, visual recognition of the colored portion of the capture reagent portion is not hindered by the color of the flow control and / or concentration means. Therefore, in the apparatus of the present invention, the detection result can be obtained without hindering the color determination of the capture reagent site without removing the flow rate control and / or concentration means. For example, when normal detection is performed, the detection time is specified, and after the detection time has elapsed, the flow rate control and / or the concentration means is removed and the color determination is performed. On the other hand, when detecting using the apparatus of the present invention, for example, if the sample is a strong positive sample, it is known that the sample is positive before the specified detection time elapses. Can be completed and the time required for detection can be saved. In addition, when detecting an infectious disease, bacteria or viruses may be present in the specimen. In such a case, in order to reduce the risk of infection, it is desirable not to touch the flow control and / or concentration means to which the sample is added. When the apparatus of the present invention is used, it is not necessary to touch the flow rate control and / or concentration means to which the sample is added, so that the risk of infection can be reduced.

  “Transparent” is one of the attributes of an object, which means that the object transmits radiation such as light, and is colored and colorless and transparent. In the present invention, colorless and transparent are desirable. Transparent resins include ABS (acryl butyl styrene), PE (polyester), PP (polypropylene), PS (polystyrene), PC (polycarbonate), PA (polyallomer), etc., and control the flow rate of the apparatus of the present invention. The flow rate control and / or concentration means capable of concentrating the reagent and sample can be configured by appropriately combining these resins.

  The degree of transparency of the flow rate control and / or concentration means is preferably 80% or more in terms of total light transmittance (2 mmt), but is not limited thereto. The total light transmittance can be measured, for example, according to ASTM-D1003 (American Society of Testing Material standard of American Material Testing Association) or JIS7105-1981.

  Conventionally, a high-impact general-purpose grade has been used as the resin for both the flow-through detection device main body and the flow rate control and / or concentration means capable of concentrating reagents and samples by controlling the flow rate. When assembled in this combination, the structure of the fitting portion of the main body and the flow rate control and / or concentration means (the claw structure on the main body side) may be damaged, and the function may not be performed (fitting is impaired). To solve this, the resin on the adapter side has a lower elastic modulus than the resin on the main body side (for example, the bending elastic modulus of the main body resin: 2350 MPa, the electrified ABSGR-2000, etc., the bending elastic modulus of the concentration mechanism resin: 1950 MPa, By using a material such as electrified transparent ABS (TE-30), it is possible to avoid damage to the structure of the fitting part. In addition, automatic assembly by a machine was difficult in the conventional situation (when a part is easily fitted by mechanical force, the claws are easily damaged), but by adopting this method, automation is possible. Thus, it is desirable that the resin used on the flow rate control and / or concentration means side has lower rigidity than the main body resin. For example, the flow rate control and / or the concentration means may have a low flexural modulus, but if it is too low, the fitting is lost due to the shrinkage of the resin during molding or the like, and 1500 MPa or more is necessary. The difference in flexural modulus between the resin of the flow rate control and / or concentration means and the resin of the main body is preferably 200 MPa or more.

  As described above, the color of the resin of the flow rate control and / or concentration means may be synchronized with a filter membrane such as a white nitrocellulose membrane filter that is a solid support serving as a test region. Here, “synchronization” means that the colors are similar. The most common color perceived when viewing an object is the surface color due to the reflected light from the surface of the object. There are chromatic colors such as red, yellow, and green, and achromatic colors such as white and black. Chromatic colors can be distinguished for several shades, such as red, yellow, and blue. This fundamental difference in hue determined according to the wavelength of light reaching the retina is called hue. In both chromatic and achromatic colors, when the brightness of light changes, the degree of brightness of the color also changes. This is called lightness. In addition, in chromatic colors, there are colors that are vivid and frightening, and colors that are astringent and cloudy depending on how the hue of the hue is included. This degree of purity of color is called chromatic value (saturation). These are the three basic properties of color and are called the three attributes of color. However, it does not strictly perceive colors according to the classification of three attributes. The impression of color is based on the nature of the color, including both brightness and saturation, which is the color tone. Here, the substantially synchronized color means a color having a color tone that has only a difference that cannot be recognized by human vision, and several panelists have a sensory test on whether or not it can be recognized as a different color. When done, it means a color that many panelists cannot recognize as a different color. For example, when white nitrocellulose membrane is used as the membrane and ABS resin is used as the flow control means, the color of the flow control means is, for example, cream (for example, UMG ABS color code B), beige (same E), white (same W) Alternatively, colors that are substantially synchronized with these may be employed. In the present invention, a capture reagent is immobilized on a membrane, and a complex of capture reagent-analyte-labeling reagent is formed on the reagent, so that the labeling reagent accumulates and the color of the labeling reagent is visualized. Can be recognized. As the membrane, for example, a white nitrocellulose membrane is used, and for example, a red gold colloid is used as a labeling substance of the labeling reagent. In this case, red gold colloid accumulates on the white membrane, so that the red color can be clearly recognized visually. The flow rate control and / or concentration means has an opening as shown in FIG. 1, and is arranged in the apparatus main body so that the opening and the capture reagent part overlap each other. When the flow control and / or concentration means is colored dark, the contrast between the color of the flow control and / or concentration means and the color of the capture reagent is clear even if the color of the capture reagent site is recognized through the opening. Therefore, the color depth cannot be accurately recognized. If the flow control and / or concentration means is removed, the color of the labeling substance can be recognized by the contrast between the color of the membrane and the color of the labeling substance. When the color of the flow control and / or concentration means is synchronized with the color of the membrane, the contrast between the color of the labeling reagent and the color of the flow control and / or concentration means that can be visually recognized through the opening of the flow control and / or concentration means The color of the labeling reagent can be recognized even when the flow rate control and / or concentration means is still installed in the apparatus main body. When a white nitrocellulose membrane is used as the membrane, the synchronized color is white or a color close to white, and examples thereof include light gray and cream. In this case, it does not matter whether it is transparent or opaque.

  The flow rate control and / or concentration means of the present invention is provided with a distance between the side of the reagent and sample addition part of the flow rate control and / or concentration means and the outer structure of 0.5 mm or more, preferably 1 mm or more. Light can enter the apparatus from the gap between the concentration means and the outer structure, and sufficient light can be supplied to the test area and / or the non-test area, so that the color determination can be accurately performed. Here, the reagent and sample addition part side of the flow rate control and / or concentration means refers to the outside of the part of the flow rate control and / or concentration means to which the container-like reagent and sample are added, and the outer structure means An apparatus main body portion to which the flow rate control and / or concentration means is mounted, and refers to a structure portion surrounding the flow rate control and / or concentration means. The positional relationship between the outer structure and the flow rate control and / or concentration means is as shown in FIG. In FIG. 2, the distance between the side of the reagent and sample addition portion of the flow control and / or concentration means and the outer structure means the distance between the upper end of the flow control and / or concentration means and the outer structure. . In FIG. 2, the light indicated by the arrows is shown entering through the gap between the upper end of the flow control and / or concentration means and the outer structure.

  FIG. 2 is an explanatory view of a cross section of the flow rate control and / or concentration means. By providing the flow control and / or concentration means 7 with a distance from the outer structure 10 of 0.5 mm or more, preferably 1 mm or more, it becomes possible to sufficiently supply the light 11 to the solid phase support 2 and accurately determine the color determination. It can be carried out.

  In addition, it is desirable that the color of the outer structure on the side part of the reagent and sample addition portion of the flow rate control and / or concentration means of the present invention is light. The light color referred to here means a light and light color such as white, cream and yellow. FIG. 3 is an explanatory view of a cross section of the flow rate control and / or concentration means. By making the color of the outer structure 10 on the side of the reagent and sample addition part of the flow rate control and / or concentration means light, the light 11 from the upper part is reflected by the outer structure and supplied to the solid support 2 sufficiently. This makes it possible to accurately determine the coloration.

  Further, when the flow control and / or concentration means of the present invention is molded from a transparent resin, the flow control and / or the reflected light on the lower surface of the concentration means and the reflected light on the upper surface of the flow control means after adding the mixture of the reagent and the sample. It is desirable not to be in a condition where light of a specific wavelength is intensified by light interference due to. If interference occurs between the reflected light on the lower surface of the flow rate control and / or concentration means and the reflected light on the upper surface of the flow rate control and / or concentration means, a specific color develops and the color tone of the non-test area cannot be accurately confirmed. It is.

  FIG. 4 is an enlarged explanatory view of a part a of the explanatory view (FIG. 2) by the cross section of the flow rate control and / or concentration means. Let d be the thickness of the flow control and / or concentration means 7.

Assuming that the wavelength of light in the air is λ ₁ , the refractive index is n ₁ = 1, the wavelength of light in the material resin of the flow control and / or concentration means is λ ₂ , and the refractive index is n ₂ , then λ ₁ = n ₂ λ ₂ ... I can be expressed. Since n ₁ <n ₂ , the light is reflected at the fixed end on the upper surface of the flow rate control and / or concentration means, so the phase of the reflected wave is reversed. Further, the flow rate control and / or concentration means lower surface and the solid support are added with a mixture of the reagent and the sample after addition of the mixture of the reagent and the sample, and the refractive index of light at this portion is determined by the material of the flow rate control means. Since it is smaller than the refractive index of the resin, light is reflected at the free end on the lower surface of the flow rate control and / or concentration means, and the phase of the reflected wave does not change. Accordingly, the conditions under which light is intensified are 2d = (m + 1/2) λ ₂ , (m = 0, 1, ₂ , 3...), Where d is the flow control and / or concentration means thickness. From λ ₁ = n ₂ λ ₂ (Formula I), 2n ₂ d = (m + 1/2) λ ₁ , (m = 0, ₁ , 2, 3...) is obtained. This is the condition for the strengthening of light. By selecting the refractive index n ₂ and thickness d of the resin so as not to meet this condition in the visible light wavelength range of about 400 nm to 800 nm, the reflected light and the flow rate control and / or the flow rate control and / or flow rate under the concentration means are selected. Alternatively, it is possible to avoid the development of a specific color without interference of light due to the reflected light on the upper surface of the concentration means, and the color tone of the non-test area can be confirmed accurately.

  Furthermore, when the flow control and / or concentration means of the present invention is molded from a transparent resin, an air layer is formed between the lower surface of the flow control and / or concentration means and the solid support after the addition of the reagent and the sample. It is desirable to adhere closely. FIG. 5 is an enlarged view of a part a of the explanatory view (FIG. 2) by the cross section of the flow rate control and / or concentration means. If the air layer 12 is formed between the lower surface of the flow rate control means and the solid support after the addition of the reagent and the sample, the color tone of the non-test area cannot be accurately confirmed. This is because the air layer prevents the light from the solid support corresponding to the position where the air layer exists, the non-test region, from reaching accurately, and the air layer exists on the lower surface of the flow control and / or concentration means as the cause. This is considered to be due to specular reflection due to the light, interference of light caused by the air layer, and a combination of these phenomena. In order not to form an air layer, the bottom surface of the flow rate control and / or concentration means is brought into close contact with the solid support included in the apparatus main body. For this purpose, when the flow control and / or concentration means is mounted on the apparatus main body, the bottom surface of the flow control and / or concentration means comes into full contact with the bottom surface of the solid phase support. It is desirable. For example, when the flow control and / or concentration means is mounted on the apparatus main body, the bottom surface of the flow control and / or concentration means is 0 to 0.5 mm from the plane on which the solid support is located when no pressure is applied from above. It is desirable to be located down the device. In this case, the bottom of the flow rate control and / or concentration means will hold down the solid support downward, but at the bottom of the solid support is an absorption site or spacer made of a relatively flexible material. Therefore, the solid support is not destroyed. Further, in order to make the bottom surface of the flow control and / or concentrating means come into close contact with the solid support included in the apparatus main body, the flow control and / or concentrating means is securely engaged or fitted with the apparatus main body. Need to be. For example, in the case of fitting, although not limited, three-point claw fitting or surface fitting may be adopted as a fitting method.

  Further, the reagent and sample addition part side part of the flow rate control and / or concentration means of the present invention does not include a part of the parabola in the cross-sectional shape, or if the part includes a part of the parabola, the focus is not in the test region. It is desirable. FIG. 6 is an explanatory diagram viewed from the top when the flow rate control and / or concentration means includes a part of a parabola and the focus is in the test region. When the light 11 is concentrated on the test area, there is a difference between the surroundings and the brightness, so accurate determination cannot be made.

  In addition, the flow control and / or concentration means of the present invention may be provided with a color for determining positive or negative or determining the degree of positive. FIG. 7 is an explanatory view of the apparatus provided with coloring for determining whether the flow rate control and / or concentration means is positive or negative or determining the degree of positive. The coloring may be a portion having an opening of the flow control and / or concentration means, for example, as long as it is provided on a surface visible from above when the means is attached to the apparatus main body. For example, the coloring is adjacent to the opening. It only has to be provided. The coloring 13 is for accurately performing the test result by preparing in advance a color (determination color) that serves as a positive determination threshold. Comparison with the determination color is performed, and positive / negative is determined based on the level of saturation of the color with the determination color and / or the lightness of the lightness. A plurality of determination colors may be provided. For example, the determination colors may be used to determine the degree of positiveness.

  In the detection method using the apparatus of the present invention, first, a target reagent for qualitative and quantitative analysis of an analyte is labeled with a specific binding reaction between the analyte and a ligand that specifically binds to the analyte. It is desirable to process in a state where it is easy to cause. The treatment method may be a chemical treatment method using various chemicals such as acids and bases, or may be either a physical treatment method using heating, stirring, ultrasonic waves, or both of them. good. Specifically, since the analyte is often present in a biological matrix such as a microorganism or cell in the sample, the sample is suspended in an acid solution, a base solution, a surfactant solution, a denaturant solution, or a buffer solution. The substance to be analyzed is extracted by disrupting the microorganism and cell matrix by stirring or heating. Next, the pH is adjusted so that the labeling reagent containing the ligand that specifically binds the suspension to the analyte and the analyte is likely to cause a specific binding reaction, or the concentration of inorganic salts is adjusted. Additives such as surfactants, polymer polymers, and basic compounds that enhance the reaction, and additives such as surfactants, polymer polymers, and basic compounds that reduce non-specific reactions are added. In the detection apparatus of the present invention, examples of the treatment liquid for treating the specimen include a specimen suspension composition described in JP-A-2003-279577. If the analyte in the sample is likely to cause a specific binding reaction with a labeling reagent containing a ligand that specifically binds to the analyte, the sample is suspended in a suspension that is susceptible to a specific binding reaction. It is possible to use only the specimen, or the specimen can be used as it is. When this series of sample processing is performed with a sample addition device (sample dropping container), the operation becomes simpler.

  The analyte to be analyzed in the method of the present invention is usually, but not limited to, an antigen or an antibody. The specimen is not limited, and examples include biological samples such as whole blood, serum, plasma, urine, saliva, sputum, sweat, and mucosal scrapings, and food extracts such as meat and plants. The ligand that binds to the analyte is typically an antibody that specifically binds to the antigen when the analyte is an antigen, and an antigen that specifically binds to the antibody when the analyte is an antibody. In addition, examples of the analyte-ligand combination include a receptor-ligand, a ligand-receptor, and the like. The labeling reagent is a conjugate in which the ligand and an appropriate labeling substance are bound, and as the labeling substance, a metal colloid such as a gold colloid, a nonmetal colloid such as a selenium colloid, a colored resin particle, a dye colloid, a colored liposome, etc. Insoluble particulate matter, alkaline phosphatase, peroxidase and other enzymes that catalyze a coloring reaction, fluorescent dyes, radioisotopes and the like.

  Next, the sample sample treated by the above-described method is brought into contact with the labeling reagent before being mixed on the solid phase support on which the capture reagent is immobilized, thereby mixing the labeling reagent-analyte complex. Let it form. In the apparatus of the present invention, for example, the flow rate control and / or concentration means capable of concentrating the reagent and the sample by controlling the flow rate is a container-like adapter for temporarily storing the specimen sample. In this case, the complex is contained in the adapter. It may be formed. There is an opening in the lower part of the adapter, and the specimen sample and the labeling reagent are supplied to the solid support through the opening while forming a complex. Contact between the labeling reagent and the specimen sample may be performed before adding to the adapter.

  When an analyte is contained in the sample, the sample and the labeling reagent are mixed to form a mixture by bringing the sample and the labeling reagent into contact with each other. The mixture of the specimen and the labeling reagent includes a mixture of the analyte and the labeling reagent, and further includes a complex of the analyte and the labeling reagent. The sample is treated by the above-described method so that the analyte and the ligand that specifically binds to the analyte are likely to cause a specific binding reaction, and the labeling reagent contacts an excess amount of the analyte. Therefore, many of the analytes efficiently form the complex only with the labeling reagent. Here, the capture reagent is a substance that specifically binds to the analyte, and the relationship between the capture reagent and the analyte is similar to the above-described relationship between the analyte and the labeling reagent. It can be an antigen, a receptor-ligand, a ligand-receptor, etc. The capture reagent and the labeling reagent may be the same substance, but when there is only one site that binds to the substance in the analyte, the labeling reagent-analyte-capture reagent complex is not formed. Therefore, in this case, the capture reagent and the labeling reagent need to bind to different sites of the analyte. The solid support may be any material as long as the sample specimen can be absorbed and flowed by capillary action. For example, the support is selected from the group consisting of nitrocellulose, cellulose acetate, nylon, polyethersulfone, polyvinyl alcohol, polyester, glass fiber, polyolefin, cellulose, and artificial polymers composed of mixed fibers thereof. The solid support is a film-like support having an arbitrary size, a capture reagent is immobilized on the membrane, and a capture reagent region is set on the membrane. The capture reagent may be immobilized on a solid support by a known method such as a method of adsorption, a method of chemically binding using a functional group such as an amino group, a carboxyl group, or an aldehyde group.

  Next, a sample containing the complex of the labeling reagent-analyte and the sample reagent is provided on a solid phase support on which a capture reagent that specifically binds to the analyte is immobilized, and the labeling reagent An analyte-capture reagent complex is formed. In the apparatus of the present invention, the specimen and the labeling reagent are added as a mixture to an adapter which is a flow control and / or concentration means, or mixed in the adapter to form a complex. The mixture containing the complex is fed onto the solid support through an opening at the bottom of the adapter. At this time, the supplied speed is controlled by the shape and size of the opening or by providing a filter between the opening below the adapter and the solid support. At this time, in the flow-through detection method, the complex of the labeling reagent and the analyte is captured by the capture reagent when passing through the solid support on which the capture reagent is immobilized, and the labeling reagent-analyte is analyzed. A substance-capture reagent complex is formed. The presence of the analyte can be determined by detecting the presence or absence of the labeling reagent captured on the solid support. Since the analyte and the labeling reagent are configured to contact with each other in advance at the site separated from the solid support, the analyte and the labeling reagent are sufficiently in contact with each other to form a complex.

  Therefore, the complex of labeled reagent-analyte-capture reagent can be easily and quickly formed simply by adding the sample to the solid support on which the capture reagent is immobilized. Detection (assay).

  The detection apparatus used in the method of the present invention may further contain a control reagent, and may further include a specimen addition part and an absorption part. The site containing the control reagent is called the control site and is included in the test area. The control reagent is not limited. For example, a substance to which a ligand in the labeling reagent binds can be used. In the flow-through detection method, the control reagent may be immobilized at a site different from the capture reagent on the membrane. The specimen addition part is a part for once absorbing the mixture of the specimen and the labeling reagent, and then supplying the absorbed mixture to the solid support on which the capture reagent is immobilized. The specimen adding part is preferably made of a porous material capable of absorbing a certain amount of liquid, and for example, a nonwoven fabric made of glass fiber or polystyrene may be used. The absorption site is a site having liquid absorbency that controls the flow of the sample by absorbing the sample that has passed through the capturing unit. In the flow-through detection method, for example, it may be provided below the membrane on which the capture reagent is immobilized. What is necessary is just to use a thing made from paper as an absorber pad, for example. Further, a spacer may be provided between the solid support and the absorption site. Here, the spacer needs to be liquid-permeable or liquid-absorbable so that the specimen sample reaches the absorption site, but there is no other limitation on the material and the function is not limited. For example, filter paper can be used, the rate at which the specimen sample is absorbed by the absorption site can be controlled by adjusting the liquid permeability and liquid absorbency, and the flow rate is controlled together with the flow rate control and / or concentration means. It can have a function. Further, if a thick material is used for the spacer, the solid support can be pressed against the bottom surface of the flow control and / or concentration means, and an air layer is formed between the lower surface of the flow control and / or concentration means and the solid support. Can be prevented.

  The present invention will be specifically described by the following examples, but the present invention is not limited to these examples. In the following Examples, for comparison, a commercially available Quick S-Influx A / B “Seiken” was used, and the adapter part newly molded was used as the apparatus of the present invention.

Example 1 Detection of influenza A virus using a flow-through detector (1) Preparation of colloidal gold antibody
Take 10 mL of gold colloid and adjust the pH to 7.0 with 100 mM potassium carbonate. Prepare anti-influenza A virus monoclonal antibody purified by dialysis, centrifugation and purification with 2 mM boric acid solution to a concentration of 100 μg / mL with 2 mM boric acid solution. An amount of 10 μg / mL final concentration of the prepared anti-influenza A virus monoclonal antibody is added to the colloidal gold with sufficient stirring. After 5 minutes, add 1 mL of 10% BSA and gently agitate with a rotator for 10 minutes. Transfer the entire volume to a centrifuge tube and centrifuge at 14000 rpm for 30 minutes at 4 ° C. After centrifugation, the supernatant is aspirated and discarded, and a solution containing 10 mM Tris-HCl buffer, 1% BSA, 150 mM sodium chloride in a sensitized version of colloidal gold colloid and anti-influenza A virus monoclonal antibody. Float with 1 mL.

(2) Drying of colloidal gold antibody The colloidal gold antibody produced in the previous section is sprayed onto a 10 mm x 300 mm non-woven fabric using a positive pressure spray device (BioDot, BioJet) at 8.0OD520, 10 μL / cm, and in an amount. . Subsequently, it was dried under reduced pressure in a vacuum apparatus for 1 hour to obtain a dry gold colloidal antibody pad. When used, it was cut at intervals of 7 mm.

(3) Immobilization of antibody on diagnostic membrane filter and preparation of detection device An example of immobilization of anti-influenza virus monoclonal antibody (mouse) on a nitrocellulose filter will be described.

  Prepare an anti-influenza virus monoclonal antibody (mouse) affinity-purified with a protein A column. The buffer in which the antibody is suspended is replaced with 0.1 mM trehalose-added 10 mM citrate buffer (pH 4.0) using a Sephadex G-25 gel filtration column.

  Dilute with 10 mM citrate buffer (pH 4.0) with 0.1% trehalose so that the absorbance at 280 nm is 1.0, and make an appropriate amount (for example, 10 μL / apparatus (device) for flow-through detection (diagnostic) equipment) D) Add dropwise onto the nitrocellulose attached to the detector, then leave it at 45 ° C for 40 minutes and dry.

(4) Detection method (in which a detection reagent is incorporated on the device side)
Do not block the test area and sample absorption area on the flow control and / or concentration means (adapter) that can control the flow rate of the detection device (device) and concentrate the reagent and sample in advance. In addition, it is incorporated so that it does not come off (FIG. 1B), and a sample of 500 μL, which is thought to contain influenza A virus, is filtered through a sample filtration filter and added dropwise. After the liquid is completely absorbed by the membrane member, if the membrane member where the anti-influenza virus monoclonal antibody is adsorbed is colored in the color of colloidal gold (eg, red to reddish brown), the influenza A virus in the sample Is confirmed to exist. If there is no change in color and the color of the membrane member remains, influenza A virus is not present in the sample.

  Flow control and / or concentration means capable of concentrating reagents and samples with controlled flow rate and / or concentration means molded with non-transparent colored resin, and flow control and / or concentration means synchronized with colorless transparent resin or solid support Tables 1 and 2 show the results of detection using the device of the present invention molded with a white or cream resin that is the color of the above.

The above data showed the average value of N = 2 measurements.
For a strongly positive sample, no significant time difference was observed between the conventional method and the method according to the present invention.

  When testing medium positive samples, the conventional method required more than 8 minutes to determine that the adapter was positive, but the improved adapter took 6-7 minutes and the transparent adapter took 5 minutes. It became possible to judge positive, and it was possible to shorten the time by up to 3 minutes or more and 4 minutes at the maximum.

  In addition to the above, by making the adapter transparent, external light can be taken in at a wider angle, and the merit of improving the visibility of the test area was also confirmed.

  In POCT (point of care) that requires rapid and simple, the total reaction (operation) time is often about 10 to 15 minutes, but in the present invention, the performance (sensitivity / specificity) is impaired. The time was shortened by more than 3 minutes (resulting in reducing the time to judgment to 70% of the conventional time).

  FIG. 8 shows a conventional apparatus (right side apparatus) in which the flow rate control and / or concentration means is formed of a blue non-transparent resin and the apparatus of the present invention in which the flow rate control and / or concentration means is formed of a colorless and transparent resin. FIG. 6 shows the color change of the initial reagent site with time when detection is performed. There is an opening in the lower part of the container-like adapter which is a flow control and / or concentration means so as to be the same as the capture reagent part on the nitrocellulose membrane. In the figure, there is a rhombus opening and a round opening at the bottom of one adapter, and this opening is matched in shape, size, and position to the capture reagent site on the nitrocellulose membrane. The diamond-shaped site is a test capture reagent site, and the round site is a control capture reagent site. 8A to 8C show the results after 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, and 8 minutes after addition of the sample, and the adapter was removed after 8 minutes. Indicates the state. As shown in the figure, in the conventional apparatus, after 6 to 8 minutes, although the capture reagent site is strongly colored, recognition of the color of the capture reagent site is inhibited by the color of the adapter. On the other hand, when the apparatus of the present invention is used, the control site can recognize the coloration after 1 minute. That is, immediately after the start of measurement, it can be confirmed that the detection has proceeded successfully. Moreover, coloring can be recognized in a test area | region after progress for 2 to 5 minutes. That is, if the detection is qualitative, it can be determined to be positive at this point.

  FIGS. 9 to 11 show a flow-through detection device used in this embodiment, and FIG. 12 shows a flow-through detection device in which the main body and the flow rate control and / or concentration means are integrated.

It is a figure which shows the flow through apparatus which has a flow control and / or concentration means. FIG. 1A shows the main body of the flow-through detection apparatus, FIG. 1B shows the flow rate control and / or concentration means (adapter) containing the labeling reagent, and FIG. 1C incorporates the labeling reagent into the sample addition device. A detection device is shown. Furthermore, FIGS. 4D and 4E show an overhead view of the flow control and / or concentration means containing the labeling reagent. It is a figure which shows the cross section of a flow control and / or concentration means, and is a figure which shows the path | route of the light which flows in between between an external structure and a flow control and / or concentration means. It is a figure which shows the cross section of a flow control and / or concentration means, and is a figure which shows the path | route of the light which reflects in an external structure and flows in. It is a figure which shows the bottom part of the cross section of a flow control and / or concentration means. It is a figure which shows the bottom part of the cross section of a flow control and / or concentration means, and is a figure which shows presence of an air layer. FIG. 5 is a top view of a light path when the flow control and / or concentration means includes a part of a parabola and the focus is on a test region. FIG. 5 shows a flow control and / or concentration means with a coloration for determining the degree of positive, negative or positive. It is a figure which shows the result of the detection using the conventional apparatus and the apparatus of this invention. It is a figure which shows the result of the detection using the conventional apparatus and the apparatus of this invention. It is a figure which shows the result of the detection using the conventional apparatus and the apparatus of this invention. It is a figure which shows the apparatus main body of the flow through type | mold detection apparatus which has a flow control and / or concentration means. It is a figure which shows the flow control and / or concentration means (adapter) of the flow-through type detection apparatus which has a flow control and / or concentration means. It is a figure which shows the detection apparatus which incorporated the flow control and / or concentration means (adapter) in the apparatus main body of the flow-through type detection apparatus which has flow control and / or concentration means. It is a figure which shows the flow through type | mold detection apparatus by which the flow control and / or concentration means were previously mounted | worn and integrated.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Detection apparatus 2 Solid phase support body 3 Spacer 4 Absorption site | part 5 Sample sample 6 Labeling reagent 7 Flow control and / or concentration means 8 Opening part (sample passage part)
9 capture reagent site 10 outer structure 11 light 12 air layer 13 coloring

Claims (5)

It consists of a resin body containing a solid phase support on which a capture reagent that specifically binds to the analyte in the sample is immobilized, and a flow rate control and / or concentration means, and specifically binds to the analyte. The solid phase support includes a test region in which a capture reagent is immobilized and a non-test region in which the capture reagent is not immobilized, and the flow rate control and / or concentration means is configured so that the reagent and the sample are in a solid phase. A flow-through detection device for performing an immunoassay capable of concentrating a reagent and a sample by controlling a flow rate moving to a support,
The flow control and / or concentration means is formed from a transparent resin in which the test region and the solid support including the test region and the solid support including the test region are visible through the flow control and / or concentration means. Well, with an aperture that fits in position,
Check the color in the test area and non-test area through the flow control and / or concentration means without touching the flow control and / or concentration means and without removing the flow control and / or concentration means from the detector. A flow-through detection device capable of confirming the coloration or non-coloration of a test region by comparison , wherein a labeling reagent is contained in a porous material, and the porous material is contained in a flow control and / or concentration means. Is a flow-through detection device .   The flow-through type detection apparatus according to claim 1, further comprising a sample addition device equipped with the porous material containing the labeling reagent, wherein the labeling reagent is contained in the porous material. The flow-through detection device according to claim 1 or 2, wherein the flow rate control and / or concentration means is integrated with the detection device. The flow-through detection device according to any one of claims 1 to 3 , wherein the flow rate control and / or concentration means is molded from polystyrene resin. The flow-through detection device according to any one of claims 1 to 4 , wherein the color of the outer structure on the side of the reagent and sample addition section of the flow rate control and / or concentration means is light.

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