JP4993757B2 - Immunochromatography equipment - Google Patents

Description

  The present invention relates to an immunochromatographic apparatus in which a plurality of strips for immunochromatography are housed in a housing.

  The immunochromatographic apparatus is an immunochromatographic method, that is, a sample is brought into contact with a sample development start position on an immunochromatographic development film made of a water-absorbing material, then developed in a predetermined direction by capillary action, and the development direction of the immunochromatographic development film It is an apparatus used in a method for analyzing a substance to be analyzed in a sample by observing a change in a detection zone (region where an antigen or antibody is immobilized) provided downstream.

  The immunochromatography method is roughly classified into a particle method (gold colloid method, latex method, etc.) and an enzyme method depending on the detection means. The particle method is inferior to the enzyme method in terms of the clarity of the staining line and the influence of specimen-derived coexisting substances (hemoglobin and bilirubin), but does not require an enzyme substrate, so it is necessary to add a developing solution after adding the sample. And the apparatus can be simplified. On the other hand, the enzyme method is excellent in the clarity of determination and not affected by the coexisting substance derived from the specimen, but requires an enzyme substrate, so that it is necessary to add a developing solution after addition of the sample separately.

FIG. 9 and FIG. 10 show a conventionally known immunochromatographic apparatus described in Patent Document 1 that can be used in the enzyme method.
9 and 10 includes a housing (housing upper lid 101a and housing substrate 101b), an immunochromatographic development film 109 made of an absorbent material and housed in the housing, and the immunochromatographic development film. The liquid developing material 107 (upstream side) and the liquid suction material 108 (downstream side) are included as means for determining the flow direction of the chromatographic development.
Further, the immunochromatograph apparatus 100 includes a sample supply means 102 for introducing a sample into the apparatus, a determination window 103 capable of observing the capture zone 195 on the immunochromatographic development film, an introduction section 104 as a development liquid supply means, And a developing solution storage chamber 111. The developing solution storage chamber 111 has a capacity capable of storing an optimal amount of developing solution, but can absorb an increment when an additional amount of liquid more than the optimal amount is supplied due to a supply error or the like. A liquid storage preliminary container 112 is provided.

  When a sample is added from the sample supply means 102 of the immunochromatography apparatus 100, the sample is supplied to the sample receiving region of the immunochromatographic development film 109. Subsequently, when the developing solution is added from the developing solution introduction unit 104 located upstream of the sample supply means 102, the developing solution storage chamber 111 is filled with the developing solution, and at the same time, the absorption of the developing solution by the liquid developing material 107 starts. Then, development on the immunochromatographic development film 109 is started. As a result of development on the immunochromatographic development film 109, it is possible to observe a color change in the capture zone 195 based on the presence or absence of the substance to be analyzed in the sample.

  As described above, in the immunochromatographic method using the enzyme method, since it is necessary to add a developing solution after adding a sample, it is common to have a developing solution supplying introduction unit in addition to the sample supplying introducing unit. It is. As far as the applicant knows, an enzyme immunochromatography apparatus in which a plurality of immunochromatographic strips are housed in a housing has not been commercially available so far, but for introduction of one developing solution for one immunochromatographic strip. When the part is provided, the addition of the developing solution is required several times, and the operation is expected to be complicated.

  In addition, due to the characteristics that immunochromatography equipment can be used for simple testing, it is expected to be used not only in well-equipped specialized laboratories, but also in homes, etc. It is also necessary to respond to use in environments that did not exist. For example, it is assumed that the stage on which the immunochromatograph device being deployed is not horizontal or that a light impact is accidentally applied. It is requested as a serious problem.

JP 2006-38600 A

  An object of the present invention is to provide an immunochromatography apparatus in which a plurality of strips for immunochromatography are housed in a housing, the operation can be simplified, and an accurate determination can be made even if a tilt occurs during deployment. Is to provide.

Said subject is according to the invention,
With two or more immunochromatographic strips in which a liquid spreading material, a spreading membrane having a sample receiving area and a capture area, and a liquid suction material are connected in this order along the spreading direction;
One developing solution storage chamber and one developing solution introduction part capable of supplying the developing solution to the developing solution storage chamber, and storing the immunochromatographic strips in the interior without any contact with each other. A possible housing;
An immunochromatographic apparatus comprising:
An end portion for absorbing a developing solution opposite to an end portion connected to the developing membrane in the liquid developing material is disposed in the developing solution storage chamber with respect to all the immunochromatographic strips;
This can be solved by an immunochromatograph apparatus characterized in that the developing solution distribution material that comes into contact with all of the developing solution absorbing end is disposed in the developing solution storage chamber.

  According to the present invention, by housing a plurality of strips for immunochromatography in a housing, the housing and the packaging unit price can be reduced. In addition, since only one developing solution storage chamber and one developing solution introduction part are provided for each of the plurality of immunochromatographic strips, it is possible to simplify the manufacturing process and simplify the operation of the user. Further, by setting the measurement items in one housing, it is possible to reduce mistakes in selecting the measurement items by the user and forgetting to put in the developing solution.

Hereinafter, the immunochromatography apparatus of the present invention will be described mainly with reference to the accompanying drawings.
The immunochromatography apparatus of the present invention is an improved invention of a conventionally known immunochromatography apparatus, includes a plurality of strips for immunochromatography, and has a specific shape of a developing solution introduction section, a developing solution storage chamber, and a developing device. It can have the same configuration as a conventionally known immunochromatography apparatus except that it includes a developing solution distribution means mainly composed of a liquid distribution material.

1 to 4 show an embodiment of the immunochromatographic apparatus of the present invention, which includes three immunochromatographic strips. 1 to 4 mainly includes a housing (housing upper lid 1a and housing substrate 1b) and three immunochromatographic strips 6 housed in the housing. .
Each immunochromatographic strip 6 includes an immunochromatographic developing film 9 made of an absorbent material, a liquid developing material 7 (upstream side) and a liquid as means for determining the flow direction of chromatographic developing on the immunochromatographic developing film. Includes suction material 8 (downstream).
The housing upper lid 1a has a sample supply means 2 for supplying a sample to the sample receiving zone 19 on the immunochromatographic development film, a judgment window 3 for observing the capture zone 20 on the immunochromatographic development film, and each immunochromatography. A developing solution introduction section 4 and a transpiration window 5 capable of supplying a developing solution are provided upstream of the graph test strip.

  The housing substrate 1b has a developing solution storage chamber 11 in which the developing solution supplied from the developing solution introduction section 4 can be stored, and can further have a liquid storage preliminary chamber 12 that can be provided as desired. . A developing solution distribution material 13 is disposed in the developing solution storage chamber 11, and a developing solution absorbing end portion 7 a that is an upstream end portion of the liquid developing material 7 is provided for each immunochromatographic strip 6. The developing solution storage chamber 11 is disposed in contact with the developing solution distribution material 13. 1 to 4, the bottom surface of the developing liquid storage chamber 11 and the entire area of the bottom surface of the developing liquid distribution material 13 are arranged so as to be in contact with each other. In the present invention, the end portion 7a for developing liquid absorption is disposed so as to be sandwiched between the bottom surface of the developing liquid storage chamber 11 and the lower surface of the developing liquid distribution material 13 in the present invention. You can also.

  The housing in the immunochromatographic apparatus of the present invention can have an appropriate shape and size according to the assay to be performed. The housing can also be fabricated from any material suitable for the assay being performed. The material used to fabricate the housing can be selected from materials that do not interfere with all reagents used in performing the assay, including the sample, sample medium, or components of the signal generating system. Preferably, the housing is made from a thermoplastic material or the like. Generally, the housing has a means (for example, a judgment window 3) for visually observing the capture zone 20 (s) on the surface of the immunochromatographic development membrane, from which the result of the assay can be measured. .

  The immunochromatography apparatus of the present invention is an immunochromatographic development film made of an absorbent material, which is housed in a housing, for capturing a member of a specific binding pair in a certain zone and moving the liquid from the zone by capillary action. Including. For example, in one aspect shown in FIGS. 1 to 4, the immunochromatographic developing film includes an absorbent material piece having one or more capture zones, that is, an absorbing immunochromatographic developing film 9. The immunochromatographic development film 9, the liquid suction material 8 and the liquid development material 7 are in a liquid receiving relationship, and the immunochromatographic development film 9 overlaps the liquid development material 7 on the upstream end side and the liquid suction material 8 on the downstream end side. (Regions 31, 41 respectively). By using the adhesive substrate 51 as needed, the connection between the liquid developing material 7, the immunochromatographic developing film 9, and the liquid suction material 8 can be strengthened. The material of the substrate is preferably made of paper or plastic, and the pressure-sensitive adhesive can be selected from those that do not interfere with all the reagents used for performing the assay. These are housed in the immunochromatography apparatus 10 and preferably can be confined in an immovable state in the immunochromatography apparatus 10. In the immunochromatography apparatus of the present invention, the number of immunochromatographic strips accommodated in the housing is not particularly limited as long as it is plural, but usually 2 to 10 strips can be accommodated.

The immunochromatography apparatus of the present invention is characterized by having a developing solution distribution means mainly composed of a developing solution introduction section having a specific shape, a developing solution storage chamber, and a developing solution distribution material. FIG. 5 shows an enlarged partial cross-sectional view of the structure of the developing liquid distributing means and the adjacent region thereof in one embodiment shown in FIGS.
The developing solution distribution means in the immunochromatography apparatus of the present invention is, for example, as shown in FIGS.
(A) One developing solution introduction part 4 having an upper opening 4a for adding a developing solution and a discharge port 4b provided at the bottom,
(B) one developing solution storage chamber 11 for storing the developing solution;
(C) In order to make the supply of the developing solution of the discharge port 4b and the liquid developing material 7 uniform and stable, the developing solution distribution material 13
Is provided. In the present invention, the developing liquid storage chamber needs not only to be able to store the developing liquid but also to be provided with a developing liquid distribution material. For example, the liquid storing chamber shown in FIG. 2 or FIG. The reserve chamber 12 does not correspond to the developing solution storage chamber in the present invention because the developing solution distribution material is not disposed.
The developing solution also includes a mixture to which various reagents including a buffer solution (for example, a signal generating component, a signal enhancing component, a nonspecific reaction suppressing component, etc.) are added.

  As shown in FIG. 5, in the immunochromatography apparatus of the present invention, the developing solution distribution material 13 is provided in the developing solution storage chamber 11, so Uniform and stable. The material of the developing liquid distribution material used in the present invention is not particularly limited as long as it is a hygroscopic material, but is a material having both liquid holding ability and liquid releasing ability, such as glass fiber, general filter paper , Sponge, cotton and the like. More specifically, for example, a fiber sheet made of glass fiber, polyvinyl alcohol (PVA) fiber, cellulose fiber, rayon fiber or the like, or a mixture of the fibers (for example, a combination of glass fiber and PVA fiber, cellulose and glass) A fiber sheet made of a combination of glass fiber and PVA fiber, a fiber sheet made of a combination of cellulose and glass, and a fiber sheet made of rayon.

  In the embodiment shown in FIGS. 1 to 6, the developing solution amount is supplied with the optimum amount for performing the test, and the developing solution storage chamber 11 has a capacity capable of storing the optimum amount. The development liquid storage chamber 11 and the liquid storage preliminary chamber 12 are connected through the opening 55 (a plurality can be provided) so that the increment when the development liquid amount exceeding the optimum amount is supplied due to a supply error or the like flows into the liquid storage preliminary chamber 12. ing.

Further, as shown in FIGS. 1 and 5, a developing solution introduction section 4 that can be used for supplying the developing solution to the liquid developing material 7 or the developing solution distributing material 13 can be included. The developing solution introduction part 4 (the shape seen from above) is not particularly limited, but may be, for example, a cylinder, a cone, a prism, or a pyramid as shown in FIG. 7 or FIG. A pyramid shape is preferable.
The shape (shape seen from the side surface) of the developing solution introduction part in the present invention is not particularly limited, but for example, a pyramid shape (for example, a quadrangular pyramid, a triangular pyramid, a polygonal pyramid), a hemispherical shape, a conical shape, A cylindrical shape or a prismatic shape (for example, a quadrangular prism, a triangular prism, or a polygonal prism) can be used, and a pyramid shape is particularly preferable in terms of easy discrimination and supply of the developing liquid introduction portion.

The position and size of the upper opening of the developing solution introduction unit can be determined as appropriate according to the shape and the increase / decrease of the size of the opening accompanying the increase / decrease of the number of test strips. For example, when three test strips having a width of 6 mm are stored, the opening diameter thereof is, for example, 0.5 cm or more × 0.5 cm or more, preferably 0.5. It can be ˜2 cm × 0.5-5 cm, more preferably 0.5-1 cm × 0.5-3 cm.
Further, the shape, position and size of the bottom discharge port can be determined so as to coincide with the increase / decrease in the number of test strips and the shape, position and size of the next developing liquid flow path as in the upper opening.

  EXAMPLES Hereinafter, the present invention will be specifically described by way of examples, but these do not limit the scope of the present invention.

Example 1
(1) Production of strip for enzyme method immunochromatography The immunochromatographic test strip shown in FIG. 3 was produced by the following method.

(1-1) Preparation of mite allergen-immobilized membrane 9 Mite allergen (manufactured by Greer) is pasted on a nitrocellulose membrane (HF240: manufactured by Nippon Millipore, length 57 mm x width 40 cm), and a liquid suction material 8 in the vertical direction is attached. The film was applied linearly at a location of about 15 mm from the end to be attached (downstream side) under the condition of 1 μL / cm.
As it was, after standing and drying at room temperature for 1 hour, it was stored in a silica gel desiccator and dried overnight at room temperature to obtain a mite allergen-immobilized membrane.

(1-2) Production of Absorption Pad (Liquid Suction Material 8) A glass pad (A / B: manufactured by Nippon Pole Co., Ltd.) was cut into a size of 30 mm in length and 40 cm in width to obtain an absorption pad.

(1-3) Production of Development Pad (Liquid Development Material 7) A glass pad (AP25: manufactured by Nihon Millipore) was cut into a size of 20 mm in length and 40 cm in width to obtain 5-bromo-4chloro-3-indolyl phosphate ( A 5-bromo-4-chloro-3-indolyl phosphate (BCIP) substrate solution (manufactured by Sigma) was added at a concentration of 2 μL / min. It was applied linearly under the condition of cm. As it was, it was allowed to stand at room temperature for 1 hour and dried, then stored in a silica gel desiccator, dried overnight at room temperature, and used as a development pad.

(1-4) Preparation of immunochromatographic test strip An adhesive sheet (manufactured by Nihon Millipore) is cut into a size of 84 mm in length and 40 cm in width. An absorbent pad and a development pad were attached to each other with an overlap of about 8 mm in the direction. Subsequently, it cut | disconnected by width 4mm width, and was set as the immunochromatography test strip.

(2) Production of reaction cassette (immunochromatography apparatus 10) The immunochromatography apparatus 10 shown in FIGS. 1 to 4 was produced by the following method.

(2-1) Production of developing solution supply pad (developing solution supply hygroscopic material 13) Glass-PVA (Type A / E: manufactured by Nippon Pole), cellulose-glass (LF1: manufactured by Whatman), rayon (8-S) : Made by Schleicher & Schuell), each was cut to a size of 28 mm × 11 mm, and used as a developing solution supply pad. All thicknesses are 0.3 mm.

(2-2) Production of Reaction Cassette (Immunochromatograph Device 10) Three test strips were accommodated in a housing capable of accommodating three test strips, and further produced on the bottom side of the developing solution storage container 11 as described above. One developing solution supply pad was accommodated, and the ends of the three test strips were placed in contact with the developing solution supply pad. As a comparative example, a reaction cassette that does not contain a developing solution supply pad was prepared.

(3) Preparation of enzyme-labeled antibody reaction solution Alkaline phosphatase-labeled anti-human IgE antibody solution (manufactured by DPC) in 2% Triton X-100, 0.15 mol / L NaCl, 0.1 mol / L HEPES buffer (pH 8) solution ) Was added to prepare an enzyme-labeled antibody reaction solution.

(4) Preparation of developing solution A 0.1 mol / L CHES buffer solution (pH 10) and a 5 mmol / L magnesium chloride solution were prepared.

(5) Measurement of human serum using a reaction cassette Using the reaction cassette prepared above, 30 μL of human serum [positive sample (2+) or negative sample (−)] and 100 μL of enzyme-labeled antibody reaction solution are mixed. Then, 30 μL of each of the mixed solutions is added to each of the round holes (total of three locations) of the reagent supply means 2 of the reaction cassette, and then the developing solution is immediately added to the rectangular developing solution introducing unit 4 according to the following conditions. It added to the location, it was made to react at room temperature in a stationary state, and the linear coloring degree which appears in the judgment window 3 after 40 minutes was judged. When an antibody that reacts with mite allergen is contained in human serum, blue color is developed. The strongest color degree was 3+, the next medium color degree was 2+, the weak color degree was +, and no color was observed. Moreover, the thing which color development was not recognized on a line (what was distorted), the raise of a background, generation | occurrence | production of nonspecific color development, etc. were recognized as the error. A positive specimen is determined if color development of + or higher is shown.
Human serum used was determined to be a positive specimen [Class 2 (2+)] or a negative specimen [Class 0 (−)] by the Unicap-specific IgE method (manufactured by Fadia).

(5-1) Influence of inclination of immunochromatograph apparatus (housing) and influence of material of developing solution supply pad (1) The sample and developing solution were added in a horizontal state, and then allowed to stand with an inclination of 15 degrees. In the case (horizontal → tilt), (2) When the sample and the developing solution are added in a state where there is a 15 degree inclination, and left to stand with the inclination (inclination → inclination), (3) there is a 15 degree inclination When the sample and the developing solution are added in a state and then left to stand in a horizontal state (tilt → horizontal), (4) When the sample and the developing solution are added in a horizontal state and left to stand as it is (horizontal → horizontal) In these four conditions, the degree of color development was examined under the condition that there were no three types of developing solution supply pads (glass-PVA, cellulose-glass, rayon) and no developing solution supply pad. In addition, the inclination achieved the predetermined inclination angle by maintaining the right side of the housing in a state where the left side of the housing is supported as a fulcrum in the deployment direction.
The amount of the developing solution used for the examination is the maximum amount of the developing solution in the developing solution supply pad to be used, and the amount of the developing solution in one immunochromatographic test strip is 200 μL. A total amount of 600 μL was used. Specifically, 800 μL for glass-PVA (maximum holding amount = 200 μL), 680 μL for cellulose-glass (maximum holding amount = 80 μL), 740 μL for rayon (maximum holding amount = 140 μL), no pad In this case, 600 μL was used. Further, the rightmost point (that is, the highest point) from the inclined center point was examined as the position below the developed droplet. In addition, three experiments were repeated.
The results are shown in Table 1 [positive specimen (2+)] and Table 2 [negative specimen (−)].

  In the positive specimen (2+), when glass-PVA was used, 2+ color development was observed under all conditions. When cellulose-glass or rayon was used, 2+ color development was observed in the left and center immunochromatographic test strips, whereas in the right one, color development was weakened. On the other hand, when the pad was not used, when the sample and the developing solution were added in the horizontal state of the condition (4) and allowed to stand as it was, 2+ color development was observed, but the conditions (1) to (3) When placed in the tilted state, color development was weakened and many errors were observed.

  In the negative specimen (-), when glass-PVA, cellulose-glass, or rayon was used, no color development was observed. When the pad was not used, color development was not recognized under the condition (1) or (4), but an error was sometimes observed under the condition (2) or (3).

  From these results, it became clear that by using the developing solution supply pad, the specimen can be accurately evaluated without being affected by the tilt of the immunochromatograph apparatus and the position placed in the immunochromatograph apparatus.

  The immunochromatography apparatus of the present invention can be applied to analysis applications using antigen-antibody reaction.

It is a top view of the housing upper cover in one mode of the immunochromatography device of the present invention. In the one aspect | mode of the immunochromatography apparatus of this invention shown in FIG. 1, it is a top view which shows the state which removed the housing upper cover. It is a perspective view of the strip for immunochromatographs in the one aspect | mode of the immunochromatograph apparatus of this invention shown in FIG. FIG. 2 is a rear view of a housing upper lid in one embodiment of the immunochromatography apparatus of the present invention shown in FIG. 1. FIG. 2 is an enlarged partial cross-sectional view showing the structure of a developing solution distribution means and its adjacent region in one embodiment of the immunochromatography apparatus of the present invention shown in FIG. It is a top view of the housing substrate in the one aspect | mode of the immunochromatography apparatus of this invention shown in FIG. It is explanatory drawing which shows typically the various side surface shapes of the developing solution introduction part in the immunochromatography apparatus of this invention. It is explanatory drawing which shows typically the various upper surface shapes of the developing solution introduction part in the immunochromatography apparatus of this invention. It is a perspective view of the conventionally well-known immunochromatograph apparatus which accommodated one strip for immunochromatographs. FIG. 10 is a perspective view showing the conventionally known immunochromatography apparatus shown in FIG. 9 in a state where it is disassembled into a housing top cover, an immunochromatographic strip, and a housing substrate.

Explanation of symbols

10: Immunochromatography apparatus;
DESCRIPTION OF SYMBOLS 1 ... Housing; 1a ... Housing upper cover; 1b ... Housing board | substrate;
2 ... Sample supply means; 3 ... Judgment window; 4 ... Developing liquid introduction part;
5 ... evaporation window; 6 ... strip for immunochromatography;
7 ... Liquid spreading material; 8 ... Liquid suction material;
9: Immunochromatographic development membrane;
DESCRIPTION OF SYMBOLS 11 ... Development liquid storage chamber; 12 ... Liquid storage preliminary chamber; 13 ... Development liquid distribution material;
19 ... Sample receiving zone; 20 ... Capture zone.

Claims (1)

With two or more immunochromatographic strips in which a liquid spreading material, a spreading membrane having a sample receiving area and a capture area, and a liquid suction material are connected in this order along the spreading direction;
One developing solution storage chamber and one developing solution introduction part capable of supplying the developing solution to the developing solution storage chamber, and storing the immunochromatographic strips in the interior without any contact with each other. A possible housing;
An immunochromatographic apparatus comprising:
An end portion for absorbing a developing solution opposite to an end portion connected to the developing membrane in the liquid developing material is disposed in the developing solution storage chamber with respect to all the immunochromatographic strips;
An immunochromatograph apparatus, wherein a developing solution distribution material that contacts all of the developing solution absorbing ends is disposed in the developing solution storage chamber.

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