JP6254994B2 - Inspection kit - Google Patents

Description

  The present invention relates to a test kit.

  Conventionally, as an immunoassay, immunity in which the labeled antibody is held releasably in a wet state and the presence or absence of the antigen is determined using an immunochromatographic test strip on which the antigen capturing antibody and the labeled capturing antibody are immobilized. Measurement methods are known. Such a test strip is not housed in a case body made of plastic or the like, and the end of the strip-shaped test strip is directly put into the sample extract, so that the reaction proceeds on the test strip. This is a horizontal cassette type that is installed in a case main body made of plastic and the like, and the specimen extract is dropped onto the test strip inside the case main body from the specimen dropping opening so that the reaction proceeds on the test strip. There are two types of test strips.

  In the dipstick type test strip, the test strip is inserted in a state in which the test strip is set up in the sample extract in the container, so that the development direction of the sample extract is opposite to the heavy load. On the other hand, in the case of cassette-type test strips installed horizontally in the case body, the case body is placed horizontally on a flat surface such as a desk and used, so the direction in which the specimen extract dropped onto the specimen dropping entrance is developed Becomes a horizontal direction perpendicular to the direction of gravity (see, for example, non-patent literature). In this regard, they differ greatly in the direction of liquid deployment on the test strip against gravity.

  Here, FIG. 34 shows an inspection kit 100 in which a cassette type inspection strip 110 that is installed horizontally is installed on the case body 101. In fact, this type of inspection kit 100 has a case body 101 in which a lid 102 made of a resin material such as plastic is fitted and fixed to a base 103 made of a resin material such as plastic. The test strip 110 is installed horizontally in the case body 101.

  The lid 102 has a sample dropping opening 105 and a determination opening 106 formed on the upper surface, and the sample pad SP of the test strip 110 installed on the base 103 is exposed from the sample dropping opening 105 to the outside. At the same time, the membrane M of the test strip 110 can be exposed from the determination opening 106 to the outside. Thus, in the test kit 100, the sample extract L can be dropped from the sample dropping opening 105 to the sample pad SP of the test strip 110 in the case body 101, and thus whether or not a line has developed on the membrane M of the test strip 110. The user can visually recognize this through the determination opening 106 and can determine the presence or absence of the antigen in the sample extract based on the presence or absence of the expression of the line.

  In practice, as shown in FIG. 35A, the test strip 110 is formed on a strip-like soft support material 111 along the deployment direction x, the sample pad SP, the conjugate pad CP, the membrane M, and the absorbent pad AbP. Are arranged in order, and when the specimen extract is dropped on the sample pad SP, the specimen extract can be developed in the order of the conjugate pad CP, the membrane M, and the absorption pad AbP by capillary action.

  In this case, the conjugate pad CP includes, for example, a labeled antibody A1 that specifically reacts with a predetermined antigen, and another antigen that specifically reacts with a different type of antigen from the antigen that the labeled antibody A1 specifically reacts with. The labeled antibody B1 is held releasably in a wet state. On the other hand, in the membrane M, one test line TL1, another test line TL2, and a control line CL are sequentially arranged from the conjugate pad CP to the absorption pad AbP, and the sample that has passed through the conjugate pad CP. The extract can pass through one test line TL1, another test line TL2, and the control line CL in this order by capillary action.

  In this case, one antigen capture antibody A2 that specifically reacts with an antigen that specifically reacts with one labeled antibody A1 of the conjugate pad CP is immobilized on one test line TL1 of the membrane M. . In addition, another antigen-capture antibody B2 that reacts specifically with another antigen that specifically reacts with another labeled antibody B1 of the conjugate pad CP is immobilized on another test line TL2 of the membrane M. Yes. Furthermore, a labeled capture antibody r that specifically reacts with all the labeled antibodies A1 and B1 of the conjugate pad CP is immobilized on the control line CL of the membrane M.

  When using the test kit 100 in which the test strip 110 having such a configuration is installed horizontally, the user collects the specimen by rubbing the subject's pharynx and nasal cavity with a cotton swab tip cotton. The tip cotton of the cotton swab is immersed in a chemical solution in the extraction tube, and the chemical solution in the extraction tube is stirred with a cotton swab to obtain a sample extract in which the collected sample is released into the chemical solution.

  Next, this sample extract is dropped into the sample dropping opening 105 of the test kit 100 installed horizontally on a flat surface so that the sample extract is absorbed by the sample pad SP. At this time, as shown in FIG. 35B in which the same reference numerals are assigned to the corresponding parts to FIG. 35A, the sample pad contains an antigen G that specifically reacts with one labeled antibody A1. When the specimen extract in the SP is spread on the conjugate pad CP by capillary action, one labeled antibody A1 reacts specifically with the antigen in the specimen extract to produce complex C, and the complex C Deploy to membrane M along deployment direction x.

  In the membrane M, the antigen capture antibody A2 and the antigen G of the complex C are combined on the test line TL1 to which the antigen capture antibody A2 that specifically reacts with the antigen G is immobilized. And the complex C is captured by the antigen-capturing antibody A2, and the line can be expressed on the test line TL1 by the labeled antibody A1 of the captured complex C. In the other test line TL2 to which the antigen capture antibody B2 that does not cause a specific reaction with the antigen G is immobilized, the antigen G is not captured by the antigen capture antibody B2 and the line is not expressed.

  Thus, the user recognizes that the line has developed from the determination opening 106 of the test kit 100 to the test line TL1 in the membrane M by visual observation or a fluorescent reader, so that the predetermined antigen G is extracted from the specimen extract. It can be determined that it is contained within. In the test strip 110, the specimen extract that has passed through the test lines TL1 and TL2 of the membrane M can also be developed on the control line CL and absorbed as it is into the absorption pad AbP. At this time, in the control line CL of the membrane M, each of the labeled antibodies A1 and B1 of the conjugate pad CP dissolved in the sample extract specifically reacts with the labeled capture antibody r, and the labeled capture antibody r is labeled with the labeled antibody. A1 and B1 are captured, and a line indicating that is expressed in the control line CL. As a result, the user can visually confirm to what extent the specimen extract containing the labeled antibodies A1 and B1 of the conjugate pad CP is developed on all the test lines TL1 and TL2 on the membrane M.

Nichirei Biosciences Co., Ltd. “In-vitro diagnostic pharmaceutical influenza virus kit Immunofine ™ FLU” [online], searched on November 7, 2014, Internet (URL: http://www.nichirei.co.jp/bio/ products / pdf2 / 522061_2.pdf)

  However, when using the test kit 100 having such a configuration, a sample is collected from the subject's nasal cavity, pharynx, etc. at the medical site, and then a sample extract containing the sample is prepared, and the sample extraction is further performed. A prescribed amount of liquid is dropped on the sample pad SP on the spot to determine whether the line appears on the test lines TL1 and TL2 of the membrane M. Therefore, in such a test kit 100, a clear line that can be identified by the user is expressed in the test lines TL1 and TL2 in a short time, and it is possible to quickly determine whether or not the antigen G is present in the sample extract. It is desirable to be able to do it in a timely and reliable manner. For this purpose, it is necessary to capture more complex C in a short time and reliably with the antigen capture antibodies A2 and B2 immobilized on the membrane M.

  In addition, when using the conventional horizontal cassette type test kit 100, the sample extract exceeds the preset amount (for example, about 2 drops (about 55 μL)) and drops on the sample pad SP. In other words, the phenomenon that the line that should be expressed in either of the test lines TL1 or TL2 of the membrane M is difficult to express clearly, or that the expression of the line itself is delayed occurs. There is a problem that it is difficult to determine whether or not antigen G is contained in a short time.

  On the other hand, when a dipstick type test strip is used, since the sample absorbing portion at the end of the test strip is immersed in the sample extract, the cassette type described above can be used even if the amount of the sample extract is large. The problem as in the test strip 110 of FIG.

  Considering the difference between the cassette-type test strip 110 used in the horizontal position and the dipstick-type test strip used vertically, the dipstick-type test strip used in the vertical position Since all of the sample extract absorbed in the sample absorption part rises against the gravity in the test strip due to capillary action, any other sample extract cannot touch the surface of the conjugate pad. I can say no. On the other hand, in a cassette-type test strip that is used horizontally, if a sample extract exceeding the specified amount is applied, the sample extract exceeding the specified amount is transferred from the sample pad SP to the conjugate pad by capillary action. It is presumed that it has flowed onto the surface of the conjugate pad CP before deploying to CP.

  In this way, it can be estimated that there is a difference in at least how the specimen extract is developed between the cassette-type test strip 110 used in the horizontal orientation and the dipstick-type test strip used in the vertical position, It can also be inferred that the specimen extract may have some adverse effect on the expression of the test line by flowing out onto the surface of the conjugate pad.

  Although it is difficult to accurately explain what mechanism causes the occurrence of the above-described problems, the conventional test kit 100 using the test strip 110 used in the horizontal position is used by the user. When a sample extract exceeding the specified amount is dropped on the sample pad SP by mistake, the line is not clearly expressed on the test lines TL1 and TL2, and it is difficult to determine the presence or absence of antigen G in a short time It is true that

  Therefore, the present invention has been made in consideration of the above points, and whether or not a sample is present in the sample extract even when the user drops a sample extract of a specified amount or more on the sample pad, An object is to propose a test kit that can be easily determined in a shorter time than in the past.

  In order to solve this problem, the test kit according to claim 1 is a test kit in which a test strip is installed in a case body, and the test strip includes a sample pad to which a sample extract containing a sample is applied. A labeling substance that specifically reacts with the analyte developed from the sample pad by capillary action to form a complex, and the conjugate pad that is releasably retained in a wet state; A sample having a complex and a membrane having a detection region on which a capture substance specifically reacting is immobilized, and the sample body of the test strip is exposed to the outside on the upper surface of the case body A drip opening and a determination opening that exposes the membrane of the test strip to the outside are formed. A cover member is provided between the first pad and the detection region of the membrane so as to cover at least the conjugate pad.

  According to the test kit of the present invention, even when a sample extract of a specified amount or more is applied to the sample pad by the user, the sample extract is guided into the conjugate pad by the cover member, and the labeling substance and the sample extract It is easy to obtain a complex by specifically reacting with the sample, and as a result, a larger amount of the complex can be captured by the membrane capture substance and the time required for the capture substance to capture the complex is shortened. Thus, it can be easily determined in a shorter time than before whether or not a sample is present in the sample extract.

It is the schematic which shows the structure of the test | inspection kit of this invention. 2A is a schematic diagram illustrating a configuration of a test strip used in the test kit according to the first embodiment, and FIG. 2B is an exploded view illustrating a configuration when the test strip of FIG. 2A is disassembled. FIG. 3A is a schematic diagram illustrating a configuration of a test strip used in the test kit according to the second embodiment, and FIG. 3B is an exploded view illustrating a configuration when the test strip of FIG. 3A is disassembled. 4A is a schematic diagram showing a configuration of a test strip with a 5 mm first cover tape attached thereto, and FIG. 4B is a schematic diagram showing a configuration of the test strip with a 5 mm second cover tape attached thereon. FIG. FIG. 4 is a schematic diagram showing the configuration of a test strip with a 9 mm cover tape attached, and FIG. 4D is a schematic diagram showing the configuration of a test strip with a 12 mm cover tape attached. It is the schematic which compared each area | region where the 5mm 1st cover tape, 5mm 2nd cover tape, 9mm cover tape, 12mm cover tape, and the cover tape which covers the whole membrane surface are affixed. It is the table | surface which put together the line intensity | strength, the detection time of each line, and special remarks about the conventional test strip using an adenoplate and without a cover tape. It is the table | surface which put together the line intensity | strength, the detection time of each line, and special remarks about the test strip which used the adenoplate and provided the full surface cover tape. It is the table | surface which put together the line intensity | strength, the detection time of each line, and special remarks about the test strip which provided the 5mm 1st cover tape using an adenoplate. It is the table | surface which put together the line intensity | strength, the detection time of each line, and special remarks about the test strip which provided the 2mm cover tape using an adenoplate. It is the table | surface which put together the line intensity | strength, the detection time of each line, and special remarks about the test strip which provided the 9mm cover tape using an adenoplate. It is the table | surface which put together the line intensity | strength, the detection time of each line, and special remarks about the test strip which provided the 12mm cover tape using an adenoplate. It is the graph which showed the relationship (1) between sample amount and line intensity. It is the graph which showed the relationship (1) of the sample amount and T line detection time. It is the graph which showed the relationship (2) between sample amount and line intensity. It is the graph which showed the relationship (2) of the sample amount and T line detection time. It is the table | surface which put together the line intensity | strength, the detection time of each line, and special remarks about the conventional test strip without a cover tape using a strep A plate. It is the table | surface which put together the line intensity | strength, the detection time of each line, and special remarks about the test strip which provided the full surface cover tape using the strep A plate. It is the table | surface which put together the line intensity | strength, the detection time of each line, and special remarks about the test strip which provided the 5mm 1st cover tape using the strep A plate. It is the table | surface which put together the line strength, the detection time of each line, and special notes about the test strip which provided the 5mm 2nd cover tape using the strep A plate. It is the table | surface which put together the line intensity | strength, the detection time of each line, and special notes about the test strip which provided the 9mm cover tape using the strep A plate. It is the table | surface which put together the line strength, the detection time of each line, and special notes about the test strip which provided the 12mm cover tape using the strep A plate. It is a table (1) that summarizes line strength, detection time of each line, and special notes for a conventional test strip without cover tape using an FLU plate. Table 2 summarizes line strength, detection time for each line, and special notes for a conventional test strip that uses FLU plates and has no cover tape. Table (1) that summarizes line strength, detection time of each line, and special notes for a test strip using a FLU plate and provided with a full cover tape. Table 2 summarizes line strength, detection time of each line, and special notes for a test strip using a FLU plate and provided with a full cover tape. FIG. 5 is a table (1) that summarizes line strength, detection time of each line, and special notes for a test strip that uses a FLU plate and is provided with a 5 mm first cover tape. Table 2 is a table (2) that summarizes line strength, detection time of each line, and special notes for a test strip provided with a 5 mm first cover tape using an FLU plate. Table (1) that summarizes line strength, detection time of each line, and special notes for a test strip using an FLU plate and provided with a 5 mm second cover tape. It is a table (2) that summarizes the line strength, detection time of each line, and special notes for the test strip using the FLU plate and provided with the 5 mm second cover tape. Table (1) that summarizes line strength, detection time of each line, and special notes for a test strip using an FLU plate and provided with a 9 mm cover tape. Table (2) that summarizes line strength, detection time of each line, and special notes for a test strip using an FLU plate and provided with a 9 mm cover tape. It is the table | surface (1) which put together the line intensity | strength, the detection time of each line, and special notes about the test strip which provided the 12mm cover tape using the FLU plate. Table 2 is a table (2) that summarizes line strength, detection time of each line, and special notes for a test strip using an FLU plate and provided with a 12 mm cover tape. It is the schematic which shows the structure of the conventional test | inspection kit. FIG. 35A is a schematic diagram showing a configuration of a test strip used in a conventional test kit, and FIG. 35B is a schematic diagram showing a state in which a specimen extract containing an antigen develops the test strip.

Hereinafter, modes for carrying out the present invention will be described. The description will be in the following order.
1. 1. First embodiment 1-1. Configuration of test kit and test strip 1-2. Actions and effects Second Embodiment 3. FIG. Other Embodiment 4 Verification test 4-1. Test strip using blue colored latex particles 4-2. Test strip using colloidal gold particles 4-3. About test strips used to detect influenza virus

(1) First Embodiment (1-1) Configuration of Test Kit and Test Strip In FIG. 1, in which parts corresponding to those in FIG. 34 are assigned the same reference numerals, 1 indicates a test kit according to the present invention, 34 differs from the conventional test kit 100 shown in FIG. 34 in the configuration of the test strip 2 installed in the case body 101, and is characterized in that the conjugate pad CP is covered with the cover tape 5.

  In practice, the test strip 2 has a sample pad SP, a conjugate pad CP, a membrane M, and an absorption pad AbP arranged in this order, as in the prior art, and the specimen extract that has been absorbed by the sample pad SP. The conjugate pad CP, the membrane M, and the absorbent pad AbP can be developed in this order along the deployment direction x by capillary action. Further, the test strip 2 is fixed to the base 103 by being fitted into a region surrounded by a plurality of holding projections 115 formed in the base 103, and the sample pad SP and the conjugate pad CP are fixed. The membrane M and the absorbent pad AbP can be positioned at predetermined positions in the base 103.

  In the base 103, with the test strip 2 fixed, corresponding protrusions (not shown) on the inner wall surface of the lid body 102 are fitted into the plurality of fitting portions 116 on the inner wall surface, respectively. Can be mounted. Thereby, the base 103 and the lid 102 can be integrated to form the case main body 101. The test strip 2 installed in the case body 101 is exposed to a part of the sample pad SP from the specimen dropping opening 105 of the lid 102 and from the determination opening 106 of the lid 102 to the membrane M. The test lines TL1 and TL2 and the control line CL can be exposed to the outside.

  On the other hand, in the test strip 2, the cover tape 5 covering the entire surface of the conjugate pad CP from a partial region of the sample pad SP, the conjugate pad CP, and the absorbent pad AbP are covered by the lid 102. , It can be unexposed to the outside. As described above, the test strip 2 is affixed so as to avoid the region where the sample dropping opening 105 and the determination opening 106 are arranged, and the cover tape 5 is not exposed to the outside from the lid 102. Therefore, the lid 102 makes it difficult for dust and the like in the atmosphere to adhere to the periphery of the cover tape 5, and can prevent the cover tape 5 from peeling off from the conjugate pad CP. It is made like that.

  Next, the configuration of the test strip 2 used in the test kit of the present invention will be described in detail. As shown in FIGS. 2A and 2B, the test strip 2 is arranged on the support 111 in the order of the sample pad SP, the conjugate pad CP, the membrane M, and the absorbent pad AbP along the deployment direction x, of which The structure covers the entire surface of the conjugate pad CP from the partial region of the sample pad SP with the cover tape 5.

  In the case of this embodiment, the sample pad SP is laminated on the conjugate pad CP between the sample pad SP and the conjugate pad CP, and the sample pad SP rises by the thickness of the conjugate pad CP. The sample pad side laminated portion 8a is formed. The conjugate pad CP absorbs the sample extract absorbed in the sample pad SP by the sample pad side laminated portion 8a by capillary action, and extracts the sample into a plurality of kinds of immobilized labeled antibodies (not shown). Antigens in the fluid can react specifically.

  The cover tape 5 is formed of a non-permeable synthetic resin member such as a plastic film or a polyester film, for example, and the sample extract dropped onto the sample pad SP does not permeate from the sample pad SP to the conjugate pad CP. The specimen extract can be led to In the case of this embodiment, the cover tape 5 is formed in a strip shape, the width is selected to be the same as the width of the sample pad SP and the conjugate pad CP, and the region covered from one end to the other end of the cover tape 5 In this case, the sample pad SP and the conjugate pad CP can be made unexposed to the outside.

  In the case of this embodiment, the cover tape 5 is the sample pad SP that is covered with the lid 102 and not exposed to the outside from the specimen dropping opening 105 when the test strip 2 is installed in the case body 101. It is formed so that one end can be arranged in the non-exposed region portion. As a result, the test strip 2 can be in a state in which the sample pad SP is exposed in the entire region of the specimen dropping opening 105, so that when the specimen extract is dropped from the specimen dropping opening 105 to the sample pad SP, the sample pad SP The specimen extract can be reliably dropped directly onto the surface of the sample pad SP without being hindered by the cover tape 5 from dropping the specimen extract onto the surface of the sample pad SP.

  In the case of this embodiment, the cover tape 5 has the other end disposed at the end of the conjugate pad CP located at the boundary between the membrane M and the conjugate pad CP, and the non-exposed region of the sample pad SP. The entire surface of the conjugate pad CP can be covered and the surface of the membrane M can be exposed to the outside without covering the surface of the membrane M. As a result, in the test strip 2, the test lines TL1 and TL2 as the detection areas provided in the membrane M and the control line CL as the operation check area are directly exposed to the outside without being covered by the cover tape 5. Can do. Thus, when the user confirms the presence or absence of the line on the test lines TL1, TL2 and the control line CL, the test strip 2 is not obstructed by the cover tape 5, and the test lines TL1, TL2 and the control line CL The presence or absence of line expression in can be reliably confirmed.

  In this embodiment, the cover tape 5 also covers the surface of the sample pad side laminated portion 8a on which the sample pad SP and the conjugate pad CP are laminated, and the membrane M and the membrane on which the conjugate pad CP is laminated. The side laminated portion 8b is also formed so as to cover it. Since the cover tape 5 is a soft member with an adhesive member formed on the back surface, it conforms to the surface shape of the sample pad SP, conjugate pad CP, sample pad side laminated portion 8a, and membrane side laminated portion 8b. The sample pad SP, the conjugate pad CP, the sample pad side laminated portion 8a, and the membrane side laminated portion 8b can be attached in close contact with each surface without forming a gap. Further, since the cover tape 5 is formed of a transparent member, there is air or dust between the sample pad SP, the conjugate pad CP, the sample pad side laminated portion 8a, and the membrane side laminated portion 8b. Whether or not can be visually confirmed from the outside.

  Incidentally, in the membrane M, two test lines TL1 and TL2 and a control line CL are arranged as a detection region and an operation confirmation region so as to extend in the width direction y orthogonal to the deployment direction x. The specimen extract that develops in the development direction x can pass through the test lines TL1 and TL2 and the control line CL sequentially. Thereby, in the membrane M, as in the conventional case, when an antigen is present in the sample extract, a complex produced by a specific reaction between the labeled antibody of the conjugate pad CP and the antigen is produced. For example, when passing through the test line TL1, it reacts specifically with the antigen-capturing antibody, and the line can be expressed in the test line TL1 by the fluorescent substance contained in the labeled antibody of the complex. Further, in the membrane M, when an unreacted labeled antibody passes through the control line CL, the labeled antibody reacts specifically with the labeled capture antibody so that the line can be expressed in the control line CL.

  Here, the test strip 2 is covered with the cover tape 5 so as to cover each surface from the sample pad SP to the end of the conjugate pad CP via the sample pad-side laminated portion 8a, so that the sample pad SP The area covered with the cover tape 5 while blocking the specimen extract that is about to flow along the conjugate pad CP from the sample pad SP when the specimen extract is dropped on the cover tape 5 Thus, the specimen extract can be guided along the cover tape 5 extending in the development direction x to the optimum path for development from the sample pad SP to the conjugate pad CP via the sample pad side laminated portion 8a.

  Thus, the test strip 2 increases the probability that the labeled antibody and the antigen in the specimen extract will specifically react, and the complex obtained by specifically reacting the labeled antibody and the antigen is tested by the Membrane M test. It is urged to be deployed to the lines TL1, TL2 and the control line CL, and the complex can be quickly led to the test lines TL1, TL2 and the control line CL.

(1-2) Action and Effect In the above configuration, in the test strip 2, the antigen capture antibody is immobilized while covering the entire surface of the conjugate pad CP from the non-exposed region of the sample pad SP via the sample pad side laminated portion 8a. The cover tape 5 for exposing the membrane M provided with the test lines TL1, TL2 and the control line CL, which are the detection areas, to the outside is provided. As a result, in the test strip 2, the specimen extract is guided to the optimum path to be developed from the sample pad SP into the conjugate pad CP through the sample pad side laminated portion 8a along the cover tape 5 extending in the development direction x. The probability that the labeled antibody reacts specifically with the antigen in the specimen extract is increased, making it easier to obtain a complex.

  As a result, test strip 2 can capture more complex antigens with antigen-capturing antibodies, which can promote the expression of test lines TL1 and TL2 than before, and the complex can be obtained quickly. As a result, the time until the antigen-capturing antibody captures the antigen of the complex can be shortened. Thus, it can be easily determined in a shorter time than before whether or not the specimen is present in the specimen extract. .

  In addition, the test strip 2 is provided with a cover tape 5 even if a sample extract exceeding a predetermined amount (for example, about 2 drops (about 55 μL)) is accidentally dropped on the sample pad SP. In this way, the specimen extract can be guided to the optimum path for developing from within the sample pad SP covered with the cover tape 5 into the conjugate pad CP via the sample pad side laminated portion 8a. Therefore, in the test strip 2, the probability that the labeled antibody and the antigen in the specimen extract will specifically react will be higher than before, and it will be easier to obtain the complex. Since the body antigen can be captured, line expression of the test lines TL1 and TL2 can be promoted more than before, and the complex can be obtained quickly, until the antigen capture antibody captures the complex antigen. Can be shortened.

  In this test strip 2, the cover tape 5 is not attached to the membrane M provided with the test lines TL1 and TL2, which are detection areas where the antigen-capturing antibody is immobilized, and the control line CL as an operation confirmation area. Because it is exposed to the outside, the user can directly see the test lines TL1, TL2 and the control line CL without being obstructed by the cover tape 5, and the line expression in the test lines TL1, TL2 and the control line CL. The presence or absence of can be easily determined.

  Furthermore, in this test strip 2, the cover tape 5 was stuck to the end of the conjugate pad CP that became the boundary with the membrane M without sticking the cover tape 5 to the membrane M. As a result, in the test strip 2, the sample extract is spread from the conjugate pad to the membrane without forming a gap between the step portion between the membrane M and the membrane-side laminated portion 8b and the cover tape 5. At this time, the occurrence of convection due to the gap can be suppressed, and the occurrence of uneven flow of the specimen extract can be prevented.

  In the case of this embodiment, in the test strip 2, the specimen extract dropped onto the sample pad SP is blocked by the thickness of the cover tape 5 at one end of the cover tape 5, and the specimen extract sample Absorption to the pad SP can also be promoted. Furthermore, since the convex stepped portion that is the joint between the sample pad side laminated portion 8a and the conjugate pad CP is also covered with the cover tape 5, the analyte extract absorbed in the sample pad SP is used for the conjugate pad. CP can be reliably developed, and accordingly, the probability that the labeled antibody and the antigen in the specimen extract react specifically can be increased.

  According to the above configuration, even if the user drops a sample extract of a specified amount or more onto the sample pad SP, the sample extract is guided into the conjugate pad CP by the cover tape 5, and the labeled antibody and the sample extract are introduced. It is easy to obtain a complex by specifically reacting with the antigen in the membrane, and as a result, a larger amount of the antigen of the complex can be captured by the membrane M antigen capture antibody. The time until the antigen is captured can be shortened, and thus whether or not a sample is present in the sample extract can be easily determined in a shorter time than before.

(2) Second Embodiment In the above-described embodiment, as the cover tape, a cover tape that covers from the unexposed region of the sample pad SP to the membrane side laminated portion 8b that overlaps the membrane M of the conjugate pad CP. Although the test strip 2 provided with 5 has been described, the present invention is not limited to this, and the same reference numerals are assigned to the corresponding parts in FIG. 3A and FIG. As shown in FIG. 3B, the test strip 12 may be provided with a cover tape 15 that covers from the non-exposed region of the sample pad SP to beyond the membrane-side laminated portion 8b and further to a partial region of the membrane M.

  In this case, the cover tape 15 shown in FIG. 3A and FIG. 3B differs from the cover tape 5 shown in FIG. 2A and FIG. The following description will focus on the other end of the cover tape 15. In this case, the cover tape 15 exceeds the membrane side laminated portion 8b in which the membrane M and the conjugate pad CP are laminated, and the other end is arranged in a region that does not reach the test line TL1 of the membrane M, and the sample pad SP The membrane M is pasted so as to cover from the non-exposed region to a partial region of the membrane M through the conjugate pad CP. Thus, although the cover tape 15 is applied to a part of the membrane, the test line TL1, TL2 and the control line CL are externally covered without covering the test line TL1, TL2 and the control line CL of the membrane M. It is made to be exposed. Even with such a configuration, it is possible to obtain the same effect as that of the test strip 2 according to the first embodiment described above.

  That is, in this test strip 12 as well, the specimen extract is guided to the optimum path for development from the sample pad SP through the sample pad side laminated portion 8a into the conjugate pad CP along the cover tape 15 extending in the development direction x. The probability that the labeled antibody reacts specifically with the antigen in the specimen extract is increased, making it easier to obtain a complex.

  As a result, the test strip 12 can capture more complex antigens with the antigen-capturing antibody, so that the line expression of the test lines TL1 and TL2 can be promoted more than before, and the complex can be obtained quickly. As a result, the time until the antigen-capturing antibody captures the antigen of the complex can be shortened. Thus, it can be easily determined in a shorter time than before whether or not the specimen is present in the specimen extract. .

  Also, the test strip 12 is provided with a cover tape 15 even if a sample extract exceeding a predetermined amount (for example, about 2 drops (about 55 μL)) is accidentally dropped on the sample pad SP. As a result, the specimen extract can be guided to the optimal path that develops from the sample pad SP covered by the cover tape 15 into the conjugate pad CP via the sample pad side laminated portion 8a. Therefore, in the test strip 12, the probability that the labeled antibody and the antigen in the specimen extract will specifically react will be higher than before, and it is easier to obtain the complex. Since the body antigen can be captured, line expression of the test lines TL1 and TL2 can be promoted more than before, and the complex can be obtained quickly, until the antigen capture antibody captures the complex antigen. Can be shortened.

  Even in this case, in the test strip 12, the test lines TL1 and TL2 as the detection areas provided in the membrane M and the control lines CL as the operation check areas are not directly covered with the cover tape 15 but exposed to the outside. Therefore, when confirming the presence or absence of line expression on the test lines TL1, TL2 and the control line CL, the line expression on the test lines TL1, TL2 and the control line CL is not obstructed by the cover tape 15. The presence or absence of can be confirmed directly.

(3) Other Embodiments In the above-described embodiment, the test strip 2 in which a part of the sample pad SP is covered with the cover tape 5 from the end of the conjugate pad CP that becomes the boundary with the membrane M Although the test strip 12 in which a part of the sample pad SP is covered with the cover tape 15 from the part of the sample pad SP to the part of the membrane M has been described, the present invention is not limited to this. As shown in FIG. 4A, the corresponding parts of FIG. 4A are covered with the cover tape 25 from the membrane side laminated portion 8b to a partial region of the conjugate pad CP without covering the sample pad SP. As shown in FIG. 4B in which the same reference numerals are assigned to the corresponding parts to FIG. A test strip 32 Tsu may be applied.

  In this case, the test strip 22 shown in FIG. 4A covers the entire area of the membrane M and the sample pad SP while covering the area from the boundary between the membrane M and the conjugate pad CP to a predetermined region of the conjugate pad CP. 25 is provided, and even with such a cover tape 25, the same effects as those of the above-described embodiment can be obtained.

  In practice, this cover tape 25 is applied over the membrane side laminated portion 8b in which the membrane M and the conjugate pad CP are laminated, and the region of about two-thirds of the conjugate pad CP. The sample pad side laminated portion 8a in which the gate pad CP and the sample pad SP are laminated can be exposed to the outside without covering.

  On the other hand, the test strip 32 shown in FIG. 4B covers the membrane side laminated portion 8b and the membrane M while covering a partial region of the sample pad SP to a partial region of the conjugate pad CP beyond the sample pad side laminated portion 8a. A cover tape 35 that exposes the entire surface of the cover tape 35 to the outside is provided. Even with such a cover tape 35, the same effects as those of the above-described embodiment can be obtained.

  In practice, the cover tape 35 is formed from the unexposed region of the sample pad SP covered by the lid 102 (FIG. 1) of the sample pad SP, beyond the sample pad side laminated portion 8a, to about the conjugate pad CP. It is affixed over a two-thirds region so that it can be exposed to the outside without covering the membrane-side laminated portion 8b.

  As other embodiments, the sample pad side laminated portion 8a and the test line of the membrane M are covered from the partial region of the conjugate pad CP to the partial region of the membrane M beyond the membrane side laminated portion 8b. You may apply the cover tape exposed outside, without covering TL1 and the control line CL.

  Incidentally, in the test strips 2 and 12 shown in FIG. 2A and FIG. 3A, two test lines TL1 and TL2 are provided as detection areas. However, the present invention is not limited to this, and one or more test lines TL1 are provided. You may do it. Further, in the test strips 22 and 32 shown in FIG. 4A and FIG. 4B, one test line TL1 is provided as a detection region, but the present invention is not limited to this, and two or more test lines may be provided.

  In the above-described embodiment, an antigen is applied as a specimen, a labeled antibody is applied as a labeling substance having a fluorescent substance, an antigen capturing antibody and a labeled capturing antibody are applied as a capturing substance, and an antigen-antibody reaction is performed. Although the immunochromatographic test strip used has been described, the present invention is not limited to this, and various other specimens, labeling substances, and capture substances can be applied to use, for example, complementary nucleic acid reactions or ligand-receptor reactions. It may be a test strip for chromatography. Furthermore, for example, a test strip having no absorption pad may be used as long as the specimen extract can be developed in order from the sample pad to the conjugate pad and the membrane.

  Furthermore, in the above-described embodiment, the case where the cover tapes 5 and 15 made of the soft adhesive tape attached to the test strips 2 and 12 are applied as the cover member is described, but the present invention is not limited to this. For example, a cover member made of a hard plate member attached to the test strips 2 and 12 may be applied.

  Further, in the above-described embodiment, the case where the cover tapes 5 and 15 attached to the test strips 2 and 12 are applied as the cover members has been described. However, the present invention is not limited to this, for example, on the case body side. A cover member may be provided, and at least the conjugate pad may be covered with the cover member of the case body between the conjugate pad of the test strip and the test line TL1 of the membrane.

  In this case, in the test kit, when the lid is fixed to the base on which the test strip is installed, the area of the conjugate pad or the like to be covered (cover tape 5 in FIG. 2 or FIG. A cover member may be provided in advance in a region facing the region covered with the cover tape 15. Even in such a test kit, the specimen extract can be guided to the optimal path for deployment from the sample pad SP into the conjugate pad CP along the cover member extending in the deployment direction x, as in the above-described embodiment. The effect of can be obtained.

  In the above-described embodiment, colored latex particles, metal sol (gold colloid, etc.), fluorescent latex particles, magnetic beads, enzymes (alkaline phosphatase, etc.) may be applied as the labeling substance. When, for example, colored latex particles or gold colloid is used as the labeling substance, the user can visually confirm the presence or absence of line expression on the test lines TL1 and TL2, while fluorescent labeling particles are used as the labeling substance. When used, the presence or absence of line expression on the test lines TL1 and TL2 can be confirmed with a fluorescence reader or the like.

  Furthermore, as another embodiment, for example, a test strip used for detection of influenza virus may be used. Such a test strip for influenza virus has the same basic structure as the test strips 2, 12, 22, 32, 42, 52 described above, but the labeling substance held in the conjugate pad CP and the membrane M The capture substance immobilized on the detection region may be appropriately changed according to the influenza virus to be detected.

  In fact, the test strip for influenza virus has a conjugate pad CP in which a labeling substance that specifically reacts with the influenza virus to form a complex (labeling substance for influenza virus) is held releasably in a wet state. And a membrane M having a detection region on which a capture substance (capture substance for influenza virus) that specifically reacts with a complex developed by capillary action is immobilized.

  For example, a test strip that can detect both influenza A and influenza B viruses, an influenza virus A colored latex-labeled antibody that specifically reacts with influenza A virus to form a complex, and influenza B An influenza virus type B colored latex-labeled antibody that specifically reacts with the virus to form a complex is releasably retained in the conjugate pad CP in a wet state. In addition, the conjugate pad CP holds a labeling substance having a different color depending on the number of influenza viruses to be detected. In this case, for example, the influenza virus type A colored latex labeled antibody is blue, and the influenza virus type B The colored latex labeled antibody is colored red.

  In the test strip for influenza virus, test lines TL1 and TL2 as detection areas are provided in the membrane M. For example, an influenza A virus antibody (for example, a mouse-derived anti-influenza virus A type lgG is provided on one test line TL1. Monoclonal antibody), and the other test line TL2 contains an influenza B virus antibody (for example, an anti-influenza virus B IgG monoclonal antibody). The membrane M may be provided with a control line CL on which an antigen capture antibody (for example, goat-derived anti-mouse IgG polyclonal antibody) is immobilized at a position downstream of the test lines TL1 and TL2.

  In such a test strip for influenza virus, a cover tape covering at least the conjugate pad CP is provided between the conjugate pad CP and the detection region of the membrane M (test lines TL1, TL2). As described above, the antigen capture antibody can capture a larger amount of the antigen of the complex, so that the line expression of the test lines TL1 and TL2 can be promoted more than before, and the complex can be obtained quickly. As a result, the time required for the antigen-capturing antibody to capture the antigen of the complex can be shortened. Thus, it is easier to determine whether or not influenza virus is present in the sample extract in a shorter time than before. obtain.

(4) Verification Test (4-1) Test Strip Using Blue-colored Latex Particles Next, a conventional test strip not provided with a cover tape is different from the pasting range of the cover tape, FIGS. 4A, 4B, and 4C. And 4 types of test strips shown in FIG. 4D and test strips that cover not only the conjugate pad CP but also the entire surface of the membrane M are prepared. The detection time until the line appeared in the line TL1 and the control line CL was examined. Note that the test strip used in this verification test differs only in the cover tape configuration, and the sizes and materials of the support material 111, sample pad, conjugate pad, membrane (here, nitrocellulose membrane), and absorbent pad are used. Were the same.

  First, a verification test was performed using an inspection kit (also called an adeno plate) in which an adeno test strip was installed in the case body. In addition, the test strip for adeno was produced as follows. First, after preparing a nitrocellulose membrane (HF90 membrane, manufactured by Merck Millipore), a mouse-derived anti-adenovirus IgG monoclonal antibody was dissolved in PBS, at a concentration of 1.0 mg / ml at a rate of 1.0 μl / cm, Test line TL1 was produced by applying in a line on the upstream side in the developing direction when viewed from the end of the membrane with a positive pressure spray device Biojetxyz3060 (Nippon Biodot). Separately, goat-derived anti-mouse IgG polyclonal antibody is dissolved in PBS, and at a concentration of 1.0 mg / ml at a rate of 1.0 μl / ml, by a positive pressure spray device Biojetxyz3060 (Nippon Biodot), The control line CL was produced by applying in a line on the downstream side of the test line TL1 in the development direction when viewed from the end of the membrane.

  The antibody-coated membrane is placed in a dryer at 40 ° C and allowed to dry overnight. After that, the membrane is immersed in a solution containing 1% BSA (bovine serum albumin), 0.1% skim milk, and a surfactant for 30 minutes. After blocking the region, the membrane M was prepared by drying overnight in an oven.

  Subsequently, a blue colored latex particle-labeled antibody suspension was prepared as follows. First, the carboxyl group of the blue colored latex particles was activated with the same amount of water-soluble carbodiimide (1-Ethyl-3- (3-dimethylaminopropyl) carbodiimide, hydrochloride, Wako Pure Chemical Industries) in a test tube. By adding 62.5 mg of anti-adenovirus monoclonal antibody (1.Omg / ml) per 1 g of activated blue colored latex particles, mixing well, and stirring with a stirrer at 4 ° C overnight Reacted. After completion of the reaction, the mixture was centrifuged at 20,000 G for 10 minutes, and then the blue colored latex particle-labeled antibody from which the supernatant had been removed was probed with a buffer containing 0.1% BSA, and after completion, 20,000 G For 10 minutes, and the supernatant was removed. Thereafter, a predetermined amount of 5% sucrose water containing 1% BSA was added and resuspended with a pipette to obtain a blue colored latex particle-labeled antibody suspension.

  Prepare a blue colored latex particle labeled antibody suspension by adding a predetermined amount of 4% sucrose water containing 1% BSA to the resulting blue colored latex particle labeled antibody suspension and resuspending it with a pipette. Apply this blue colored latex particle-labeled antibody suspension at a rate of about 45 μl per cm to an 8 mm long conjugate pad (glass fiber, manufactured by Merck Millipore), and spread it evenly in an oven. And dried to make a conjugate pad CP.

  Next, the prepared membrane M was bonded to a backing sheet (Adhesives Research, Inc) serving as the support material 111, and then the prepared conjugate pad CP was bonded to the membrane M so as to overlap with the membrane M by several millimeters. Further, on the upstream side in the development direction, a glass fiber (manufactured by Merck Millipore) was bonded as a sample pad SP so as to overlap the conjugate pad CP by several millimeters. On the downstream side of the membrane M, an absorption pad AbP (sample absorption pad, manufactured by Merck Millipore) for receiving excess reagent is arranged to overlap the membrane M several millimeters, and the test strip before applying the cover tape Was made.

  The cover tape to be applied to each test strip is a polyester film (model number PET25 (A) PL Thin 8LK, manufactured by Nippon Biodot Co., Ltd.), sheared to a predetermined length, and FIGS. 4A, 4B, 4C and 4D. It stuck on each position shown in.

  In this case, a sample pad SP having a longitudinal dimension of 13 mm extending in the developing direction x and a conjugate pad CP having a longitudinal dimension of 8 mm extending in the developing direction x are used, as shown in FIG. The length x2 of the sample pad side laminated portion 8a in which the SP and the conjugate pad CP are laminated is set to 1 mm, and the length x4 of the membrane side laminated portion 8b in which the membrane M and the conjugate pad CP are laminated is set to 2 mm. .

  The test strip 22 shown in FIG. 4A (hereinafter also simply referred to as a 5 mm first cover tape) uses a cover tape 25 extending in the deployment direction x and having a longitudinal dimension of 5 mm, and the membrane M and the conjugate pad CP. Cover tape 25 was applied from the boundary to the surface of conjugate pad CP. Specifically, as shown in FIG. 5, the distance x3 from one end of the conjugate pad CP to one end of the sample pad CP was set to 2 mm, and the conjugate pad CP in the region of the distance x3 was exposed to the outside.

  The test strip 32 shown in FIG. 4B (hereinafter also simply referred to as a 5 mm second cover tape) uses a cover tape 35 having the same longitudinal dimension extending in the developing direction x, but a partial region of the sample pad SP. To the surface of the conjugate pad CP. Specifically, as shown in FIG. 5, a distance x1 from one end of the conjugate pad CP in the sample pad side laminated portion 8a to one end of the cover tape 35 on the sample pad SP is set to 1 mm. The length of the conjugate pad CP to be covered was 3 mm.

  The test strip 42 shown in FIG. 4C (hereinafter, also simply referred to as a 9 mm cover tape) uses the cover tape 5 having a longitudinal dimension of 9 mm extending in the development direction x, and a conjugate pad from a partial region of the sample pad SP. The entire surface of the CP was covered with the cover tape 5. Specifically, as shown in FIG. 5, the distance x1 from one end of the conjugate pad CP in the sample pad side laminated portion 8a to one end of the cover tape 35 on the sample pad SP is 1 mm, and the conjugate pad CP Was covered with cover tape 5.

  The test strip 52 shown in FIG. 4D (hereinafter also simply referred to as “12 mm cover tape”) uses the cover tape 15 having a longitudinal dimension of 12 mm extending in the deployment direction x, and is a conjugate pad from a partial region of the sample pad SP. A part of the membrane M was covered with the cover tape 15 through the CP. Specifically, as shown in FIG. 5, the distance x1 from one end of the conjugate pad CP in the sample pad side laminated portion 8a to one end of the cover tape 15 on the sample pad SP is 1 mm, and the conjugate pad The distance x5 from the other end to the other end of the cover tape was 3 mm.

  As a comparative example, as shown in FIG. 5, the distance x1 from one end of the conjugate pad CP in the sample pad side laminated portion 8a to one end of the cover tape 117 on the sample pad SP is 1 mm, and the conjugate pad A test strip (hereinafter also simply referred to as a full-surface cover tape) with a cover tape 117 that covers the entire surface of the CP and the membrane M was prepared.

  Next, the effectiveness of the cover tapes 5, 15, 25, 35, and 117 was examined using an adenovirus immunochromatography test apparatus. As a specimen, 90 μL of antigen prepared at a predetermined concentration (Adenovirus CF Reagent “SEIKEN”, DENKA SEIKEN) was used. Ten test strips were prepared, and each test strip was installed in the case body to prepare multiple test kits. Next, place each test kit horizontally on a flat surface on a desk so that the sample drop opening of the test kit is located on the upper side, and then pass the positive sample with a specified amount of pipette to the sample drop opening of each test kit. The reaction time of each test strip (line detection time) and the reaction state were observed.

  Specifically, the time when the control line and the test line appeared in the determination opening of each test kit was measured as the detection time, and the flow of the solution in each test strip was observed. In addition, 5 minutes after dropping the positive sample, the test kit is destroyed and the sample in the case body is removed to stop the reaction on the test strip, and the line intensity of the line that appears on the test line is determined by the immunochromatography reader. (Measured by Hamamatsu Photonics).

  As a result, in the test strip without the cover tape as a comparative example, the result as shown in FIG. 6 was obtained. In this test strip without cover tape, the average value of 10 lines of strength is 35.3 mAbs, and the average value of detection time of 10 lines (expressed as T line in the table) on the test line is 38.7 sec. Thus, the average value of the detection time of 10 lines in the control line (denoted as C line in the table) was 36.4 sec. Furthermore, when the flow of the solution on the test strip without the cover tape and the cleanliness of the line were visually confirmed, no particular change of state was noticed.

  Further, in the test strip of the full cover tape as another comparative example, the result as shown in FIG. 7 was obtained. From FIG. 7, in the test strip of the full surface cover tape, the line strength was lowered and the detection time of the C line was longer than that without the cover tape. Furthermore, on the test strip of the full surface cover tape, when the flow of the solution and the cleanliness of the line were visually confirmed, a streak-like flow such as smoke was generated, and unevenness was confirmed in the flow. Further, the solution from the membrane was not easily removed, and the color of the blue colored latex particles remained on the membrane even after 5 minutes had passed from the dropping of the positive specimen, making it difficult to see the line on the membrane.

  On the other hand, with the test strip 22 of the 5 mm first cover tape of the present invention, the results shown in FIG. 8 were obtained. From Fig. 8, the test strip 22 of the 5mm first cover tape has an average value of 10 line strengths compared with the test strip without the cover tape as a comparative example and the test strip with the full cover tape as a comparative example. Was 48.8 mAbs, and it was confirmed that the line strength was higher than that of the comparative example and the line was easier to see. Moreover, the detection time of the T line was also shorter than that of the comparative example, and it was confirmed that the presence or absence of the line could be determined in a short time. Furthermore, with the test strip 22 of this 5mm first cover tape, when the flow of the solution and the cleanliness of the line were visually confirmed, the flow of the solution flowed evenly with no unevenness, and the line was clearly expressed. There was no particular change in condition.

  Further, in the test strip 32 of the 5 mm second cover tape of the present invention, the result as shown in FIG. 9 was obtained. From FIG. 9, the test strip 32 of the 5 mm second cover tape has an average value of 10 line strengths compared with the test strip without the cover tape as a comparative example and the test strip with the full cover tape as a comparative example. Was 47.4 mAbs, and it was confirmed that the line strength was higher than that of the comparative example and the line was easier to see. Moreover, the detection time of the T line and C line was also shorter than that of the comparative example, and it was confirmed that the presence or absence of the line could be determined in a short time. Furthermore, with the test strip 32 of this 5mm second cover tape, when the flow of the solution and the cleanliness of the line were visually confirmed, the flow of the solution flowed evenly with no unevenness, and the line was clearly expressed. There was no particular change in condition.

  Further, in the test strip 42 of the 9 mm cover tape of the present invention, the result as shown in FIG. 10 was obtained. From FIG. 10, the test strip 42 of 9 mm cover tape has an average value of 10 line strengths of 47.6 in comparison with the test strip without the cover tape as a comparative example and the test strip with the full cover tape as a comparative example. It was confirmed that the line strength was higher than that of the comparative example and the line was easier to see than the comparative example. Moreover, the detection time of the T line and C line was also shorter than that of the comparative example, and it was confirmed that the presence or absence of the line could be determined in a short time. Furthermore, in this test strip 42 of the 9mm cover tape, when the flow of the solution and the cleanliness of the line were visually confirmed, the flow of the solution flowed evenly without unevenness, and the line was clearly expressed, There was no particular change in condition.

  Further, with the test strip 52 of the 12 mm cover tape of the present invention, results as shown in FIG. 11 were obtained. From FIG. 11, the 12 mm cover tape test strip 52 has an average value of 10 line strengths of 48.6 compared with the test strip without the cover tape as a comparative example and the test strip with the full cover tape as a comparative example. It was confirmed that the line strength was higher than that of the comparative example and the line was easier to see than the comparative example. Moreover, the detection time of the T line and C line was also shorter than that of the comparative example, and it was confirmed that the presence or absence of the line could be determined in a short time. However, in the test strip 52 of this 12 mm cover tape, when the flow of the solution and the cleanliness of the line were visually confirmed, there was an unevenness in the flow of the solution, and there were also unevenness in the expression state of the line. there were. Furthermore, some membrane solutions were poorly removed, and even after 5 minutes had passed since the positive specimen was dropped, the color of the blue colored latex particles remained on the membrane, making it difficult to see the line on the membrane.

  From this, in addition to increasing the line strength and shortening the detection time, in addition to considering how the solution flows on the test strip and maintaining the cleanliness of the line, rather than using 12 mm cover tape, It was confirmed that it is desirable to use 5mm first cover tape, 5mm second cover tape, and 9mm cover tape.

  Next, the amount of specimen dropped onto the sample pad was changed to 40 μL, 80 μL, 120 μL, and 160 μL, and the test strip reaction time (detection time) and reaction state were observed in the same manner as described above. First, in this verification test, a test strip without a cover tape and a test strip with a full cover tape were used as comparative examples.

  In this verification test, the test strip used in the test kit of the present invention includes a test strip using a 5 mm first cover tape (FIG. 4A) and a test strip using a 5 mm second cover tape (FIG. 4B). A test strip (FIG. 4C) using a 9 mm cover tape was used.

  Each test strip was installed in the case body to prepare a plurality of test kits. Next, change the sample volume of the positive sample (Adenovirus CF reagent “SEIKEN”, DENKA SEIKEN) dropped on the test strip to 40 μL, 80 μL, 120 μL, and 160 μL, and 10 test strips for each of these different sample volumes. The line detection time and reaction state were observed.

  Specifically, a positive sample of the above-described sample amount is dropped with a pipette into the sample dropping opening of each test kit, and the time when the control line and the test line appear in the determination opening of each test kit is determined. The detection time was measured, and the flow of the solution in each test strip was observed. In addition, 5 minutes after dropping the positive sample, the test kit is destroyed and the sample in the case body is removed to stop the reaction on the test strip, and the line intensity of the line that appears on the test line is determined by the immunochromatography reader. (Measured by Hamamatsu Photonics).

  FIG. 12 is a graph summarizing the average values of the line intensities measured for each test strip when the sample amount is 40 μL, 80 μL, 120 μL, and 160 μL. In the following, including FIG. 12, the test strip without the cover tape, which is a comparative example, is also referred to as “no tape”, and the test strip of the full cover tape, which is also a comparative example, is also referred to as “full tape”. The test strip of the example using the 5mm first tape is also called the "5mm second tape" and the test strip of the example using the 5mm second cover tape is also called the "5mm second tape" of the example using the 9mm cover tape. The test strip is also called “9mm tape”.

  From FIG. 12, in the test strip as a comparative example without the cover tape, the line strength of the test line that was 41 mAbs when the sample volume was 40 μL became less reactive as the liquid volume increased, and the sample volume was 120 μL. In the case of, the line strength decreased to about 1/3 compared with 40 μL. From this, it was confirmed that in the conventional test strip without the cover tape, when the sample amount exceeds the specified amount, the line strength of the test line is lowered, and it is difficult for the user to make an accurate judgment.

  On the other hand, the 5 mm first tape, the 5 mm second tape, and the 9 mm tape showed an effect of suppressing a decrease in line strength accompanying an increase in the amount of specimen. It was also found that the effect of suppressing the decrease in line strength was the largest for the 9 mm tape and the second for the 5 mm first tape. On the other hand, with the full-surface tape as a comparative example, the sample could not be completely removed from the membrane M even after all of the excess sample flowed, and the effect of suppressing a decrease in line strength due to an increase in the amount of sample added was not seen. .

  In addition, the results shown in FIG. 13 were obtained for the test line detection time on the test strip when the sample amount was changed. FIG. 13 is a graph summarizing the average values of the detection times at which lines were developed on the test line (T line) for each test strip when the sample volume was 40 μL, 80 μL, 120 μL, and 160 μL. From FIG. 13, without the tape as a comparative example, a delay in the line detection time on the test line was recognized as the sample amount increased.

  On the other hand, with 5mm 1st tape, 5mm 2nd tape, and 9mm tape, when adding 120μL or more, which is more than the normal specified amount, the detection time of the line on the test line is shorter than without the tape of the comparative example. It was confirmed that there was an effect of delaying the detection time of the line. On the other hand, in the full cover as a comparative example, the delay preventing effect was not confirmed as compared with the case without the tape. For this reason, when a sample extract of a specified amount or more is added, shortening the detection time on the test line cannot be achieved simply by applying a cover tape to the entire conjugate pad or membrane. It was confirmed that it was necessary to apply the cover tape to a specific range including the conjugate pad portion like the 5 mm first tape, 5 mm second tape and 9 mm tape. It was also found that the line detection time could be shortened most especially with 9mm tape.

  Based on the above, when a specimen extract of a specified amount or more is dripped onto a test kit due to human error at a clinical site, if the specimen is very weak with a small amount of antigen, a conventional test strip without tape or The test strip with the cover tape attached to the entire surface may be judged negative even though it is originally positive. In contrast, in the test strip of the example in which the cover tape is applied to a specific range including the conjugate pad portion such as the 5 mm first tape, the 5 mm second tape, and the 9 mm tape, the sample extraction exceeding the specified amount is temporarily assumed. Even if the liquid is dropped, the line strength at the test line is more clearly expressed than before, so that the false negative determination can be avoided as compared with the conventional case.

  Next, for the test strip (FIG. 4D) using the 12 mm cover tape, the amount of the sample dropped on the sample pad was changed to 40 μL, 80 μL, 120 μL, and 160 μL. The detection time and reaction state were observed. Here, in order to verify the detection time of the line in the test strip using the 12 mm cover tape and the reaction state, the test strip without the cover tape as a comparative example and the test strip of the example using the 9 mm cover tape ( The verification test was performed again for FIG. 4C).

  Also in this verification test, 40 μL, 80 μL, 120 μL, and 160 μL of each positive sample (Adenovirus CF reagent `` SEIKEN '', DENKA SEIKEN) was pipetted with the sample drop opening of each test kit as above. The detection time when the control line and the test line appeared in the determination opening of each test kit was measured, and the flow of the solution in each test strip was observed. In addition, 5 minutes after dropping the positive sample, the test kit is destroyed and the sample in the case body is removed to stop the reaction on the test strip, and the line intensity of the line that appears on the test line is determined by the immunochromatography reader. (Measured by Hamamatsu Photonics).

  FIG. 14 is a graph summarizing the average values of the line intensities of the test lines measured for each test strip when the sample amount is 40 μL, 80 μL, 120 μL, and 160 μL. In the following, including FIG. 14, the test strip of the example using the 12 mm cover tape is also referred to as “12 mm tape”. The results for “no tape” and “9 mm tape” shown in FIG. 14 are different from the results shown in FIG. 12, but this is due to differences in ambient conditions such as humidity and temperature when the verification test was performed. Is due to.

  From FIG. 14, in the test strip as a comparative example without the cover tape, the line strength in the test line having 35 mAbs when the sample volume is 40 μL becomes less reactive as the liquid volume increases, and the sample volume becomes smaller. At 120 μL, the line strength was reduced to about 1/3 compared with 40 μL, which was the same as the result of FIG. On the other hand, the 12 mm tape was also effective in suppressing the decrease in line strength accompanying the increase in the amount of specimen, similar to the 9 mm tape.

  Further, the test line detection time on the test strip when the sample amount was changed was obtained as shown in FIG. FIG. 15 is a graph summarizing the average values of detection times when a line (T line) appears on the test line for each test strip when the sample amount is 40 μL, 80 μL, 120 μL, and 160 μL. From FIG. 15, without the tape as a comparative example, the delay of the line detection time on the test line was recognized as the sample amount increased, but the 12 mm tape, like the 9 mm tape, 120 μL which exceeds the normal specified amount. When added as described above, the line detection time on the test line was shorter than that of the comparative example without the tape, and it was confirmed that there was an effect of preventing the delay of the line detection time. From the above, even with test strips using 12mm tape, if more than the specified amount of specimen extract is dripped, the line strength at the test line is more clearly expressed than before, so it is more false negative than before. Can be avoided.

(4-2) Test strip using colloidal gold particles Next, the effect of shortening the detection time of the line on the test line by applying the cover tape and the effect of preventing the delay of the detection time of the line are group A β-streptococci (strept A verification test was performed to confirm whether or not A) was obtained even when the sample was used as a specimen or when a metal sol was used as a labeling substance. Note that the test strip used in this verification test differs from the verification test described above only in that gold colloid particles are used as a labeling substance. Similar to the verification test described above, The test strip 22 of the 5 mm first cover tape, the test strip 32 of the 5 mm second cover tape, the test strip 42 of the 9 mm cover tape, and the test strip 52 of the 12 mm cover tape having the configuration shown in FIG.

In this case, a verification test was performed using an inspection kit (also referred to as a strep A plate) in which a test strip for strep A was installed in the case body. The test strip for Strep A was produced as follows. Here, group A β-streptococcus prepared at a concentration of 8 × 10 ⁵ cells / mL was used as a positive sample. First, a test strip before applying a cover tape for detecting the antigen of this group A β-streptococcus was prepared according to the following procedure. In this case, after preparing a nitrocellulose membrane (HF90 membrane, manufactured by Merck Millipore), a rabbit-derived anti-Group A β-streptococcal glycan antigen F (ab ′) 2 antibody was added at 1.0 μl / cm at a concentration of 1.0 mg / ml. The test line TL1 was produced by applying in a line shape on the upstream side in the development direction when viewed from the end of the membrane with a positive pressure spray device Biojetxyz3060 (Nihon Biodot Co., Ltd.). Separately, goat-derived anti-mouse IgG polyclonal antibody was dissolved in PBS, and the membrane was added at a concentration of 1.0 mg / ml at a rate of 1.0 μl / ml using a positive pressure spray device Biojetxyz3060 (Japan Biodot). The control line CL was produced by coating in a line on the downstream side of the test line TL1 in the development direction as viewed from the end of the test line.

  The membrane coated with the antibody was placed in a dryer at 40 ° C and allowed to dry overnight, and then immersed in a PBS solution containing 1% BSA (bovine serum albumin) and a surfactant for 30 minutes to remove the protein unadsorbed area. After blocking, the membrane M was produced by drying overnight in an oven.

  Next, add rabbit-derived anti-group A β-streptococcus glycan antigen IgG antibody (1.0 mg / mg) to 30 volumes of colloidal gold solution in a test tube at a ratio of 1 volume, mix well, and then mix at 4 ° C. The reaction was carried out by stirring with a stirrer in the evening. After completion of the reaction, the mixture was centrifuged at 20,000 G for 10 minutes, and then a predetermined amount of 4% sucrose water containing 1% BSA was added to the gold colloid-labeled antibody from which the supernatant had been removed. A colloid-labeled antibody suspension was obtained.

  Then, apply this gold colloid-labeled antibody suspension at a rate of about 45 μL per 1 cm to an 8 mm long conjugate pad (glass fiber, manufactured by Merck Millipore) arranged on a mesh, spread it evenly, Was dried to prepare a conjugate pad CP.

  Next, the prepared membrane M was bonded to a backing sheet (Adhesives Research, Inc) serving as the support material 111, and then the prepared conjugate pad CP was bonded to the membrane M so as to overlap with the membrane M by several millimeters. Further, on the upstream side in the development direction, a glass fiber (manufactured by Merck Millipore) was bonded as a sample pad SP so as to overlap the conjugate pad CP by several millimeters. On the downstream side of the membrane M, an absorption pad AbP (sample absorption pad, manufactured by Merck Millipore) for receiving excess reagent is arranged to overlap the membrane M several millimeters, and the test strip before applying the cover tape Was made.

  In this verification test, as in the verification test described above, a polyester film (model number PET25 (A) PL Thin 8LK, manufactured by Nippon Biodot Co., Ltd.) was used as the cover tape to be affixed to each test strip. And pasted at each position shown in FIGS. 4A, 4B, 4C, and 4D. Ten test strips were prepared, and each test strip was installed in the case body to prepare multiple test kits. Next, place each test kit horizontally on a flat surface on a desk so that the sample drop opening of the test kit is located on the upper side, and then pass the positive sample with a specified amount of pipette to the sample drop opening of each test kit. The reaction time of each test strip (line detection time) and the reaction state were observed.

  Specifically, the time when the control line and the test line appeared in the determination opening of each test kit was measured as the detection time, and the flow of the solution in each test strip was observed. In addition, 5 minutes after dropping the positive sample, the test kit is destroyed and the sample in the case body is removed to stop the reaction on the test strip, and the line intensity of the line that appears on the test line is determined by the immunochromatography reader. (Measured by Hamamatsu Photonics).

  As a result, in the test strip without the cover tape as a comparative example, the result as shown in FIG. 16 was obtained. In this test strip without cover tape, the average value of 10 lines was 92.3 mAbs, and the average detection time of 10 lines on the test line (indicated as T line in the table) was 47.1 sec. Thus, the average value of the detection time of 10 lines in the control line (denoted as C line in the table) was 38.4 sec. Furthermore, when the flow of the solution on the test strip without the cover tape and the cleanliness of the line were visually confirmed, no particular change of state was noticed.

  Further, in the test strip of the full surface cover tape as another comparative example, the result as shown in FIG. 17 was obtained. From FIG. 17, in the test strip of the full surface cover tape, the line strength was lowered and the detection time of the C line was slightly longer than in the case without the cover tape. Furthermore, on the test strip of the full surface cover tape, when the flow of the solution and the cleanliness of the line were visually confirmed, a streak-like flow such as smoke was generated, and unevenness was confirmed in the flow. Further, the solution from the membrane was not easily removed, and the color of the gold colloidal particles remained on the membrane even after 5 minutes had passed since the dropping of the positive specimen, making it difficult to see the line on the membrane.

  On the other hand, with the test strip 22 of the 5 mm first cover tape of the present invention, the results shown in FIG. 18 were obtained. From FIG. 18, the test strip 22 of the 5 mm first cover tape has an average value of 10 line strengths as compared with the test strip without the cover tape as a comparative example and the test strip with the full cover tape as a comparative example. Was 119.1 mAbs, and it was confirmed that the line strength was higher than that of the comparative example and the line was easy to see. Moreover, the detection time of the T line was also shorter than that of the comparative example, and it was confirmed that the presence or absence of the line could be determined in a short time. Furthermore, with the test strip 22 of this 5mm first cover tape, when the flow of the solution and the cleanliness of the line were visually confirmed, the flow of the solution flowed evenly with no unevenness, and the line was clearly expressed. There was no particular change in condition.

  Further, in the test strip 32 of the 5 mm second cover tape of the present invention, the result as shown in FIG. 19 was obtained. From FIG. 19, the test strip 32 of the 5 mm second cover tape has an average value of 10 line strengths as compared with the test strip without the cover tape as a comparative example and the test strip with the full cover tape as a comparative example. Was 123.5 mAbs, and it was confirmed that the line strength was higher than that of the comparative example and the line was easier to see. Moreover, the detection time of the T line and C line was also shorter than that of the comparative example, and it was confirmed that the presence or absence of the line could be determined in a short time. Furthermore, with the test strip 32 of this 5mm second cover tape, when the flow of the solution and the cleanliness of the line were visually confirmed, the flow of the solution flowed evenly with no unevenness, and the line was clearly expressed. There was no particular change in condition.

  Further, with the test strip 42 of the 9 mm cover tape of the present invention, the results as shown in FIG. 20 were obtained. From FIG. 20, the test strip 42 of 9 mm cover tape also has an average value of 102.2 line strengths as compared with the test strip without the cover tape as a comparative example and the test strip with the full cover tape as a comparative example. It was confirmed that the line strength was higher than that of the comparative example and the line was easier to see than the comparative example. Moreover, the detection time of the T line and C line was also shorter than that of the comparative example, and it was confirmed that the presence or absence of the line could be determined in a short time. Furthermore, in this test strip 42 of the 9mm cover tape, when the flow of the solution and the cleanliness of the line were visually confirmed, the flow of the solution flowed evenly without unevenness, and the line was clearly expressed, There was no particular change in condition.

  Furthermore, with the test strip 52 of the 12 mm cover tape of the present invention, results as shown in FIG. 21 were obtained. From FIG. 21, the 12 mm cover tape test strip 52 has an average value of 147.6 lines compared to the test strip without the cover tape as a comparative example and the test strip with the full cover tape as a comparative example. It was confirmed that the line strength was higher than that of the comparative example and the line was easier to see than the comparative example. Moreover, the detection time of the T line and C line was also shorter than that of the comparative example, and it was confirmed that the presence or absence of the line could be determined in a short time. However, in the test strip 52 of this 12 mm cover tape, when the flow of the solution and the cleanliness of the line were visually confirmed, there was an unevenness in the flow of the solution, and there were also unevenness in the expression state of the line. there were. In addition, some membrane solutions were poorly removed, and even after 5 minutes had passed since the positive specimen was dropped, the color of the colloidal gold particles remained on the membrane and the membrane line was very difficult to see.

  From this, in addition to increasing the line strength and shortening the detection time, in addition to considering how the solution flows on the test strip and maintaining the cleanliness of the line, rather than using 12 mm cover tape, It was confirmed that it is desirable to use 5mm first cover tape, 5mm second cover tape, and 9mm cover tape.

(4-3) About test strip used for detection of influenza virus Next, the verification test was performed about the effectiveness of the cover tape, using influenza A virus and influenza B virus as a positive specimen. In this verification test, a conventional test strip without a cover tape (without a cover tape) is different from the four types of test strips shown in FIGS. 4A, 4B, 4C, and 4D, which have different cover tape application ranges, and conjugates. Not only the gate pad CP but also the test strip (covered with full cover tape) that covers the entire surface of the membrane M is prepared. The detection time until the line was developed in the control line CL was examined. These multiple types of test strips used in this verification test differ only in the cover tape configuration, and support material 111, sample pad, conjugate pad, membrane (here, nitrocellulose membrane is used), absorbent pad The size and material of each were the same.

  The test strip for influenza virus used in this verification test was prepared as follows. First, after preparing a nitrocellulose membrane (HF90 membrane, manufactured by Merck Millipore), a liquid in which a mouse-derived anti-influenza virus type A IgG monoclonal antibody was adjusted to a concentration of 1.0 mg / ml was added at a rate of 1.0 μl / cm. Then, using a positive pressure spray device Biojet xyz3060 (Nippon Biodot Co., Ltd.), a type A test line TL1 was prepared by coating in a line at a predetermined position on the upstream side in the developing direction when viewed from the end of the membrane.

  In addition, a liquid with an anti-influenza virus type B monoclonal antibody concentration of 1.0 mg / ml was viewed from the end of the membrane with a positive pressure spray device Biojet xyz3060 (Japan Biodot) at a rate of 1.0 μl / cm. Then, a B-type test line TL2 was produced by applying in a line shape downstream of the A-type test line TL1 in the development direction. Furthermore, a goat-derived anti-mouse IgG polyclonal antibody was dissolved in PBS, and a solution with a concentration of 1.0 mg / ml was added at a rate of 1.0 μl / ml using a positive pressure spray device Biojet xyz3060 (Japan Biodot Co., Ltd.). A control line (also referred to as a reference line) CL was prepared by coating in a line on the downstream side in the development direction as viewed from the test line TL1 and the B-type test line TL2.

  Then, after each membrane coated with each antibody was placed in a dryer at 40 ° C. and sufficiently dried overnight, the membrane was immersed in a PBS solution containing 1% BSA (bovine serum albumin) and a surfactant for 30 minutes, After blocking the protein non-adsorbed region, the membrane M was dried overnight in an oven to prepare a membrane M used for a verification test.

  Next, a blue colored latex particle-labeled antibody suspension and a red colored latex particle-labeled antibody suspension were prepared as follows. First, in a test tube, a buffer solution (50 mM MES pH 6.0) containing 4% colored latex particles (blue colored latex particles or red colored latex particles) and 0.1 g / mL water-soluble carbodiimide (1-Ethyl -3- (3-dimethylaminopropyl) carbodiimide, hydrochloride, Wako Pure Chemicals) solution, stirred for 40 minutes and reacted, then centrifuged to remove excess water-soluble carbodiimide, and blue colored latex particles Red colored latex particles were obtained. Here, the activated blue colored latex particles were used for influenza A virus, and the activated red colored latex particles were used for influenza B virus.

  Then, mouse-derived anti-influenza virus type A IgG monoclonal antibody (1.0 mg / mL) was added to the activated blue colored latex particles at a ratio of 20: 1 by weight, and stirred overnight at room temperature to give a blue color. After binding the latex particles to the mouse-derived anti-influenza virus type A IgG monoclonal antibody, an unreacted active group was blocked by adding phosphate buffer pH 7.5 containing 1% BSA and reacting overnight. The blue colored latex particle solution thus obtained was centrifuged at 20,000 G for 10 minutes to obtain an influenza A colored latex particle labeled antibody.

  On the other hand, mouse-derived anti-influenza virus type B IgG monoclonal antibody (1.0 mg / mL) was added to the activated red colored latex particles at a weight ratio of 20: 1 and stirred overnight at room temperature. After binding colored latex particles and mouse-derived anti-influenza virus B IgG monoclonal antibody, phosphate buffer pH 7.5 containing 1% BSA was added and reacted overnight to block unreacted active groups. . The red colored latex particle solution thus obtained was centrifuged at 20,000 G for 10 minutes to obtain an influenza B colored latex particle labeled antibody.

  Then, these influenza A colored latex particle labeled antibodies and influenza B colored latex particle labeled antibodies are mixed, and a predetermined amount of 4% sucrose water containing 1% BSA is added and resuspended with a pipette. After preparing the antibody suspension, apply the colored latex particle-labeled antibody suspension at a rate of about 45 μL per 1 cm to a conjugate pad (glass fiber, manufactured by Merck Millipore) 8 mm wide arranged on a net. Spread out uniformly and dry overnight in an oven to make a conjugate pad CP.

  Next, the prepared membrane M was attached to a backing sheet (Adhesives Research, Inc) serving as a support material 111, and then the prepared conjugate pad CP was attached to the membrane M so as to overlap several millimeters. Furthermore, a glass fiber (manufactured by Merck Millipore) was laminated as a sample pad SP on the upstream portion in the development direction so as to overlap the conjugate pad CP by several millimeters. On the downstream side of the membrane M, an absorption pad AbP (sample absorption pad, manufactured by Merck Millipore) for receiving excess reagents is placed so as to overlap the membrane M several millimeters, and the test strip before applying the cover tape Was made.

  As a cover tape to be affixed to each test strip, a polyester film (model number PET25 (A) PL Thin 8LK, manufactured by Nippon Biodot Co., Ltd.) is used, sheared to a predetermined length, and FIGS. 4A, 4B, 4C and 4D. It stuck on each position shown in. In FIG. 4A, FIG. 4B, FIG. 4C and FIG. 4D, only one test line TL1 is shown. However, in this verification test, the test line TL1 is arranged between the test line TL1 and the control line CL. Another test line TL2 (not shown) is also formed to run.

  Incidentally, the configuration of each test strip used in this verification test is the test strip 22 shown in FIG. 4A described above in “(4-1) Test strip using blue colored latex particles” (5 mm first cover tape). ), The test strip 32 (5 mm second cover tape) shown in FIG. 4B, the test strip 42 (9 mm cover tape) shown in FIG. 4C, the test strip 52 (12 mm cover tape) shown in FIG. 4D, and a cover tape as a comparative example Since it is the same as each structure of the test strip (cover cover tape) to which the cover strip 117 covering the entire surface of the test strip, conjugate pad CP and membrane M is attached, the description thereof is omitted here.

  Prepare 10 test strips of 5mm 1st cover tape, 5mm 2nd cover tape, 9mm cover tape, 12mm cover tape, no cover tape, and full cover tape, and install them in the case body. Ten test kits were prepared for each different type. Next, place each test kit horizontally on a flat surface on a desk so that the sample drop opening of the test kit is located on the upper side, and then drop the sample for each test kit with a specified amount (90 μl) pipette. It dropped at the opening and the reaction time (line detection time) and reaction state of each test strip were observed. Two types of positive specimens, influenza A antigen 275 U / mL and influenza B antigen 53 U / mL, were prepared, and a verification test was performed for each positive specimen.

  Specifically, the time at which lines appeared on the control line CL and the test lines TL1 and TL2 at the determination opening of each test kit was measured as the detection time, and the flow of the solution in each test strip was observed. Also, 5 minutes after dropping the positive sample, the test kit is destroyed and the sample in the case body is removed to stop the reaction on the test strip, and the line strength of the lines appearing on the test lines TL1 and TL2 Was measured with an immunochromatographic reader (manufactured by Hamamatsu Photonics).

  As a result, in the test strip without the cover tape as a comparative example, results as shown in FIGS. 22 and 23 were obtained. FIG. 22 shows the results at test line TL1 where influenza A virus can be detected, and FIG. 23 shows the results at test line TL2 where influenza B virus can be detected. As shown in FIG. 22, the average of 10 line intensities of the test line TL1 for detecting influenza A virus is 64.9 mAbs, and the line intensity of the test line TL2 for detecting influenza B virus as shown in FIG. The average value of the 10 was 66.6 mAbs. Also, as shown in FIGS. 22 and 23, the average values of the 10 detection times of each line expression in the test lines TL1 and TL2 were 34.0 sec and 33.7 sec, respectively. When the flow of the solution (positive specimen) on the test strip and the cleanliness of each line that appeared on the test lines TL1 and TL2 were visually confirmed, no particular change in state was noticed.

  With the test strip of the full cover tape as another comparative example, the results as shown in FIGS. 24 and 25 were obtained. FIG. 24 shows the result on the test line TL1 where influenza A virus can be detected, and FIG. 25 shows the result on the test line TL2 where influenza B virus can be detected. As shown in FIG. 24 and FIG. 25, the line strength of the test strip of the full cover tape as another comparative example was lower than that of the test strip without the cover tape. As for the test strip of the full cover tape, when the flow of the solution on the test strip and the cleanliness of each line filed for the test lines TL1 and TL2 were visually confirmed, a streak-like flow occurred. It was confirmed that the flow was uneven. Moreover, in both test lines TL1 and TL2, the solution from the membrane M did not come off well, and the color of each colored latex particle remained on the membrane M even after 5 minutes from the dropping of the positive sample. The line became very difficult to see.

  On the other hand, in the test strip of the 5 mm first cover tape as an example, results as shown in FIGS. 26 and 27 were obtained. FIG. 26 shows the results at test line TL1 where influenza A virus can be detected, and FIG. 27 shows the results at test line TL2 where influenza B virus can be detected. As shown in FIGS. 26 and 27, in the test strip of the 5 mm first cover tape, the average values of the ten line strengths of the test lines TL1 and TL2 are 88.4 mAbs and 78.8 mAbs, respectively. It was confirmed that the line strength was higher than that of the test strip of the full surface cover tape (hereinafter, these are simply referred to as comparative examples), and the lines on the test lines TL1 and TL2 were easier to see.

  In addition, with the test strip of the 5mm first cover tape, the detection time of the lines on the test lines TL1, TL2 and the control line CL was shorter than the comparative example, and it was confirmed that the presence or absence of the line could be judged in a short time . In addition, with the test strip of the 5mm first cover tape, the flow of the solution on the test strip and the cleanliness of the lines that appeared on the test lines TL1 and TL2 were visually confirmed. In addition, the line was clearly expressed, and the state of concern was not particularly observed.

  In the test strip of the 5 mm second cover tape as another example, the results as shown in FIGS. 28 and 29 were obtained. FIG. 24 shows the result on the test line TL1 where influenza A virus can be detected, and FIG. 25 shows the result on the test line TL2 where influenza B virus can be detected. As shown in FIGS. 28 and 29, in the test strip of the 5 mm second cover tape, the average values of the ten line strengths of the test lines TL1 and TL2 are 84.2 mAbs and 77.5 mAbs, respectively, and the line strength is higher than that of the comparative example. It was confirmed that the line on the test lines TL1 and TL2 became easier to see.

  In addition, with the test strip of the 5mm second cover tape, the detection time of the lines on the test lines TL1, TL2 and the control line CL was also shorter than that of the comparative example, and it was confirmed that the presence or absence of the line could be judged in a short time. . Furthermore, with the test strip of the 5mm second cover tape, when the solution flow on the test strip and the cleanliness of the lines that appeared on the test lines TL1 and TL2 were visually checked, the solution flow was uniform and even. In addition, the line was clearly expressed, and the state of concern was not particularly observed.

  In the test strip of the 9 mm cover tape as an example, results as shown in FIGS. 30 and 31 were obtained. FIG. 30 shows the results at test line TL1 where influenza A virus can be detected, and FIG. 31 shows the results at test line TL2 where influenza B virus can be detected. As shown in FIGS. 30 and 31, in the test strip of 9 mm cover tape, the average values of the 10 line strengths of the test lines TL1 and TL2 are 98.9 mAbs and 91.8 mAbs, respectively, and the line strength is higher than that of the comparative example. It was confirmed that the line was easy to see.

  Moreover, in the test strip of 9 mm cover tape, the detection time of the lines in the test lines TL1 and TL2 and the control line CL was also shorter than that in the comparative example, and it was confirmed that the presence or absence of the line could be determined in a short time. Furthermore, with the 9mm cover tape test strip, the flow of the solution on the test strip and the cleanliness of the lines that appeared on the test lines TL1 and TL2 were visually checked. In addition, the line was clearly expressed, and no particular state change was observed.

  In the test strip of the 12 mm cover tape as an example, results as shown in FIGS. 32 and 33 were obtained. FIG. 32 shows the results at test line TL1 where influenza A virus can be detected, and FIG. 33 shows the results at test line TL2 where influenza B virus can be detected. As shown in FIGS. 32 and 33, in the test strip of 12 mm cover tape, the average values of 10 line strengths of the test lines TL1 and TL2 are 74.1 mAbs and 71.5 mAbs, respectively, and the line strength is higher than that of the comparative example. It was confirmed that the line was easy to see. However, with the test strip of 12mm cover tape, the flow of the solution on the test strip and the cleanliness of the lines that appeared on the test lines TL1 and TL2 were visually confirmed. In the expression state, shading unevenness was also confirmed.

  From this, even with test strips for influenza viruses, not only increasing the line strength and shortening the detection time, but also considering how the solution flows on the test strip and maintaining the cleanliness of the line, It was confirmed that it is preferable to use a 5mm first cover tape, a 5mm second cover tape, and a 9mm cover tape rather than using a 12mm cover tape.

1 Inspection kit
101 Case body
2,12,22,32,42,52 Test strip
5,15,25,35 Cover tape (cover material)
105 Sample drop opening
106 Judgment opening
A1, B1 labeled antibody (labeled substance)
A2, B2 antigen capture antibody (capture substance)
SP sample pad
CP conjugate pad
M membrane
AbP absorption pad
TL1, TL2 test line (detection area)
CL control line (operation check area)

Claims (8)

An inspection kit with an inspection strip installed in the case body,
The test strip is
A sample pad to which a specimen extract containing a specimen is applied;
A conjugate pad in which a labeling substance that specifically reacts with the analyte developed from the sample pad by capillary action to form a complex is releasably held in a wet state;
Comprising the complex developed by capillary action, and a membrane having a detection region on which a specifically reacting capture substance is immobilized,
On the upper surface of the case body,
A specimen dropping opening for exposing the sample pad of the test strip to the outside and a determination opening for exposing the membrane of the test strip to the outside are formed,
Between the conjugate pad and the conjugate pad side end of the detection region of the membrane,
A test kit, wherein a cover member is provided so as to cover the entire surface of the conjugate pad from a partial region of the sample pad . An inspection kit with an inspection strip installed in the case body,
The test strip is
A sample pad to which a specimen extract containing a specimen is applied;
A conjugate pad in which a labeling substance that specifically reacts with the analyte developed from the sample pad by capillary action to form a complex is releasably held in a wet state;
Comprising the complex developed by capillary action, and a membrane having a detection region on which a specifically reacting capture substance is immobilized,
On the upper surface of the case body,
A specimen dropping opening for exposing the sample pad of the test strip to the outside and a determination opening for exposing the membrane of the test strip to the outside are formed,
Between the conjugate pad and the conjugate pad side end of the detection region of the membrane,
Said capture inspection kit from some areas you characterized in that the cover member is provided partially or entirely covering the Migihitsuji of the conjugate pad materials the membrane to expose the membrane. An inspection kit with an inspection strip installed in the case body,
The test strip is
A sample pad to which a specimen extract containing a specimen is applied;
A conjugate pad in which a labeling substance that specifically reacts with the analyte developed from the sample pad by capillary action to form a complex is releasably held in a wet state;
Comprising the complex developed by capillary action, and a membrane having a detection region on which a specifically reacting capture substance is immobilized,
On the upper surface of the case body,
A specimen dropping opening for exposing the sample pad of the test strip to the outside and a determination opening for exposing the membrane of the test strip to the outside are formed,
The sample pad side laminated part in which the sample pad and the conjugate pad are laminated, while covering from the membrane side laminated part in which the membrane and the conjugate pad are laminated to a predetermined area of the conjugate pad, inspection kit characterized in that the cover member to expose to the outside without covering the membrane. An inspection kit with an inspection strip installed in the case body,
The test strip is
A sample pad to which a specimen extract containing a specimen is applied;
A conjugate pad in which a labeling substance that specifically reacts with the analyte developed from the sample pad by capillary action to form a complex is releasably held in a wet state;
Comprising the complex developed by capillary action, and a membrane having a detection region on which a specifically reacting capture substance is immobilized,
On the upper surface of the case body,
A specimen dropping opening for exposing the sample pad of the test strip to the outside and a determination opening for exposing the membrane of the test strip to the outside are formed,
The sample pad and the conjugate pad are laminated to the predetermined region of the conjugate pad from the sample pad side laminated portion where the sample pad and the conjugate pad are laminated, while the membrane and the conjugate pad are laminated. inspection kit characterized in that the cover member to expose to the outside without covering the membrane. The cover member is non-permeable any one test kit according one of claims 1-4 to be characterized is a pressure-sensitive adhesive tape made of a synthetic resin member. The membrane is separate from the detection area,
Any one of claims 1-5, characterized in that an indicator capture substance capable of reacting the conjugate pad the deployed labeling substance and specifically from has an operation confirmation area immobilized Inspection kit. It said cover member, a test kit of any one of claims 1-6, characterized in that it is affixed to the test strip. The said cover member is provided in the back surface of the cover body of the said case main body. The inspection kit of any one of Claims 1-6 characterized by the above-mentioned.

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