Nozzle for test pretreatment container, test kit, and sample processing method

Abstract

To achieve the filtration of a specimen sample without lowering sensitivity or while suppressing the lowering of sensitivity.SOLUTION: A nozzle for a test pretreatment container according to an embodiment includes a nozzle main body and a filter, wherein the nozzle main body includes a connection portion connected to a test pretreatment container in which a specimen collected from a patient and a specimen treatment solution that is a treatment solution for the specimen are stored, and a nozzle portion provided in the connection portion and having a communication hole for delivering a liquid stored in the test pretreatment container, the filter is disposed in the nozzle main body and filters the liquid, and a porosity of the filter is 70 to 95%.SELECTED DRAWING: Figure 2

Description

[Technical Field]

[0001] The embodiments disclosed in the present specification and drawings relate to a nozzle for a test pretreatment container, a test kit, and a sample processing method. [Background technology]

[0002] In recent years, rapid test reagents or kits have been developed that utilize antigen-antibody reactions or enzyme reactions to detect or quantify various parameters, such as infection with pathogens such as viruses and bacteria, pregnancy, and urine and blood glucose levels, in a short timeframe of a few minutes to a few tens of minutes. These tests detect or quantify pathogen-constituting proteins, human chorionic gonadotropin (hCG), and glucose in samples. Many rapid test reagents are characterized by their low cost and ease of use, requiring no special equipment. For example, rapid test reagents for pregnancy diagnosis are sold over-the-counter (OTC) at general pharmacies. Furthermore, rapid test reagents for pathogen infection detection, unlike other test reagents, are widely used in general hospitals and clinics, in addition to large hospitals and medical testing centers. Recently, COVID-19 test reagents are also available over-the-counter at general pharmacies. Given this background, rapid test reagents are becoming increasingly important in medical care, as the ability to immediately determine the presence or absence of infection in a patient's sample allows for early treatment before symptoms appear.

[0003] Currently, immunoassays, i.e., immunoagglutination and immunomembrane assays, particularly assays using membranes such as nitrocellulose or filters, are commonly known as simple testing methods. These methods are broadly divided into flow-through and lateral flow membrane assays. Other methods use optical sensors equipped with optical waveguide layers to detect the amount of a target substance in a sample as an optical change. However, these simple testing methods can sometimes result in false positives (positive results when the target substance is not present in the sample) or false negatives (negative results when the target substance is present in the sample) when analyzing samples actually collected from patients. False positive and false negative reactions when measuring pathogen infections can provide incorrect information about the disease, delaying the identification of the cause and potentially leading to inappropriate treatment, exacerbating the condition. Therefore, preventing false positives and false negatives is an extremely important issue given the primary purpose of simple testing methods. The causes of these false positives and false negatives are not fully understood, but one of the possible causes is thought to be non-specific reactive substances contained in the sample, viscosity, and other properties derived from the sample components.

[0004] To solve the above problems, it is effective to filter the sample before performing the assay. Filtration removes substances that cause false positives and false negatives from the sample, preventing these substances from entering the assay and preventing false positives and false negatives. However, when filtering, impurities in the sample can clog the filter pores, preventing the assay from starting. One way to solve this problem is to devise a sample filtration filter configuration. Using a sample filtration filter consisting of multiple filters, ranging from large to small pore sizes or particle retention sizes, can somewhat avoid clogging of the sample during filtration. However, this creates a new problem. First, stacking multiple filters increases the filter's thickness, and when commonly used filters such as cellulose filter paper are used, the filter absorbs the sample, resulting in increased sample loss. To particularly increase sensitivity, it is necessary to minimize the amount of buffer solution and increase the concentration of the analyte in the sample. However, if the filter absorbs a large amount of the analyte, the buffer solution must be increased accordingly, resulting in reduced sensitivity. Furthermore, the more filters are stacked, the greater the likelihood that the target substance will be adsorbed to the filter. While increasing the filtration area is also an option, the buffer volume is limited, making it impossible to increase the area indefinitely and thus not achieving significant results. Therefore, achieving filtration of the sample without or while suppressing a decrease in sensitivity is an important challenge in building a measurement system for a test kit. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Laid-Open No. 2000-088851 Summary of the Invention [Problem to be solved by the invention]

[0006] One of the problems to be solved by the embodiments disclosed in this specification and the drawings is to achieve filtration of a specimen sample without reducing or suppressing a reduction in sensitivity. However, the problems to be solved by the embodiments disclosed in this specification and the drawings are not limited to the above problem. Problems corresponding to the effects of each configuration shown in the embodiments described below can also be considered as other problems. [Means for solving the problem]

[0007] The nozzle for a test pretreatment container according to the embodiment comprises a nozzle body and a filter, the nozzle body having a connection portion connected to a test pretreatment container that contains a sample collected from a patient and a sample treatment liquid that is a treatment liquid for the sample, and a nozzle portion provided in the connection portion and having a communicating hole for discharging the liquid contained in the test pretreatment container, the filter being disposed within the nozzle body and filtering the liquid, and the porosity of the filter being 70 to 95%. [Brief explanation of the drawings]

[0008] [Figure 1] FIG. 1 is a diagram showing the appearance of a test kit according to a first embodiment. [Figure 2] 3A and 3B are diagrams showing an example of the configuration of a nozzle for a pre-inspection treatment container according to the first embodiment. [Figure 3] 3 is a cross-sectional view taken along the vertical direction of the nozzle for the pre-inspection treatment container shown in FIGS. 1 and 2. FIG. [Figure 4] FIG. 2 is a diagram showing an example of a processing procedure for sample processing using the test kit according to the first embodiment. [Figure 5] FIG. 10 is a diagram showing an example of introducing a specimen into a pre-test treatment container. [Figure 6] FIG. 2 is a view showing a state in which a nozzle for a pre-inspection treatment container according to the first embodiment is connected to a pre-inspection treatment container. [Figure 7] 5A and 5B are diagrams for explaining a state in which a mixed liquid is dropped onto a testing device through a pre-test treatment container nozzle according to the first embodiment. [Figure 8]10A and 10B are diagrams showing an example of the configuration of a nozzle for a pre-inspection treatment container according to a second embodiment. [Figure 9] 9 is a cross-sectional view taken along the vertical direction of the nozzle for the pre-inspection treatment container shown in FIG. 8. FIG. [Figure 10] FIG. 10 is a diagram showing the appearance of a test kit according to a third embodiment. [Figure 11] FIG. 11 is a diagram showing an example of a processing procedure for sample processing using a test kit according to a third embodiment. [Figure 12] 10 is a view for explaining a state in which a mixed liquid is dropped into a reservoir via a pre-inspection treatment container nozzle according to the third embodiment. FIG. [Figure 13] FIG. 1 shows the results of evaluation of the risk of specimen-derived clogging for each filter in Comparative Examples 1 to 11 and Example 1. [Figure 14] FIG. 1 shows the results of sensitivity / specificity evaluation. [Figure 15] FIG. 10 shows the results of evaluation of the risk of sample-induced clogging for each filter. [Figure 16] FIG. 10 is a diagram showing the results of evaluation of the relationship between filter thickness and the risk of sample-induced clogging. DETAILED DESCRIPTION OF THE INVENTION

[0009] Hereinafter, embodiments of a nozzle for a test pretreatment container, a test kit, and a sample treatment method will be described with reference to the drawings. In the following description, components having substantially the same functions and configurations will be designated by the same reference numerals, and redundant explanations will be given only when necessary.

[0010] [First embodiment] FIG. 1 is a diagram showing an overview of a test kit according to a first embodiment. Test kit 1 is a kit used when testing a test target substance. The test target substance is not particularly limited, but examples include pathogens such as influenza virus, adenovirus, respiratory syncytial virus (RS) virus, and coronavirus (COVID-19, etc.), as well as parts of these pathogens. As shown in FIG. 1, test kit 1 according to this embodiment includes a nozzle 10 for a test pretreatment container, a test pretreatment container 20, a test device 30, and a specimen collection tool 40.

[0011] The test pretreatment container 20 is configured to be connectable to the test pretreatment container nozzle 10, is deformable by pressure, and is a container that contains a specimen treatment liquid SPL. As an example of the specimen treatment liquid SPL, a buffer solution containing a surfactant is used. The specimen is suspended in this specimen treatment liquid SPL. The test pretreatment container 20 is made of a material that can be deformed by manual pressure. The material of the test pretreatment container 20 is, for example, a plastic material such as a thermoplastic resin. Examples of this thermoplastic resin include, but are not limited to, polypropylene, polyethylene, polyvinyl chloride, polyethylene terephthalate, and polystyrene. Polyethylene or polypropylene is preferred due to its excellent chemical resistance and versatility.

[0012] The test device 30 has a light-transmitting substrate (hereinafter referred to as a light-transmitting substrate) on which a first substance that specifically binds to a test target substance is immobilized. The test device 30 is provided with a drip hole that connects to a reaction chamber provided inside. The test device 30 is attached to an optical measurement device (not shown), and the test target substance is measured by the optical measurement device. This test device 30 is also called a test cartridge.

[0013] The specimen collection tool 40 is an instrument for collecting a specimen from a patient. The specimen collection tool 40 may be, for example, a cotton swab, a platinum loop, a dropper, or a spoon-shaped tool. The specimen is preferably a throat swab, a nasal swab, a nasopharyngeal swab, a nasal aspirate, or a nasal wash collected from a human or other animal, but may also be alveolar lavage fluid, blood, a fecal suspension, or a rectal swab. In particular, when the subject is a human and the specimen is a throat swab, a nasal swab, a nasal aspirate, a nasal wash, a fecal suspension, or a rectal swab, a cotton swab is used as the specimen collection tool 40. As shown in FIG. 1, the cotton swab has a specimen collection portion 41 formed from a cotton ball with cotton wrapped around a head at one end of a shaft 42. However, the configuration of the cotton swab is not limited to this. That is, the configuration of the cotton swab is arbitrary, and it may be, for example, a brush-like cotton swab in which natural or artificial fibers are arranged in a brush shape and which uses capillary action to collect liquid.

[0014] The nozzle 10 for the pre-inspection treatment container is a nozzle connected to the pre-inspection treatment container 20. FIG. 2 is a diagram showing an example of the configuration of the nozzle 10 for the pre-inspection treatment container according to the first embodiment. FIG. 3 is a cross-sectional view along the vertical direction of the nozzle 10 for the pre-inspection treatment container shown in FIGS. 1 and 2. As shown in FIGS. 2 and 3, the nozzle 10 for the pre-inspection treatment container according to this embodiment is configured to include a nozzle main body 11, a filter 12, and a fall-off prevention member 13.

[0015] As shown in Fig. 3, the nozzle body 11 has a connection portion 111 and a nozzle portion 113. In this embodiment, the connection portion 111 and the nozzle portion 113 are formed from an elastic synthetic resin such as low-density polyethylene. Note that in this embodiment, the nozzle portion 113 is configured as an integral part of the connection portion 111, but these may also be formed separately and then joined together. Alternatively, the nozzle portion 113 may be configured so that the user can attach the nozzle portion 113 to the connection portion 111 later.

[0016] The connecting part 111 is connected to the pre-test treatment container 20, which contains a sample collected from a patient and a sample treatment liquid SPL, which is a treatment liquid for the sample. The connecting part 111 according to this embodiment is connected to the pre-test treatment container 20 by being fastened to the pre-test treatment container 20 by screwing. For this reason, the connecting part 111 is generally cylindrical, and a screw groove 1111 is formed on the inner side of the connecting part 111. The formed screw groove 1111 may be one or more. In the example shown in FIG. 2 , a knurling 1112 is provided on the outer side of the connecting part 111. This knurling 1112 is intended to prevent the user's finger from slipping when attaching the connecting part 111 to the pre-test treatment container 20. Note that, in this embodiment, the knurling 1112 is provided on the outer periphery of the connecting part 111 to prevent the user's finger from slipping, but the configuration for preventing the user's finger from slipping is not limited to this. That is, the configuration for preventing the user's finger from slipping is arbitrary, and for example, one or more convex portions may be formed extending from above to below on the side surface of connecting portion 111 so as to have a convex shape in the outer periphery direction. Also, in this embodiment, a configuration for preventing the finger from slipping is provided on the outer periphery of connecting portion 111, but the configuration for preventing the finger from slipping is not necessarily provided.

[0017] The nozzle part 113 is provided in the connection part 111, and has a communication hole 1131 formed therein for delivering the liquid contained in the pre-test treatment container 20. Specifically, as shown in FIGS. 2 and 3, the nozzle part 113 is roughly shaped like an elongated cylinder with the communication hole 1131 formed in its center. The outer and inner diameters of the cylinder of the nozzle part 113 are smaller than the outer and inner diameters of the connection part 111, respectively, and the length of the cylinder of the nozzle part 113 is longer than the length (height) of the connection part 111. Furthermore, the nozzle part 113 is provided concentrically with the connection part 111 on an upper surface 1113 of the connection part 111.

[0018] The communicating hole 1131 forming the cylindrical inner diameter of the nozzle portion 113 is connected to the cavity forming the cylindrical inner diameter of the connection portion 111, and is configured so that the liquid from the pre-test treatment container 20 flows smoothly from the cavity of the cylindrical connection portion 111 to the cavity of the cylindrical nozzle portion 113.

[0019] In particular, as shown in Fig. 3, a cavity 1114 is formed in the upper portion of the connection part 111. As shown in Fig. 3, in this embodiment, the filter 12 and the fall-off prevention member 13 are disposed in this cavity 1114. Furthermore, a cavity 1115 is formed in the lower portion of the connection part 111. In this embodiment, the upper end portion of the pre-test treatment container 20 is screwed into this cavity 1115.

[0020] 3, the nozzle portion 113 is formed with a long, thin, linear communication hole 1131. That is, the liquid from the pre-test treatment container 20 that has passed through the filter 12 arranged in the cavity 1114 passes through the communication hole 1131 that communicates with the cavity 1114 and is discharged from the tip of the nozzle portion 113. At this time, the pre-test treatment container nozzle 10 and the pre-test treatment container 20 are used by the user upside down compared to the drawings, with the tip of the nozzle portion 113 facing downward and dripping the liquid into the drip port of the testing device 30. The drip amount of this pre-test treatment container nozzle 10 is set so that, for example, six drops is approximately 160 mg.

[0021] The fall-off prevention member 13 is a member for preventing the filter 12 from falling off. The fall-off prevention member 13 is formed in a cylindrical shape and is provided with a passage hole 131 for passing a liquid. In the example shown in Fig. 3, the fall-off prevention member 13 fits into the cavity 1114, thereby disposing the filter 12 in the cavity 1114 and preventing the filter 12 from falling off from the cavity 1114.

[0022] The filter 12 is disposed within the nozzle body 11 and filters the liquid. Specifically, the filter 12 filters a mixture of a specimen and a specimen treatment liquid, which is a treatment liquid for the specimen. The filter 12 has, for example, a sponge-like structure with pores arranged in a mesh pattern. The outer diameter of the filter 12 is, for example, approximately 0.7 cm. In this embodiment, the filter 12 holds a reagent containing a substance capable of binding to the analyte in the specimen. Therefore, when the mixture of the specimen and the specimen treatment liquid passes through the filter 12, the filter 12 mixes the liquid with the reagent contained in the filter 12 and delivers the mixed liquid to the communicating hole 1131 of the nozzle portion 113.

[0023] The porosity of filter 12 is preferably 70% to 95%, and more preferably 74% to 91%. When the porosity of filter 12 is 70% or more, the risk of clogging of the filter due to the sample can be reduced.

[0024] The pore size of filter 12 is preferably 25 to 600 μm. When filter 12 has a pore size of 25 to 600 μm, the risk of clogging due to the sample can be reduced.

[0025] Furthermore, when filter 12 is pressurized at a pressure of 0.2 MPa, the rate of change in film thickness of filter 12 before and after pressurization is preferably 15% or more, and more preferably 20% or more. When the rate of change in film thickness of filter 12 before and after pressurization is 20% or more, processing is easy and there is a high degree of freedom in pressurizing filter 12 into fine flow paths.

[0026] Furthermore, filter 12 is a so-called depth filter that captures impurities in the specimen not only on the surface but also inside filter 12. For this reason, filter 12 needs to have a certain thickness. For example, the filter thickness is preferably 1 to 5 mm, and more preferably 1.5 to 5 mm. In particular, a filter thickness of 1.5 to 5 mm makes it less likely for clogging due to the specimen to occur, and allows a constant amount of liquid to be dripped.

[0027] Furthermore, the material of the filter 12 is preferably, for example, polyvinyl alcohol, urethane, polyethylene, ethylene / polymer, or polyurethane. However, the material of the filter 12 is not limited to these. That is, the material of the filter 12 is arbitrary. The material of the filter 12 may be other than polyvinyl alcohol, urethane, polyethylene, ethylene / polymer, or polyurethane, and further, a plurality of materials selected from these materials and other materials may be used in any combination.

[0028] Next, a procedure for sample processing using the test kit 1 according to the first embodiment will be described. FIG. 4 is a diagram showing an example of a procedure for sample processing using the test kit 1 according to the first embodiment. As shown in FIG. 4, first, a user collects a sample (step S11). Specifically, the user collects a sample from a patient using a sample collection tool 40. More specifically, for example, if the sample is a nasal swab, the user scrapes the nasopharynx with a sterile cotton swab, which is an example of the sample collection tool 40, to collect the nasal swab.

[0029] 4, the user introduces the specimen into the pre-test treatment container 20 (step S13). Specifically, the user introduces the specimen collected by the specimen collecting tool 40 in step S11 into the pre-test treatment container 20. More specifically, for example, if the specimen is a nasal swab, the user introduces the specimen by immersing the specimen collecting tool 40 in the specimen treatment liquid SPL contained in the pre-test treatment container 20.

[0030] 5 is a diagram showing an example of introducing a specimen into the pre-test treatment container 20. As shown in FIG. 5(a), a user immerses the specimen collecting portion 41 of a sterile cotton swab, which is a specimen collecting tool 40 that has collected a nasal swab, in the specimen treatment liquid SPL contained in the pre-test treatment container 20. Then, as shown in FIG. 5(b), the user applies pressure to the pre-test treatment container 20 using their fingers F to apply pressure to the specimen collecting portion 41 in the pre-test treatment container 20, while moving the sterile cotton swab up and down in the direction D1 and rotating it in the direction R1 to squeeze out the specimen from the specimen collecting portion 41 and introduce the specimen into the pre-test treatment container 20.

[0031] Next, as shown in Fig. 4, the user connects the nozzle 10 for the pre-inspection treatment container to the pre-inspection treatment container 20 (step S15). Fig. 6 is a diagram showing a state in which the nozzle 10 for the pre-inspection treatment container according to the first embodiment is connected to the pre-inspection treatment container 20. As shown in Fig. 6, the user connects the nozzle 10 for the pre-inspection treatment container to the pre-inspection treatment container 20 by screwing the nozzle 10 for the pre-inspection treatment container to the upper end of the pre-inspection treatment container 20.

[0032] 4, the user drips the liquid contained in the pre-test treatment container 20 onto the testing device 30 (step S17). Specifically, the user drips the mixed liquid of the specimen contained in the pre-test treatment container 20 and the specimen treatment liquid SPL onto the testing device 30. More specifically, the user applies pressure to the pre-test treatment container 20 using his or her finger F to filter the mixed liquid through the filter 12 of the pre-test treatment container nozzle 10, and mixes the mixed liquid with the reagent held in the filter 12, and drips the mixed liquid onto the testing device 30 as shown in FIG.

[0033] By executing this step S17, the sample processing according to this embodiment is completed. After executing this step S17, the user sets the inspection device 30 in an optical measurement apparatus (not shown) and starts optical measurement.

[0034] As described above, the nozzle 10 for a test pretreatment container according to the first embodiment comprises a nozzle body 11 and a filter 12, and the filter 12 placed within the nozzle body 11 is configured to have a porosity of 70 to 95%, thereby enabling filtration of the specimen sample without reducing sensitivity or while suppressing a reduction in sensitivity.

[0035] Second Embodiment In the nozzle 10 for a test pretreatment container according to the first embodiment described above, the filter 12 holds a reagent, but this is not limited to this. The nozzle 10 for a test pretreatment container according to the second embodiment may also include a reagent holding section that holds a reagent. Below, the case where this modified example is applied to the first embodiment will be referred to as the second embodiment, and differences from the first embodiment will be described.

[0036] Fig. 8 is a diagram showing an example of the configuration of a nozzle for a test pretreatment container according to the second embodiment, and is a diagram corresponding to Fig. 2. Fig. 9 is a cross-sectional view along the vertical direction of the nozzle for a test pretreatment container shown in Fig. 8, and is a diagram corresponding to Fig. 3. As shown in Figs. 8 and 9, a nozzle 10a for a test pretreatment container according to this embodiment is configured to include a nozzle body 11, a filter 12a, a fall-off prevention member 13a, and a reagent holding portion 14. Note that the configuration of the nozzle body 11 is the same as that of the first embodiment, and therefore description thereof will be omitted.

[0037] The filter 12a according to the second embodiment differs from the first embodiment in that it does not hold a reagent. The fall-off prevention member 13a according to the second embodiment is a member that prevents the filter 12a and the reagent holding portion 14 from falling off. The other configurations of the filter 12a and the fall-off prevention member 13a are the same as those of the filter 12 and the fall-off prevention member 13 according to the first embodiment described above, and therefore description thereof will be omitted.

[0038] The reagent holding unit 14 is a member that holds a reagent containing a substance that can bind to the target substance in the sample. This reagent holding unit 14 is also called a pad. As shown in FIG. 9 , the reagent holding unit 14, the filter 12a, and the fall-off prevention member 13 are arranged in this order from the tip side of the nozzle unit 113 within the cavity 1114. Therefore, the liquid filtered by the filter 12a is delivered to the reagent holding unit 14, and the reagent held in the reagent holding unit 14 and the liquid filtered by the filter 12a are mixed and delivered to the communication hole 1131 of the nozzle unit 113.

[0039] The reagent holder 14 retains the reagent by impregnating it with the reagent and drying it. To allow the reagent to be impregnated, the reagent holder 14 is formed, for example, of a porous structure having a plurality of holes formed therein. The reagent holder 14 according to this embodiment is made of, for example, polyester, and has a thickness of 0.4 mm and a diameter of 0.7 cm. While the material of the reagent holder 14 according to this embodiment is polyester, the material of the reagent holder 14 is not limited to this. The material of the reagent holder 14 is arbitrary and may be, for example, polypropylene, polyethylene, polystyrene, polymethyl methacrylate, alumina, zirconia, or silicon. Although the thickness of the reagent holder according to this embodiment is 0.4 mm, the thickness is not limited to this. The thickness of the reagent holder 14 is arbitrary and may be less than 0.4 mm or greater than 0.4 mm.

[0040] Third Embodiment In the test kits 1 according to the first and second embodiments described above, a liquid containing a specimen, a specimen treatment liquid, and a reagent is dripped onto the test device 30. In the third embodiment, a liquid containing a specimen, a specimen treatment liquid, and a reagent is dripped onto a storage container separate from the test device. Below, the third embodiment will be described as a case where this modified example is applied to the first embodiment, and the differences from the first embodiment will be explained.

[0041] Figure 10 is a diagram showing the appearance of a test kit according to a third embodiment, and is a diagram corresponding to Figure 1. As shown in Figure 10, the test kit 1a according to the third embodiment includes a pre-test treatment container nozzle 10, a pre-test treatment container 20, a test device 30, a specimen collecting tool 40, a storage container 50, and a dropper 60. Note that the configurations of the pre-test treatment container nozzle 10, the pre-test treatment container 20, the test device 30, and the specimen collecting tool 40 are the same as those in Figure 1, and therefore description thereof will be omitted.

[0042] The storage container 50 is a container for storing a liquid containing a specimen, a specimen treatment liquid, and a reagent. The container used for the storage container 50 is not particularly limited as long as it is capable of storing a liquid. However, a container with a lid is preferable to avoid contamination by outside air, etc. In the example shown in FIG. 10, the storage container 50 has a tapered shape that narrows from the opening to the tip. An example of a container that meets these conditions is a microtube.

[0043] The dropper 60 is a tool for injecting the liquid stored in the storage container 50 and discharging it into the testing device 30. The dropper 60 is not an essential component of the test kit 1a. For example, if it is assumed that the operator will use another dropper that they have separately, there is no need to include the dropper 60 in the test kit 1a.

[0044] The procedure for sample processing using the test kit 1a according to the third embodiment will be described with reference to Figure 11. Figure 11 is a diagram showing an example of the procedure for sample processing using the test kit 1a according to the third embodiment, and corresponds to Figure 4. Note that the processing from step S11 to step S15 is the same as in Figure 4, and therefore description thereof will be omitted.

[0045] 11, the user drips the liquid contained in the test pretreatment container 20 into the storage container 50 (step S17a). Specifically, the user drips the mixture of the specimen, specimen treatment liquid SPL, and reagent contained in the test pretreatment container 20 into the storage container 50. More specifically, the user pressurizes the test pretreatment container 20 with his / her finger F, thereby filtering the mixture through the filter 12 of the test pretreatment container nozzle 10, and mixing the mixture of the reagent, specimen, and specimen treatment liquid SPL held in the filter 12, and dripping it into the storage container 50 as shown in FIG.

[0046] By executing step S17a, the sample processing according to this embodiment is completed. After executing step S17a, the user leaves the dropped liquid in the storage container 50 until a predetermined time has passed after dropping it, and after the predetermined time has passed, aspirates the liquid in the storage container 50 with the dropper 60 and drops the aspirated liquid onto the testing device 30 set in the optical measurement apparatus, thereby starting optical measurement.

[0047] [Modification] In the first to third embodiments described above, the nozzle 10 for the pre-inspection treatment container and the pre-inspection treatment container 20 are connected by being fastened by screwing, but the method of connecting the nozzle 10 for the pre-inspection treatment container and the pre-inspection treatment container 20 is not limited to this. That is, the method of connecting the nozzle 10 for the pre-inspection treatment container and the pre-inspection treatment container 20 is arbitrary, and the nozzle 10 for the pre-inspection treatment container and the pre-inspection treatment container 20 may be connected by fitting or by adhesion.

[0048] Furthermore, although the nozzle 10 for a pre-test treatment container according to the first to third embodiments is provided with the fall-off prevention members 13, 13a, it is not necessary to provide the fall-off prevention members 13, 13a. In this case, the filter 12, 12a and the reagent holding part 14 of the nozzle 10 for a pre-test treatment container according to the first to third embodiments may be attached to the nozzle body 11 by adhesive or the like to prevent them from falling off from the nozzle body 11. [Example]

[0049] The present invention will be specifically described below with reference to examples, but the examples described below do not limit the present invention in any way.

[0050] [Test Example 1] Evaluation of the risk of clogging due to specimens 1. Filter Preparation As shown in Table 1, the filters according to Comparative Examples 1 to 11 and Example 1 were packed and prepared. [Table 1]

[0051] 2. Preparation of Sample Solution A brush-shaped swab for specimen collection, which is a specimen collection tool 40, is inserted into a person's nasal cavity to collect a mucosal scraping specimen. The weight of the brush-shaped swab that collected the nasal mucosal specimen (nasopharyngeal swab) is measured, and the collected specimen volume is recorded by subtracting the weight before collection. 550 μL of specimen treatment liquid SPL is dispensed into a pre-test treatment container 20 that can be connected to a pre-test treatment container nozzle 10 equipped with a filter, and the cotton ball portion (specimen collection portion 41) of the brush-shaped swab that collected the nasal mucosal specimen is immersed in the liquid. The cotton ball portion of the brush-shaped swab is pinched through the tube, and the brush-shaped swab is pulled up while squeezing the cotton ball approximately 10 times. Hereinafter, the resulting mixture of the nasal mucosal specimen and specimen treatment liquid is referred to as the sample liquid.

[0052] 3. Evaluation of the amount of sample liquid dropped A test pretreatment container nozzle 10 equipped with a filter is connected to the test pretreatment container 20 containing the sample liquid prepared in 2. The sample liquid is dripped into a microtube whose tare weight has been measured in advance by applying pressure with a finger. Six drops are dripped, and if dripping six drops is difficult due to a clogged filter, squeeze out as much as possible. The amount of dripped liquid for six drops is approximately 160 mg. After dripping is complete, the microtube is weighed and the amount of dripped sample liquid is evaluated by subtracting the tare weight. The evaluation criterion used was whether or not a minimum amount of liquid capable of being tested using test kits 1 and 1a was dripped.

[0053] 4. Results of assessment of the risk of clogging caused by samples FIG. 13 shows the evaluation results for the risk of specimen-derived clogging for each filter in Comparative Examples 1 to 11 and Example 1. As shown in FIG. 13, the horizontal axis represents the amount of nasal mucosa scraping specimen (specimen addition amount) (mg) added, and the vertical axis represents the amount of dripped liquid (mg). The dotted line indicates the minimum testable liquid volume, which is the minimum amount of sample liquid required to perform a test using the test kits 1 and 1a. As a result, among the filters in Comparative Examples 1 to 11, the majority of filters became unable to drip before six drops due to specimen-derived clogging, and did not meet the minimum testable liquid volume. Among the filters in Comparative Examples 1 to 11, the filters in Comparative Example 2 and Comparative Example 10 exceeded the minimum testable liquid volume, but the margin between them and the minimum testable liquid volume for the test kits 1 and 1a was small, and therefore not sufficient. On the other hand, the filter in Example 1 was able to drip a constant amount of sample liquid regardless of the amount of specimen added, and it can be said that the risk of specimen-derived clogging is low.

[0054] [Test Example 2] SARS coronavirus detection sensitivity / specificity evaluation 1. Filter Preparation In this test example, a nozzle 10 for a test pretreatment container was fabricated using the filter according to Example 1 or the filter according to Comparative Example 1. In this test example, the nozzle 10 for a test pretreatment container was equipped with each filter. In this test example, the nozzle 10 for a test pretreatment container was equipped with a reagent holding section 14 on the nozzle side of the filter within the nozzle body. This reagent holding section 14 held a reagent for detecting SARS coronavirus.

[0055] 2. Preparation of Sample Solution A sample treatment solution containing no antigen protein was prepared as a negative sample, and a sample treatment solution containing purified SARS coronavirus antigen protein was prepared as a positive sample. Additionally, nasal mucosa samples were collected from SARS coronavirus-negative subjects using a brush-shaped cotton swab, and the negative and positive samples were added to prepare "negative nasal mucosa samples" and "positive nasal mucosa samples," respectively. Two brush-shaped cotton swabs worth of nasal mucosa samples were collected from the subjects and distributed between the filters of Example 1 and Comparative Example 1. For the sample solution used with the test pretreatment container nozzle 10 using the filter of Comparative Example 1, the amount of nasal mucosa sample added was adjusted to a level that would not cause clogging.

[0056] 3.Evaluation of the impact on sensitivity / specificity Each sample liquid prepared in 2. of Test Example 2 was dropped onto the test device 30 through the nozzle 10 for the test pretreatment container prepared in 1., and the sensitivity / specificity was evaluated. In this test example, the amount of the substance to be detected in the solution was detected as an optical change by using an optical sensor equipped with an optical waveguide layer, but this is not limited thereto, and detection methods using various immunoassays such as immune agglutination and immunomembrane assay methods may also be used.

[0057] 4. Results of sensitivity / specificity evaluation FIG. 14 shows the results of the sensitivity / specificity evaluation. As shown in FIG. 14, when a nasal mucosa sample was not used, there were no problems with sensitivity or specificity for either the filter of Example 1 or the filter of Comparative Example 1. However, when nasal mucosa was added, the filter of Comparative Example 1 exhibited a decrease in sensitivity in the presence of SARS coronavirus antigen protein. For example, if a sensitivity value of 15% was used as the threshold for judgment, the specificity would also decrease (in this case, a false negative). On the other hand, the filter of Example 1 did not exhibit a significant decrease in sensitivity even when nasal mucosa was added, indicating that there was no effect on sensitivity / specificity. Therefore, based on the results of Test Examples 1 and 2, the filter of Example 1 can be used to filter the specimen without or while suppressing a decrease in sensitivity. Furthermore, the filter of Example 1 can remove substances that cause false positives and false negatives from the specimen, thereby preventing false positives and false negatives.

[0058] [Test Example 3] Evaluation of the risk of clogging due to specimens in high porosity filters In the first to third embodiments described above, the filter must have a high porosity to prevent clogging when filtering a sample liquid containing a nasal mucosa specimen and to enable removal of substances that cause false positives and false negatives in the sample. Here, high porosity means a porosity of 70% or more.

[0059] 1. Evaluation of the risk of clogging due to specimens in high-porosity filters The filters shown in Table 2, which have the same porosity as the filter of Example 1, were subjected to an evaluation of the risk of clogging due to the sample in accordance with the method of Test Example 1. [Table 2]

[0060] 2. Results of assessment of the risk of clogging caused by specimens FIG. 15 shows the evaluation results for the risk of specimen-derived clogging for each filter. In FIG. 15, the horizontal axis represents the amount of nasal mucosa scraping specimen added (amount of specimen added), and the vertical axis represents the amount of dripped liquid. Also in FIG. 15, the dotted line indicates the minimum testable liquid volume, which is the minimum amount of sample liquid required to perform a test with the test kit. As shown in FIG. 15, at the specimen addition volume where specimen-derived clogging occurred in the filter of Comparative Example 1 (the amount of dripped liquid was less than or close to the minimum testable liquid volume), specimen-derived clogging did not occur in the filters of Examples 1 to 6, and a constant amount of sample liquid could be dripped, indicating a low risk of specimen-derived clogging.

[0061] [Test Example 4] Analysis of specimen-derived clogging risk and porosity The relationship between the risk of sample-derived filter clogging and porosity is analyzed from the filter of Comparative Example 1, in which sample-derived filter clogging occurred, and the filters of Examples 1 to 6. Since the porosity of the filter of Comparative Example 1, in which sample-derived filter clogging occurred, has not been made public, the porosity of the filter of Comparative Example 1 is calculated for comparative evaluation with the filters of Examples 1 to 6.

[0062] 1. Calculation of porosity By placing a heavy object on the filter, a uniform pressure (17.6 MPa) is applied in the same direction as the filtration direction of the sample liquid, crushing the filter. At this time, the filter membrane thickness before and after pressure application is measured with a micrometer. The rate of change in membrane thickness before and after pressure application is calculated from the measured values. Since the rate of change in membrane thickness before and after pressure application depends on the void ratio (porosity) inside the filter, the rate of change in membrane thickness before and after pressure application is considered to be approximately equal to the porosity. Three filters were measured for each filter, and the average value was calculated.

[0063] 2.Porosity evaluation results Table 3 shows the calculated porosities of the filter according to Comparative Example 1 and the filters according to Examples 1 to 6. For the filters according to Examples 1 to 6, whose porosities are published as catalog values, the porosities calculated in this test example were close to the catalog values. Based on this, the porosity calculation method used in this test example was determined to be appropriate. Furthermore, the porosity of the filter according to Comparative Example 1, in which sample-induced filter clogging occurred, was 59.8%, while the minimum porosity of the filters according to Examples 1 to 6, in which the risk of sample-induced filter clogging was low, was 75.4%. Therefore, a porosity of 70% or more is desirable to reduce the risk of sample-induced filter clogging. In other words, the results of Test Examples 1 to 4 indicate that a filter porosity of 70% or more allows for filtration of the sample without or while suppressing a decrease in sensitivity. [Table 3]

[0064] [Test Example 5] Analysis of high porosity and flexibility In order to maintain the filter structure for filtering sample liquid in a high-porosity filter, it is necessary for the filter to be flexible enough to deform flexibly in response to the sample inflow pressure. For this reason, we assumed that the pressure exerted on the filter by the liquid and air when filtering the sample liquid was approximately 0.2 MPa, and evaluated the flexibility of the filters according to Examples 1 to 6 from the rate of change in film thickness before and after filtration.

[0065] 1. Calculating flexibility The film thickness of the filter to be evaluated is measured in advance using a micrometer. To evaluate flexibility, various filters are immersed in the sample processing liquid SPL for 5 minutes, simulating the time when the sample liquid is filtered. After that, a digital force gauge (manufacturer: IMADA, model number: DST-20N) is used to apply a pressure of 0.2 MPa in the same direction as the filtration direction of the sample liquid. The film thickness after pressure is measured using a micrometer, and the flexibility during filtration of the sample liquid is evaluated from the rate of change in film thickness before and after pressure. Three filters of each type were measured, and the average value was calculated.

[0066] 2. Flexibility evaluation results Table 4 shows the calculation results for the flexibility of the filters according to Examples 1 to 6. For an assumed pressure of 0.2 MPa when filtering the sample liquid, the filters according to Examples 1 to 6 had a pressure-induced thickness change rate of 22.5% to 78.3%. From this, it is desirable that a high-porosity filter with a porosity of 70% or more has a flexibility such that the thickness change rate for an assumed pressure of 0.2 MPa when filtering the sample liquid is 20% or more. Furthermore, a flexibility of 20% or more can be said to facilitate processing and provide a high degree of freedom when pressing the filter into a fine flow path. [Table 4]

[0067] [Test Example 6] Analysis of sample-derived clogging risk and filter thickness For the filters according to Comparative Example 1 and Example 1, the relationship between the filter membrane thickness and the risk of clogging due to the specimen was evaluated. 1. Filter Preparation The filter according to Comparative Example 1 and the filter according to Example 1, in which clogging due to the specimen had occurred, were filled to a film thickness of 1 to 5 mm, and filters were fabricated by the method according to 1. of Test Example 1.

[0068] 2. Risk assessment of sample-derived clogging The test was carried out in accordance with Test Example 1.

[0069] 3. Results of assessment of the risk of clogging caused by samples FIG. 16 shows the evaluation results regarding the relationship between filter film thickness and the risk of sample-induced clogging. In FIG. 16(a), the horizontal axis represents the amount of nasal mucosa scraping specimen added (sample addition amount) and the vertical axis represents the amount of dripped liquid. In the filter of Comparative Example 1, sample-induced clogging occurred when approximately 90 mg of specimen was added, while in the filter of Example 1, a greater amount of specimen was added. In FIG. 16(b), the horizontal axis represents filter film thickness and the vertical axis represents the amount of dripped liquid. In the filter of Example 1, sample-induced clogging did not occur within the film thickness range of 1.5 to 5 mm, even when an amount of specimen added exceeding the amount at which sample-induced clogging occurred in the filter of Comparative Example 1, and a constant amount of sample liquid could be dripped.

[0070] According to at least one of the embodiments described above, filtration of a specimen sample can be achieved without reducing sensitivity or while suppressing reduction in sensitivity.

[0071] Although several embodiments have been described above, these embodiments are presented only as examples and are not intended to limit the scope of the invention. The novel apparatus and method described herein may be embodied in various other forms. Furthermore, various omissions, substitutions, and modifications may be made to the forms of the apparatus and method described herein without departing from the spirit of the invention. The appended claims and their equivalents are intended to cover such forms and modifications that fall within the scope and spirit of the invention. [Explanation of symbols]

[0072] 1, 1a... test kit, 10... nozzle for test pretreatment container, 11... nozzle body, 12, 12a... filter, 13, 13a... fall-off prevention member, 14... reagent holding part, 20... test pretreatment container, 30... test device, 40... specimen collection tool, 41... specimen collection part, 42... shaft part, 50... storage container, 60... dropper, SPL... specimen treatment liquid

Claims

1. a nozzle body having a connection part connected to a pre-test treatment container that contains a specimen collected from a patient and a specimen treatment liquid that is a treatment liquid for the specimen, and a nozzle part provided at the connection part and having a communication hole for delivering the liquid contained in the pre-test treatment container; a filter disposed within the nozzle body for filtering the liquid; A nozzle for a pre-inspection treatment container, wherein the porosity of the filter is 70 to 95%.

2. 2. The nozzle for a pre-inspection treatment container according to claim 1, wherein the pore diameter of the filter is 25 to 600 μm.

3. 2. The nozzle for a pre-inspection treatment container according to claim 1, wherein when the filter is pressurized at a pressure of 0.2 MPa, the rate of change in film thickness of the filter before and after pressurization is 15% or more.

4. 3. The nozzle for a pre-inspection treatment container according to claim 2, wherein when the filter is pressurized at a pressure of 0.2 MPa, the rate of change in film thickness of the filter before and after pressurization is 15% or more.

5. 2. The nozzle for a pre-inspection treatment container according to claim 1, wherein the filter has a film thickness of 1 to 5 mm.

6. The nozzle for a test pretreatment container according to claim 1 , wherein the filter captures impurities in the specimen not only on the surface of the filter but also inside the filter.

7. 2. The nozzle for a pre-inspection treatment container according to claim 1, wherein the filter has a sponge-like structure in which pores are spread out in a mesh-like structure.

8. 2. The nozzle for a test pretreatment container of claim 1, wherein the sample is a throat swab, a nasal swab, a nasopharyngeal swab, a nasal aspirate, a nasal wash, an alveolar lavage fluid, blood, a fecal suspension, or a rectal swab collected from a human or other animal.

9. The nozzle for a test pretreatment container according to claim 1 , wherein the filter holds a reagent containing a substance capable of binding to the target substance in the specimen.

10. The nozzle for a test pretreatment container according to claim 1 , further comprising a reagent holding section for holding a reagent containing a substance capable of binding to the target substance in the specimen.

11. The nozzle for a pre-test treatment container according to claim 10 , wherein the reagent holding portion is formed of a porous structure having a plurality of holes formed therein.

12. The nozzle for a pre-inspection treatment container according to claim 1 , further comprising a fall-off prevention member for preventing the filter from falling off.

13. The nozzle for a pre-inspection treatment container according to claim 1 , wherein the connecting portion is connected to the pre-inspection treatment container by being fastened to the pre-inspection treatment container by screwing.

14. A nozzle for a pre-inspection treatment container according to any one of claims 1 to 13; a test pretreatment container configured to be connectable to the test pretreatment container nozzle, deformable by pressure, and containing a specimen treatment liquid that is a treatment liquid for the specimen; a test device for detecting or quantifying a target substance in the sample; a specimen collection tool for collecting the specimen from the patient; A test kit comprising:

15. A nozzle for a pre-inspection treatment container according to any one of claims 1 to 13; a test pretreatment container configured to be connectable to the test pretreatment container nozzle, deformable by pressure, and containing a specimen treatment liquid that is a treatment liquid for the specimen; a specimen collection tool for collecting the specimen from the patient; A sample processing method using collecting a specimen using the specimen collection tool; introducing the specimen collected by the specimen collecting tool into the pre-test treatment container; a step of connecting the pre-inspection treatment container nozzle to the pre-inspection treatment container; Pressurizing the pre-inspection treatment container to drip the liquid contained in the pre-inspection treatment container using the filter; A sample processing method comprising:

Images