Detection device, mask used in detection device, identification means used in detection device, visualization means used in detection device, detection method, detection kit, and detection system

Abstract

Based on the inventor's previous knowledge, this project aims to establish a novel detection and inspection method that utilizes nonwoven fabric, and in particular, a detection and inspection method using a wearable nonwoven fabric mask. [Solution] Pathogens are detected by capturing them from an infected person using a nonwoven fabric, specifically a mask.

Description

Technical Field

[0001] The present invention relates to a detection device, a mask used in the detection device, an identification means used in the detection device, a visualization means used in the detection device, a detection method, a detection kit, and a detection system, and particularly to a detection device that captures a pathogen from an infected person infected with the pathogen on a non-woven fabric and detects the pathogen, etc.

Background Art

[0002] As means for inspecting viruses, various inspection kits have been proposed in addition to large-scale inspection means such as PCR, but there is no means for detecting and inspecting viruses after carrying an antibody on a non-woven fabric.

Summary of the Invention

Problems to be Solved by the Invention

[0003] By the way, the inventor of the present application has deep knowledge of the technology using the ostrich antibody, developed a filter carrying the ostrich antibody on a non-woven fabric, and also commercialized an ostrich antibody mask from the development results.

[0004] Therefore, an object of the present invention is to establish a new detection and inspection means that has not existed until now in terms of using a non-woven fabric based on the knowledge of the inventors so far, and particularly to establish a detection and inspection means using a wearable such as a non-woven fabric mask.

Means for Solving the Problems

[0005] A first aspect of the present invention is a detection device that captures a pathogen from an infected person infected with the pathogen on a non-woven fabric and detects the pathogen.

[0006] In a second aspect of the present invention, in the first aspect, the pathogen is SARS-CoV-2 (novel coronavirus).

[0007] In a third aspect of the present invention, in the first or second aspect, the nonwoven fabric is a mask.

[0008] In a fourth aspect of the present invention, in the third aspect, the mask has a filter on which an antibody is supported.

[0009] In a fifth aspect of the present invention, in the fourth aspect, the antibody is an animal-derived antibody.

[0010] In a sixth aspect of the present invention, in the fifth aspect, the animal-derived antibody is an ostrich antibody.

[0011] A seventh aspect of the present invention is a detection device for detecting a pathogen by capturing the pathogen on a nonwoven fabric from an infected person, wherein the nonwoven fabric is a mask, the mask includes a filter carrying antibodies, and the mask is equipped with identification means for the infected person to wear the mask and for the antibodies on the filter to identify the pathogen captured from the infected person's mouth or nose.

[0012] In the eighth aspect of the present invention, in the seventh aspect, the identification means is a secondary antibody labeled with a fluorescent or luminescent dye or enzyme.

[0013] A ninth aspect of the present invention is that, in the seventh or eighth aspect, the invention is further comprising visualization means for visualizing and detecting the pathogen identified by the identification means.

[0014] In the tenth aspect of the present invention, in the ninth aspect, the visualization means is a light source that emits light.

[0015] In the eleventh aspect of the present invention, in the tenth aspect, the light source is capable of irradiating light with wavelengths of 260 nm to 600 nm.

[0016] In a twelfth aspect of the present invention, in the eleventh aspect, the light source is the LED light of a smartphone.

[0017] A thirteenth aspect of the present invention is a mask used in a detection device described in any of the third to twelfth aspects.

[0018] A fourteenth aspect of the present invention is a mask used in a detection device described in any of the fourth to twelfth aspects, wherein the antibody-supported filter consists of one or more layers and can be peeled off from the surface.

[0019] A fifteenth aspect of the present invention is an identification means used in the detection device described in the seventh or eighth aspect.

[0020] A sixteenth aspect of the present invention is a visualization means used in a detection device described in any of the ninth to twelfth aspects.

[0021] A seventeenth aspect of the present invention is a detection method for detecting a pathogen by capturing it on a nonwoven fabric from an infected person.

[0022] The eighteenth aspect of the present invention is a detection kit for detecting a pathogen from an infected person, comprising a filter carrying ostrich antibodies, a nonwoven mask worn by the infected person, and identification means for identifying the pathogen captured by the ostrich antibodies in a way that makes it visible.

[0023] A 19th aspect of the present invention is a detection system for detecting a pathogen from an infected person, comprising: a nonwoven mask worn by the infected person, which includes a filter carrying ostrich antibodies; an identification means for identifying the pathogen captured by the ostrich antibodies so that it can be visualized; a light source for irradiating light to visualize and detect the pathogen identified by the identification means; and a database for accumulating data by linking the detection result of whether or not the pathogen was detected by the light source with the infected person.

[0024] The 20th aspect of the present invention, in the 19th aspect, comprises analysis means for analyzing the data stored in the database.

Advantages of the Invention

[0025] According to the invention of the present application, based on the findings of inventors to date, it is possible to establish a new detection and inspection means that has not existed until now in terms of using non-woven fabrics, and in particular, it is possible to establish a detection and inspection means using a wearable such as a non-woven mask.

Brief Description of the Drawings

[0026] [Figure 1] It is a diagram for explaining a detection method which is an example of an embodiment of the invention of the present application. [Figure 2] It is a diagram for explaining the result obtained by the detection method which is an example of an embodiment of the invention of the present application.

Modes for Carrying Out the Invention

[0027] Hereinafter, embodiments of the invention of the present application will be described with reference to the drawings. Note that the invention of the present application is not limited to this embodiment.

[0028] FIG. 1 is a diagram for explaining a detection method which is an example of an embodiment of the invention of the present application. FIG. 1(A) shows the state of removing the mask. Note that the crane antibody for corona is already bound to the mouth filter before using the mask. FIG. 1(B) shows the state of disinfecting the mask. The disinfection here is positioned as a process necessary for detoxifying the virus. FIG. 1(C) shows the state of peeling the mouth filter. FIG. 1(D) shows the state of taking out the mouth filter. FIG. 1(E) shows the state of spraying using a spray of a crane antibody (labeled secondary antibody for novel coronavirus) fluorescently labeled with FITC, rhodamine, etc. FIG. 1(F) shows the state of installing the mouth filter on the light irradiation device. [Outline of Research]

[0029] 1. Production of spike proteins (antigens) of coronavirus variant strains The gene for the spike protein of a variant of the novel coronavirus was incorporated into an E. coli vector and purified in large quantities. This recombinant protein was then introduced into human HEK cells to purify it. This recombinant protein was then immunized (injected) into ostriches, and antibodies were recovered from the ostrich egg yolks. The reactivity, specificity, and performance of the obtained antibodies were measured using ELISA and viral infection experiments to confirm their usefulness.

[0030] 2. Creation of an ostrich antibody-supported filter that maximizes the trapping of the novel coronavirus. We developed filters that can maximize the activity of mass-produced ostrich antibodies using methods such as physically loading antibodies onto nonwoven fabric and covalently binding antibodies with polylactic acid. Since a liquid phase is necessary for virus visualization, we optimized the filters to be able to trap even the smallest amount of virus by varying the amount of antibody loaded on the filter material while verifying the antibody retention of the filter material using ELISA.

[0031] 3. Preparation of secondary antibodies to be used as probes To visualize the virus particles captured by the ostrich antibody-supported filter, we prepared secondary antibodies (ostrich polyclonal antibodies that recognize coronavirus) labeled with fluorescent / luminescent dyes or enzymes, and selected labeling methods and substrates that allow for visual determination of color development and fluorescence.

[0032] 4. Visualization of COVID-19 We successfully visualized the novel coronavirus by reacting a secondary antibody with an ostrich antibody-loaded filter containing liquefied viral antigen in the laboratory, and with an ostrich antibody-loaded mask (mouth filter) used by a person infected with the novel coronavirus, and then irradiating it with light of a specific wavelength. We also confirmed that the virus on the ostrich antibody-loaded filter could be visualized even when using the LED light from a SmartFin as one of the light sources.

[0033] Figure 2 is a diagram illustrating the results obtained by a detection method, which is an example of an embodiment of the proposed invention. Figures 2(A) to 2(C) show experiments with a filter pre-dropped with the spike protein of the novel coronavirus, and Figure 2(D) shows the results with a mask used by a person infected with the novel coronavirus for 8 hours. Figure 2(A) shows the state of the mask (mouth filter) before light irradiation (see Figure 1(F)). Here, the location of the coronavirus is unknown. Figure 2(B) shows the state of the mask (mouth filter) during light irradiation, specifically the case of spraying with FITC-labeled ostrich antibody, where six areas (green fluorescent areas) are the locations of the coronavirus antigen. Excitation light with a peak wavelength of 350 nm was used for visualization. Figure 2(C) shows the state of the mask (mouth filter) during light irradiation, specifically the case of spraying with rhodamine-labeled ostrich antibody, where the areas that stand out as a pattern (red fluorescent areas) are the locations of the coronavirus antigen. Excitation light with a peak wavelength of 540 nm was used for visualization. Figure 2(D) shows the state of the mask (mouth filter) during light irradiation, specifically the case of spraying FITC-labeled ostrich antibody. The dashed lines correspond to the position of the nose, and the image shows that a large amount of virus is attached to both sides of the nose (green fluorescent areas). Excitation light with a peak wavelength of 350 nm was used for visualization. For the visualization of pathogens, light with a peak wavelength between 260 nm and 600 nm is preferable, for example.

[0034] Figure 2 shows that ostrich antibodies for coronavirus placed on the mouth filter of the mask capture coronavirus in coughs, sneezes, and nasal discharge. In addition, it is shown that the virus can be visualized by reacting fluorescently labeled ostrich antibodies (secondary antibodies) and irradiating them with light. From these results, the inventors have found that by using an ostrich antibody-carrying mask, a nonwoven fabric mask with a mouth filter carrying ostrich antibodies, it is possible to visualize SARS-CoV-2 (novel coronavirus) from exhaled breath using the fluorescent antibody method, even with the naked eye.

[0035] In particular, as shown in Figure 2(D), the presence of the virus was confirmed even in an ostrich antibody-loaded mask (clinical sample) used for 8 hours by a person infected with the novel coronavirus. Furthermore, as shown in Figure 2, we succeeded in developing a technique to visualize (visually observe) the virus captured in masks used by infected individuals using a light source box.

[0036] Furthermore, when using the LED light from a smartphone as the light source, we also succeeded in visualizing the viral antigen on the ostrich antibody-supported filter.

[0037] Going forward, as the inventor, I plan to accelerate research on developing nonwoven fabric filters with extremely low autofluorescence (background noise), selecting light wavelengths to strongly visualize viral antigens captured by ostrich antibody-supported masks, establishing a method for detecting viruses including mutant strains using ostrich antibody-supported masks and smartphone LED light, and working towards the practical application of a technology that can rapidly and inexpensively detect viral shedding from asymptomatic infected individuals.

[0038] The significance of the invention is summarized below. In COVID-19, the number of people infected by one person (effective reproduction number Rt) is not very large. However, there are superspreaders who release large amounts of the virus, about 1 in 10 infected individuals, and a single person can infect many people. Therefore, detecting the virus in the breath of asymptomatic carriers is an effective means of preventing outbreaks.

[0039] The inventor combined his world-first "low-cost mass production technology for highly sensitive COVID-19 antibodies using ostriches" with "antibody loading technology onto fiber materials" to develop a virus detection technology that requires only simple light irradiation. This technology can simultaneously bind antibodies to not only coronaviruses but also influenza and mycoplasmas, and by changing the fluorescent dye used to label the secondary antibodies, it is possible to distinguish between various pathogens by color differences in a single test.

[0040] If viral infections can be detected using disposable masks, which are used daily all over the world, asymptomatic carriers (such as superspreaders) can be isolated early on, which in turn can prevent outbreaks and household transmissions.

[0041] In the future, the inventor plans to develop a mask for simple and rapid measurement of viruses in exhaled breath, create a wearable testing kit using LED light from a smartphone, and begin creating a database of biological responses to COVID-19 infection during smartphone facial recognition. Furthermore, based on the analysis of this data, they also plan to commercialize AI-powered infectious disease diagnostic technology using wearable devices (smartphones, watches, glasses, rings, etc.).

[0042] The inventor believes that the impact of this research on society will be that, through the research described above, it will lead to the early social implementation of a new technology for mass disease control in a post-COVID-19 society, specifically by preventing asymptomatic carriers (such as superspreaders), who are a major factor in the spread of COVID-19, from entering groups.

[0043] In the above embodiment, a mask was given as an example of nonwoven fabric, but other nonwoven fabrics such as wet wipes may also be used.

[0044] Furthermore, in the above embodiment, ostrich antibodies were given as an example, but antibodies derived from other animals may also be used, or any other antibody regardless of whether it is of animal origin.

[0045] Furthermore, in the above embodiment, a mask was used in which the mouth filter could be peeled back to facilitate peeling, but it is not necessary for the filter to be peeled back or not. If it is peeled back, for example, if the antibody-supported filter consists of multiple layers and can be peeled back sequentially from the surface, it becomes possible to perform multiple detection and testing tests, which improves the convenience of use for the user.

[0046] By the way, regarding how to provide the product to users, the mask and identification spray can be sold as a set kit, or the identification spray can be sold separately. If a smartphone is used as the light source, the above sales are likely sufficient, but the light source can also be sold separately. If the light source is sold separately, it can be sold as a set kit, or sold separately.

[0047] Furthermore, by linking smartphone personal authentication with virus detection results from masks, it may be possible to accumulate data on infected individuals. However, the data from smartphones could also be handled by a system using an app that links the smartphone's LED light and camera to capture images, which are then sent via email along with personal information obtained through personal authentication. If data is only sent when detection occurs, an app to analyze the detection results would be needed on the smartphone side. However, if detection is performed on the server side, this is not necessarily required, and all captured photos can be sent. In any case, the server side will collect big data, which can then be analyzed using AI and other technologies, enabling various types of analysis.

Claims

1. A detection kit that visualizes and detects pathogens, A mask having a nonwoven fabric filter on which ostrich antibodies are carried to capture the pathogen from the mouth or nose of an infected person infected with the pathogen, The system includes an identification means worn by the infected person, which allows the ostrich antibodies in the filter to identify the pathogen captured from the infected person's mouth or nose. The identification means is a detection kit comprising an antibody that recognizes the pathogen, labeled with a fluorescent or luminescent dye or enzyme.

2. The detection kit according to claim 1 or 2, wherein the ostrich antibody that recognizes the pathogen is an ostrich polyclonal antibody.

3. A mask used in the detection kit according to claim 1 or 2.

4. The mask according to claim 3, wherein the nonwoven fabric filter on which the ostrich antibody is supported consists of a single phase or multiple layers and can be peeled off from the surface side.

5. An identification means used in the detection kit according to claim 1 or 2.

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