Kit and method for self-collection of earwax and measurement of biomarkers using the earwax in situ

Abstract

The present invention relates to a kit and a method for self-collecting earwax and measuring biomarkers in situ using the earwax. The kit includes a device for collecting a sample and further includes a kit for directly measuring biomarkers of the earwax present in the kit being collected.

Description

Technical Field

[0001] The present invention relates to a method and kit for obtaining an earwax sample for measuring the concentrations of various biomarkers, and in particular, to a process and kit for obtaining an earwax sample and measuring various biomarkers using a portable kit.

Background Art

[0002] As described in International Publication No. 2019 / 123392 of the same applicant, when using a device for self-collecting the inventor's earwax or earwax, this specimen is reliable in terms of reflecting the chronic systemic levels of substances that vary greatly in a short period, such as cortisol and glucose. Similarly, the document describes an efficient and safe device for extracting an earwax sample for analysis.

[0003] In addition to the device for extracting the above sample, it would be highly desirable to have a device that enables direct measurement of biomarker levels from a device that extracts the sample in situ rather than in vitro. The above prior art does not describe or suggest any device or method for measuring biomarker levels from earwax.

[0004] In view of the above, the applicant has developed a kit including the above device for extracting an earwax sample and, additionally, a portable device that can use a battery or current as a power source for directly measuring biomarkers from the earwax present in the device for extracting the earwax sample.

[0005] The above kit comprises a device for sample extraction similar to that described in International Publication No. 2019 / 123392 and a portable device for measuring biomarkers configured to receive a sponge of the device for sample extraction pre-impregnated with an earwax sample. The device for measuring the biomarker uses a cylindrical multi-lateral flow biosensor with a plurality of different aligned assays. However, other biosensors such as those called "ammeter" may also be used. The proposed device for measuring the biomarker has a camera for collecting test images, and the images are transmitted to the information processing means. As a result, the information processing means can interpret the images received from the test using an immunochromatography program.

[0006] The information processing means has an optical signal detector including a camera. The test results are displayed on a liquid crystal display (LCD). In this way, the user can practically and quickly obtain the concentrations of various biomarkers from a biological sample of earwax taken by the same user.

Summary of the Invention

Problems to be Solved by the Invention

[0007] Therefore, the main object of the present invention is to provide a kit and method for self-collecting earwax and measuring biomarkers in situ from earwax.

[0008] Another main object of the present invention is to provide a kit and method of the above-mentioned nature, including a device for obtaining a sample and additionally a device for directly measuring biomarkers from earwax present in the collection kit.

Means for Solving the Problems

[0009] Another object of the present invention is that a device for measuring biomarkers is designed to receive a sponge of a device for sample extraction having a cerumen sample, and uses a cylindrical multiplex lateral flow biosensor having a plurality of different tests adjacent to each other. The device for measuring the marker has a camera for collecting an image of the test, and the image is transmitted to information processing means for interpreting the image received from the test using an immunochromatography program. However, other methods may also be used.

Brief Description of the Drawings

[0010] These and other objects and advantages of the present invention's kit and method for self-collecting cerumen and measuring biomarkers in situ from the cerumen will be apparent to those skilled in the art from the following detailed description of the embodiments of the present invention made with reference to the accompanying drawings.

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DETAILED DESCRIPTION OF THE INVENTION

[0011] Here, a kit for self - collection of earwax and in - situ measurement of biomarkers from the earwax will be described with respect to its preferred embodiments. The kit includes a device for extracting a sample similar to that described in International Publication No. WO 2019 / 123392, the description of which is included below for reference, and a handle (1) having a first end (2) and a second end (3), wherein the second end (3) may include a threaded portion (4) and has coupling means in a preferred embodiment of the present invention, the handle (1) A detachable head (tip) comprising a base (5) and an elongated sponge holder (6) that can be extended longitudinally and directly depends on the upper part of the base, wherein the lower part of the base has a shell including an internal thread (7) in a pattern for receiving the thread (4) of the handle (1), and the sponge of the support (6) has a star-shaped cross section, the detachable head (tip). A longitudinally positioned central casing (not shown), and an elongated sponge (8) having a star-shaped cross section for receiving the sponge holder (6) of the base (5), wherein the handle (1) and the base (5) can include any coupling means suitable for coupling the handle, such as a pressure joint (9), the elongated sponge (8). The sponge (8) is preferably made of cellulose and may be adhered to the sponge holder (6) using a non-allergenic adhesive.

[0012] A device for measuring biomarkers from an earwax sample obtained by a device for extracting a sample. A cubic housing (10) having an upper wall (11), a lower wall (not shown), a front wall (12), a rear wall (13), a right side wall (14), and a left side wall (15), wherein the upper wall (11) has a central circular opening (16) for receiving a cylindrical biosensor holder (17), the cubic housing (10). Information processing means (not shown) attached to the lower wall of the housing (10). A pedestal (18) fixed to the middle part of the surface of the lower wall of the housing (10), wherein the pedestal (18) has a height lower than the walls of the housing (10), the pedestal (18). Four cameras (19a, 19b, 19c, 19d) attached in the upper third of the pedestal (18), each camera (19a, 19b, 19c, 19d) being directed towards the middle parts of the right side wall (14), the left side wall (15), the corresponding front wall (12) and the rear wall (13), and each camera (19a, 19b, 19c, 19d) being connected to the information processing means, the four cameras (19a, 19b, 19c, 19d). Focusing means for enabling better resolution of an image obtained by a camera, the focusing means comprising a circular lens (20) surrounding a pedestal (18) having a camera (19a, 19b, 19c, 19d) and a cylindrical biosensor holder (17), and being held by support means (not shown) fixed to the lower wall. A light source, and in a preferred embodiment, includes four light sources (21a, 21b, 21c, 21d) attached to the pedestal (18) below the camera (19a, 19b, 19c, 19d), each light source being connected to a power source (not shown), and each light source (21a, 21b, 21c, 21d) generating light having specific monochromatic spectral characteristics within a wavelength range of 600 nm to 1400 nm to ensure a high contrast between the background and a specific staining zone provided by the reaction in each of the four biosensors described later. At least one screen (P) for presenting information, each being connected to information processing means for presenting the measurement results. A biosensor holder (17) comprising a hollow cylindrical element, the hollow cylindrical element having a cylindrical wall with an outer surface (22) and an inner surface (not shown), an upper end (23) and a lower end (24), and a wall of an upper end (25) having a diameter larger than that of the cylindrical element, the lower end remaining open, the upper part of the cylindrical element having a slot (26) with a length of + / - 4 mm, and the cylindrical element having a diameter that enables it to closely pass through a central circular opening (16) of a device for measuring biomarkers. A cylindrical biosensor unit (27) having at least one multiplexed lateral flow biosensor (28) shown in FIG. 10, the biosensor unit (27) comprising the following. Other materials may also be used, such as vinyl, which has a reliable bond to proteins (including antibodies or enzymes), and a flexible backing plate (29) having a hollow cylindrical shape with an open end made of a plastic material such as nitrocellulose, which has variable absorption characteristics (different capillary flow times). The cylindrical backing plate has an outer surface, an inner surface, a diameter, and a height such that it can be tightly inserted into the biosensor holder (17), and the backing plate has stop means (30) on the upper portion of the outer surface that engage within the slot (26) of the biosensor holder (17) when the backing plate (29) is inserted into the biosensor holder (17). One or more multiplex lateral flow biosensors (28) each positioned equidistantly on the intermediate portion of the inner surface of the flexible backing plate (29) following the diameter of the backing plate. Each multiplex lateral flow biosensor comprises the following. An elongated base membrane (31) made of a flexible material selected from a set including vinyl or nitrocellulose. The base membrane is made of acrylic but is pressure sensitive by use of a non-reactive and stable adhesive. The base membrane (31) has a first end (32), a second end (33), an inner surface, and an outer surface (34), and the inner surface can have hydrophilic properties to slow down the diffusion of a soiled (hydrophobic) sample, thus ensuring the time required for an immunohistochemical reaction to occur. An elongated nitrocellulose membrane (35) having a first end (36) and a second end (37) and having a test line portion (38) and a control line portion (39). The nitrocellulose membrane (35) is disposed vertically along its entire length on the base membrane (31), and its first end (36) and second end (37) coincide with the first end (32) and second end (33) of the base membrane (31). A conjugate pad (40) disposed vertically at its first horizontal end (36) on the nitrocellulose membrane to contain a labeled tracer antibody.

[0013] The cerumen sample pad (41) is disposed vertically on the conjugate pad (40). The sample pad (41) is made of a highly oil-absorbent material such as natural inorganic substances such as sand, clay, or volcanic ash. It can also be made of other natural organic materials and synthetic materials and must be pretreated with a buffer solution. The absorption pad (42) is disposed vertically at its second horizontal end (37) on the nitrocellulose membrane (35).

[0014] The components of each biosensor (28) are attached to the inner surface of the backing plate (29) of the biosensor unit (27) using an adhesive substance. Next, each component of each biosensor (28) is attached to the base membrane (31) (also called the test strip) in close contact with each other so that the movement of the sample and test components through the biosensor (28) is ensured during the test. Preferably, it overlaps with the end that binds to another component by about 2 mm.

[0015] Cerumen (ear wax) diffuses into the sample pad (41), and the sample pad acts as a filter to facilitate the flow of the sample and test components. The conjugate (enzyme or antibody) used is dispensed onto the conjugate pad (40) with a pipette. In other embodiments, the pads may be prepared separately in advance, dried at room temperature, and stored in a desiccator at 4°C before use and attachment to the biosensor. The corresponding capture reagent is dispensed onto the membrane. Nitrocellulose (35). The test line (38) and the control line (39) are each arranged by any device or suitable method.

[0016] The label (tracer) is disposed on the pad and the nitrocellulose membrane and is preferably made of colloidal gold, carbon, or latex. A specific tag is selected based on its compatibility with the test components. When specific analytes in the cerumen reach the conjugate pad (40), they rehydrate the labeled tracer antibody to collate with each other (zone (A) shown in FIG. 10).

[0017] The resulting complex moves from the region marked (A) to the region marked (D) through the principle of capillary action until it is detected on the test line (38) via the pre-deposited detection antibody (zone (B)). If there is no conjugation on the test line (38), a negative result is indicated.

[0018] Figure 11a shows the potential results (staining) provided by each biosensor (28), which indicates a positive result, Figure 11b shows a negative result, and Figure 11c shows an invalid result. On the test line (38), the gold-labeled antibody recognizes and binds to a specific portion of the target molecule known as an epitope.

[0019] The unreacted antibody / tracer conjugate is ultimately detected at the control line (39) by the species-specific antibody (zone (C)). Both the test line (38) and the control line (39) contain a capture reagent for indicating the result. The visual control line (39) indicates that the test was performed correctly. The label colors the control line (39) and the test line (38). Finally, the presence of the absorption pad (42) collects any excess sample and reagent (zone (D)).

[0020] To conduct the test, a cylindrical biosensor unit (27) having one or more (preferably up to four) multiplex lateral flow biosensors (28) and having an earwax sample, which is prepared in advance, is inserted into a biosensor holder (17). The biosensor holder (17) is inserted into a central circular opening (16) of a device for measuring a biomarker such that the upper end wall (25) of the biosensor holder (17) functions as a stop against the upper wall (11) of the device for measuring the biomarker. When the biosensor holder (17) is inserted into the device for measuring the biomarker, the inner surface of the backing plate (29) of the biosensor unit (27) surrounds a pedestal (18) having cameras (19a, 19b, 19c, 19d) such that each chamber (19a, 19b, 19c, 19d) points to a test line (38) and a control line (39), also called a test strip or a staining zone, of the multiplex lateral flow biosensor (28).

[0021] Each camera (19a, 19b, 19c, 19d) obtains images of the test line (38) and the control line (39) (staining zone) as data from a matrix of points arranged in horizontal and vertical columns. Each point or pixel is characterized by three numerical parameters corresponding to three image-forming channels that make up an RGB circuit, namely red, green, and blue (RGB). The recorded result of each channel is a number that increases as the staining intensity increases.

[0022] The images obtained by each camera (19a, 19b, 19c, 19d) are processed in an information processing means by immunochromatography software that tests four readings corresponding to the chronic levels of the substances measured in the earwax. The results are shown on a screen (P) each corresponding to a camera (19a, 19b, 19c, 19d) and its corresponding biosensor (28). In other embodiments, the software that plays the role of processing information from the cameras (19a, 19b, 19c, 19d) may include an artificial intelligence algorithm that can be trained to improve the interpretation of the obtained images and thus the measurement of the biomarker.

[0023] Use of the kit Prior to this, the user must prepare a multiplex lateral flow biosensor (28) as follows. · Apply a conjugate (enzyme or antibody) to the conjugate pad (40). · Apply the corresponding capture reagent onto the nitrocellulose membrane (35) where the test line (38) and the control line (39) are respectively arranged.

[0024] Insert the biosensor unit (27) cylindrically into the biosensor holder (17) such that the stop means (30) of the backing plate (29) catches in the slot (26) of the biosensor holder, and fix the biosensor unit (27) inside the biosensor holder (17).

[0025] Sample extraction is preferably performed using a sample extraction device similar to that described in WO 2019 / 123392, the description of which is incorporated herein by reference. However, any other type of device can be used for extraction as long as it is compatible with the shape and size of the cylindrical opening of the biosensor unit (27) inserted into the biosensor holder (17) and has a sponge with a length sufficient to contact a sufficient amount of sample with the sample pad (41) of each biosensor (28). In a preferred embodiment, it is possible to analyze cortisol and glucose from all previously obtained cerumen samples with a small cerumen sample of 0.8 mg, and thus the minimum sample amount is preferably 0.8 mg.

[0026] Cerumen is obtained by inserting the sponge tip (8) of the collection device into the external auditory canal and rotating the sponge in the external auditory canal for about 30 - 60 seconds. Once a cerumen sample is obtained, the user must insert the sponge tip of the sample extraction device into the cylindrical biosensor unit located inside the biosensor holder as shown in FIG. 12.

[0027] Inside the biosensor unit (27), the user must rotate the sponge (8) so that the sponge (8) rubs against the sample pad (41) of each biosensor (28) and deposits a biological earwax sample (at least preferably 0.8 mg) in each sample pad (41).

[0028] When the sample pad (41) is impregnated with the earwax sample, the biosensor unit (27) must be inserted into the central circular opening (16) of the biomarker measurement device so that the upper end wall (25) of the biosensor holder serves as a stop with the upper wall (11) of the device for measuring the biomarker, as shown in FIG. 13.

[0029] To ensure that each visual control line (38), (39) is aligned with its respective camera (19a, 19b, 19c, 19d), the biosensor holder (17) may include guiding means (not shown) that cooperate with the circular central opening (16) of the housing.

[0030] As described above, the device for measuring a biomarker uses the conversion of different electrochemical signals as a reading mechanism for different biomarkers such as glucose present in human earwax. However, other optical reading mechanisms may also be used.

[0031] The degree of fluorescence determines the specific electrical signal and concentration of the analyte to be determined, such as glucose, via a specific calibration curve stored in the memory of the device. Other conjugates such as gold or silver nanoparticles that emit different wavelengths that can also be converted to different concentrations may also be used.

[0032] In other embodiments, the device for measuring a biomarker of the present invention can use different types of biosensors such as, but not limited to, amperometric, potentiometric, impedance, voltammetric, piezoelectric, thermometric, optical, etc. biosensors.

[0033] Some of the above biosensors can provide measurement results through signals transmitted to the information processing unit either directly or wirelessly, so a camera is not required. Therefore, in the above modality, a camera is not needed. There is no need to use a pedestal together with a camera, lighting, or focusing means. Similarly, it is possible that the biosensor used does not require prior preparation by the user (administering the conjugate to be used and administering the corresponding capture reagent).

[0034] Similarly, in other embodiments, the cross-section of the sponge (8) of the device for sample extraction, the central opening (16) of the device for measuring biomarkers, and the biosensor holder (17) and biosensor unit (27) can have any cross-sectional shape as long as the sponge (8) can deposit an appropriate amount of earwax on the biosensor. Furthermore, the biosensor holder (17) can comprise only the structure and means for holding one or more sensors.

[0035] Similarly, the housing (10) of the device for measuring biomarkers can have any cross-sectional shape such as circular, or can even include only the structure having means for holding the biosensor holder (17).

[0036] Finally, in other embodiments, instead of using the biosensor holder (17), only the biosensor unit (27) may be inserted into the device for measuring biomarkers. The unit must have a cavity so that the sponge of the sampling device can be inserted to impregnate the sensor with the earwax sample.

[0037] Finally, the kits and methods for self-collection of earwax and in situ biomarker measurement from the earwax of the present invention are not limited to the modalities described above, and those skilled in the art are qualified to make changes to the kits and methods for self-collection of earwax and in situ biomarker measurement from the earwax of the present invention in accordance with the teachings established herein, the scope of which is exclusively established by the appended claims and should be understood accordingly.

Claims

1. A device for measuring biomarkers from an earwax sample obtained by a sample extraction device, the sample extraction device comprising a handle and a head attached to the handle, the head having a sponge holder and a sponge attached to the sponge holder, the sponge being for collecting earwax by insertion into the ear canal, the device for measuring the biomarker comprising: A casing having at least one upper wall, the upper wall having a central opening; Information processing means mounted within an inner portion of the housing; At least one screen for presenting information, each connected to the information processing means for presenting the results of biomarker measurements; One or more biosensors each disposed within the housing, the sponge of the sample extraction device being contactable such that the sponge can deposit a sample of earwax within each biosensor; A device for measuring biomarkers, comprising.

2. A lower wall; A camera holder pedestal fixed to an intermediate portion of the surface of the lower wall of the housing; At least one camera attached to an upper portion of the pedestal, each camera being arranged to face a respective biosensor, each camera being connected to the information processing means; Focusing means surrounding the pedestal and the one or more cameras to enable better resolution of the images obtained by the cameras; A light source disposed within the housing; The device for measuring biomarkers according to claim 1, further comprising.

3. The focusing means includes a circular lens that surrounds a pedestal having the camera and the cylindrical biosensor and is held by a support means fixed to the lower wall, and the light source generates optical illumination having a monochromatic spectral characteristic within a wavelength range of 600 nm to 1400 nm. A device for measuring a biomarker according to claim 2.

4. One or more biosensors are attached to a removable biosensor unit, and the biosensor unit A flexible backing plate having a hollow cylindrical shape with an open end made of a material plastic having strong binding to proteins containing antibodies or enzymes, such as nitrocellulose, and variable absorption characteristics (different capillary flow times), the cylindrical backing plate having an outer surface and an inner surface, and each biosensor being attached to the inner surface of the backing plate. The biosensor unit is inserted into a device for measuring the biomarker through the central opening. A device for measuring a biomarker according to claim 1.

5. The central opening of the housing has a cylindrical shape, and the device for measuring the biomarker Further includes a biosensor holder having a hollow cylindrical element, the hollow cylindrical element having a cylindrical wall having an outer surface and an inner surface, an upper end and a lower end, and an upper end wall having a diameter larger than that of the cylindrical element, the lower end remaining open, and the cylindrical element having a diameter that allows it to fit tightly into the circular central opening of the device for measuring the biomarker. One or more biosensors are attached to a biosensor unit, and the biosensor unit comprises a hollow cylindrical flexible backing plate with an open end made of a plastic material having a strong binding to proteins including antibodies or enzymes such as nitrocellulose. The cylindrical backing plate has an outer surface, an inner surface, a diameter, and a height such that it can be tightly inserted into a biosensor holder. Each biosensor is attached to the inner surface of the backing plate. The biosensor unit is inserted into the biosensor holder, and the biosensor holder is inserted into the central opening of the housing having a cylindrical shape. A device for measuring a biomarker according to claim 1.

6. The central opening of the housing has a cylindrical shape, and the device for measuring the biomarker a lower wall, a camera holder pedestal fixed to an intermediate portion of the bottom wall surface of the casing, at least one camera attached to an upper portion of the pedestal, each camera being arranged to face a respective biosensor, and each camera being connected to the information processing means. Focusing means surrounding the pedestal and one or more cameras to enable better resolution of the images obtained by the cameras. a light source disposed inside the housing, a biosensor holder comprising a hollow cylindrical element, the hollow cylindrical element having a cylindrical wall with an outer surface and an inner surface, an upper end and a lower end, and an upper end wall having a diameter larger than that of the cylindrical element, the lower end being open, and the cylindrical element having a diameter such that it can tightly pass through the circular central opening of the device for measuring the biomarker. One or more biosensors are attached to a biosensor unit, and the biosensor unit comprises a hollow cylindrical flexible backing plate with an open end made of a plastic material having a strong binding to proteins including antibodies or enzymes such as nitrocellulose, the cylindrical backing plate having an outer surface, an inner surface, a diameter and a height such that it can be tightly inserted into the biosensor holder, each biosensor being attached to the inner surface of the backing plate, the biosensor unit being inserted into the biosensor holder, the biosensor holder being inserted into the central opening of the casing having a cylindrical shape, Each biosensor has a reactive strip (staining zone), The upper end wall of the biosensor holder functions as a contact portion with the upper wall of the device for measuring the biomarker, When the biosensor holder is inserted into the device for measuring the biomarker, the inner surface of the backing plate of the biosensor unit surrounds the pedestal having the camera such that each camera points to the test strip (staining area) of the multiplex lateral flow biosensor, The images obtained by each camera are processed in an information processing medium through immunochromatography software that tests four readings corresponding to the chronic levels of the substances measured in the earwax, and the results are shown on the screen, each corresponding to a camera and its corresponding biosensor, of the device for measuring the biomarker according to claim 1.

7. Each sensor is a multiplex lateral flow biosensor, and the multiplex lateral flow biosensor is, Although not limiting, an elongated base film made of a flexible material selected from a set including vinyl or nitrocellulose, wherein the base film is acrylic but pressure-sensitive by the use of a non-reactive and stable adhesive, the base film having a first end, a second end, an inner surface, and an outer surface, the inner surface capable of having hydrophilic properties to slow the diffusion of earwax samples (hydrophobic), thus ensuring the time required for an immunohistochemical reaction to occur, a base film, and An elongated nitrocellulose membrane having a first end and a second end and having a test line portion and a control line portion, the nitrocellulose membrane being disposed vertically on the base film along its entire length, the first end and the second end thereof coinciding with the first end and the second end of the base film, a nitrocellulose membrane, and A conjugate pad disposed vertically at its first horizontal end on the nitrocellulose membrane for containing a labeled tracer antibody, and An earwax sample pad disposed vertically on the conjugate pad, the sample pad being made of a highly oil-absorbent material such as natural inorganic substances such as sand, clay, or volcanic ash, which can also be made of other natural organic materials and synthetic materials and must be pretreated with a buffer solution, an earwax sample pad, and An absorption pad disposed vertically at its second horizontal end on the nitrocellulose membrane, and wherein the earwax diffuses into the sample pad and the sample pad also acts as a filter, and The conjugate (enzyme or antibody) used is dispensed onto the conjugate pad, and The corresponding capture reagents are respectively dispensed onto the nitrocellulose membrane where the test line and the control line are located, and Labels (tracers) disposed on the pads and the nitrocellulose membrane, made of materials selected from but not limited to colloidal gold, carbon, or latex, and The device for measuring a biomarker according to claim 6, wherein the complex obtained in the analysis moves by the principle of capillary action until it is detected on the test line via a pre-deposited detection antibody.

8. A kit for self-collecting earwax and measuring a biomarker in situ from the earwax, A device for sample extraction, comprising a handle and a head attached to the handle, the head having a sponge holder and a sponge attached to the sponge holder for collecting earwax by insertion into the external auditory canal, a device for sample extraction, A device for measuring a biomarker from an earwax sample obtained by the sample extraction device according to any one of claims 1 to 7 and A kit comprising the same.

9. A method for self-collecting earwax and measuring a biomarker in situ from the earwax, a. Extracting a sample of earwax using a device for removing earwax, comprising a handle and a head attached to the handle, the head having a sponge holder and a sponge attached to the sponge holder for obtaining earwax by insertion into the external auditory canal; b. Depositing the earwax using the sponge of the device for removing earwax in one or more biosensors of the device for measuring a biomarker according to any one of claims 1 to 7; A method comprising the same.

10. A multiplex lateral flow biosensor, An elongated base film made of a flexible material selected from a set including vinyl or nitrocellulose, although not limiting, wherein the base film is acrylic but pressure-sensitive by a non-reactive and stable adhesive, the base film having a first end, a second end, an inner surface, and an outer surface, the inner surface being capable of having hydrophilic properties for slowing the diffusion of an earwax sample (hydrophobic), thus ensuring the time required for an immunohistochemical reaction to occur, a base film, and An elongated nitrocellulose membrane having a first end and a second end and having a test line portion and a control line portion, the nitrocellulose membrane being disposed vertically along its entire length on the base film, the first end and the second end thereof coinciding with the first end and the second end of the base film, a nitrocellulose membrane, and A conjugate pad disposed vertically at its first horizontal end on the nitrocellulose membrane for containing a labeled tracer antibody, and An earwax sample pad disposed vertically on the conjugate pad, the sample pad being made of a highly oil-absorbing material such as natural inorganic substances such as sand, clay, or volcanic ash, which can also be made of other natural organic materials and synthetic materials and must be pretreated with a buffer solution, an earwax sample pad, and An absorption pad disposed vertically at its second horizontal end on the nitrocellulose membrane, and wherein the earwax diffuses into the sample pad and the sample pad also acts as a filter, and The conjugate (enzyme or antibody) used is dispensed onto the conjugate pad, and The corresponding capture reagents are respectively dispensed onto the nitrocellulose membrane where the test line and the control line are located, and Including labels (tracers) disposed on the pads and the nitrocellulose membrane, made of materials selected from but not limited to colloidal gold, carbon, or latex, and A multiplex lateral flow biosensor in which the complex obtained in the analysis moves by the principle of capillary action until it is detected on the test line through a pre-deposited detection antibody.

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