Cerumen self-collection and in-situ biomarker measurement suite and method

Abstract

Each cerumen self-collection and in-situ biomarker measurement kit and method comprises a device for collecting a cerumen sample and is provided with a kit for directly measuring a biomarker from the cerumen sample of the collection device.

Description

Background of the Invention

[0002] A. Field of the Invention

[0003] The present invention relates to a method and kit for collecting cerumen samples to measure the concentrations of various biomarkers. This portable kit and method can be carried around for cerumen sample collection and measurement of multiple biomarkers.

[0004] B. Description of the Prior Art

[0005] As described in the document with the international patent application number WO2019 / 123392A1 by the same applicant, cerumen, i.e., earwax, is a reliable sample that can reflect the chronic systemic levels of certain substances, such as substances with large short-term variations like cortisol and glucose. At the same time, this document also describes an efficient and safe device for extracting cerumen samples for analysis.

[0006] In addition to the above-mentioned device for sample extraction, it is highly necessary to develop a device that can directly measure the biomarker levels in situ from the extraction device, thus avoiding measurement in test tubes. However, no device or method for measuring biomarker levels from cerumen has been described or proposed in the prior art.

[0007] In view of the above situation, the applicant has developed a kit, including a device for extracting cerumen samples and a portable device that uses a battery or power supply as an energy source to directly measure the sample biomarkers from the cerumen extraction device.

[0008] This kit mainly consists of a sample extraction device similar to that described in WO2019 / 123392A1 and a portable biomarker measurement device. This portable measurement device can process the sponge in the extraction device that has been pre-infiltrated with the cerumen sample. The biomarker measurement device uses a cylindrical multi-lateral flow biosensor, with multiple test sites arranged side by side on the sensor. In addition, other types of biosensors, such as "amperometric" biosensors, can also be used. The biomarker measurement device captures images of the test sites through a camera and transmits them to an information processing unit, which analyzes the obtained images through an immunochromatography program.

[0009] The information processing unit includes an optical signal detector composed of a camera device. The test results are displayed on an LCD liquid crystal display screen.

[0010] In this way, users can quickly and conveniently obtain the concentration information of various biomarkers through self-collected cerumen samples. Summary of the Invention

[0012] Therefore, the main object of the present invention is to provide a kit and method for self-collecting cerumen samples and in-situ biomarker measurement of cerumen.

[0013] Another main object of the present invention is to provide a kit and a method with the above functions, including a sample collection device and a kit for directly measuring biomarkers from the cerumen sample of the collection device.

[0014] This another main object of the present invention, that is, to provide a kit and a method with the above functions. Among them, the biomarker measurement device is designed to be able to receive a collection sponge with a cerumen sample and measure the sample through a cylindrical multiplex lateral flow biosensor with multiple side-by-side test sites. At the same time, the biomarker measurement device is equipped with a camera for capturing images of the test sites, and these images are transmitted to the information processing unit and interpreted and analyzed via an immunochromatography program. At the same time, other measurement methods can also be adopted.

[0015] The above and other objects and advantages of the present invention's self-collection of cerumen and in-situ biomarker measurement kit and method will be described in detail below in conjunction with the accompanying drawings, and those with ordinary skills in the relevant field can easily understand.

[0016] Brief description of the drawings

[0017] Figure 1 : Left side view of the cerumen extraction device of the present invention, including a cross-section of the tip with a sponge.

[0018] Figure 2 : Perspective view of the tip of the cerumen extraction device of the present invention, without the sponge.

[0019] Figure 3 : Top view of the tip of the cerumen extraction device of the present invention, without the sponge.

[0020] Figure 4 : Another left side view of the cerumen extraction device of the present invention, including a cross-section of the tip with a sponge.

[0021] Figure 5 : Perspective view of the biomarker measurement device of the present invention.

[0022] Figure 6 : Top view of the biomarker measurement device of the present invention, without the top housing.

[0023] Figure 7 : Front view of the biomarker measurement device of the present invention and its internal components, without the front housing.

[0024] Figure 8 : Front view of the biosensor bracket of the present invention.

[0025] Figure 9a : Perspective view of the biosensor unit of the present invention.

[0026] Figure 9b: Top view of the biosensor unit of the present invention.

[0027] Figure 10a : Front schematic view of the multiplex lateral flow biosensor assembly of the present invention.

[0028] Figure 10b : Perspective view of the biosensor unit of the present invention, excluding the outer front part, to show the multiplex lateral flow biosensor adhered to the inner side of the flexible outer plate.

[0029] Figure 11a 、 11b 、11c: Front view of the nitrocellulose membrane of the multiplex lateral biosensor of the present invention, showing the test line and control line in different states.

[0030] Figure 12 : Front view of the cerumen extraction device of the present invention, showing its sponge inserted into the biosensor unit, which is in turn inserted into the biosensor bracket.

[0031] Figure 13 : Perspective view of the biomarker measurement device of the present invention, including the biosensor bracket. The figure shows the biosensor unit about to be inserted into the measurement device through the central circular opening.

[0032] Detailed introduction of the invention

[0033] A kit for self - collection of cerumen and in - situ biomarker measurement will be described below and illustrated with reference to its preferred embodiments. The kit includes the following components:

[0034] The sample extraction device is similar to the device described in patent WO2019 / 123392A1, which is described as follows:

[0035] Handle (1): Comprises a first end (2) and a second end (3), where the second end (3) can be connected in different ways, and in the preferred embodiment, it can be a threaded connection (4).

[0036] Detachable head (tip): Includes a base (5) and a longitudinally extending sponge support (6) that is directly connected to the top of the base. There is a housing at the bottom of the base, and an internal threaded structure (7) for threaded connection (4) with the handle (1) is inside the housing. The cross - section of the sponge support (6) is star - shaped.

[0037] Long sponge (8): The sponge has a longitudinal housing (not shown in the figure) at its center, and its cross - section is star - shaped for accommodating the sponge support (6) on the base (5).

[0038] The handle (1) and the base (5) can be connected using any suitable connection method, such as a snap - fit connection (9).

[0039] The sponge (8) is preferably a cellulose sponge and is adhered to the sponge support (6) by an anti-allergy glue.

[0040] A device for measuring biomarkers of a cerumen sample obtained by a sample extraction device, comprising:

[0041] A cubic housing (10): including a top housing (11), a bottom housing (not shown in the figure), a front housing (12), a rear housing (13), a right housing (14), and a left housing (15). The top housing (11) is provided with a central circular opening (16) for inserting a cylindrical biosensor carrier (17).

[0042] An information processing unit (not shown in the figure): installed at the bottom of the cubic housing (10).

[0043] A pillar (18): fixed in the middle of the bottom surface of the cubic housing (10), and the height of the pillar (18) is lower than the height of the cubic housing (10).

[0044] Four cameras (19a, 19b, 19c, 19d): installed at one-third of the height above the pillar, and each camera (19a, 19b, 19c, 19d) faces the central area of the right housing (14), the left housing (15), the front housing (12), and the rear housing (13) respectively, and is connected to the information processing unit.

[0045] A focusing device: used to improve the resolution of the images captured by the cameras, including a circular lens (20) surrounding the cameras (19a, 19b, 19c, 19d), the pillar (18), and the cylindrical biosensor carrier (17), and is fixed by a support structure (not shown) fixed to the bottom housing.

[0046] A light source: In the preferred mode, it includes four light sources (21a, 21b, 21c, 21d) installed on the pillar (18), located below the cameras (19a, 19b, 19c, 19d). Each light source is connected to a power supply (not shown). Each light source (21a, 21b, 21c, 21d) generates monochromatic light with specific spectral characteristics, and the wavelength range is between 600nm and 1400nm to ensure a high contrast between the specific stained areas generated by the reactions of the biosensors and the background, which will be described in detail later.

[0047] At least one display screen (P): used to display information, and each display screen is connected to the information processing unit to present the measurement results.

[0048] A biosensor carrier (17), which is a hollow cylindrical structure. Its cylindrical outer shell includes an outer surface (22) and an inner surface (not shown), an upper end (23) and a lower end (24). The diameter of the outer shell (25) at the upper end is larger than that of the cylindrical body, and the lower end is open. There is a groove (26) with a length of approximately ±4 mm at the top of the cylindrical body. The diameter of the cylinder can just tightly pass through the central circular opening (16) of the biomarker measuring device.

[0049] A biosensor unit (27), which is a cylindrical structure and at least includes a multiplex lateral flow biosensor (28), as shown in Figure 10. The biosensor unit (27) includes:

[0050] A hollow cylindrical flexible outer plate (29), with both ends open, made of flexible materials such as vinyl polymers, or materials with strong binding force to proteins (including antibodies or enzymes) and different absorption characteristics (different capillary flow times), such as nitrocellulose. The outer surface and inner surface of the cylindrical outer plate have appropriate diameters and heights, and can just tightly be inserted into the biosensor carrier (17). There is a limiting device (30) above the outer surface of the top of the outer plate. When the outer plate (29) is inserted into the biosensor carrier (17), the limiting device (30) can be stuck into the groove (26) of the carrier to ensure that the outer plate (29) is fixed in the carrier (17).

[0051] One or more multiplex lateral flow biosensors (28): Each sensor is equally spaced in the middle part of the inner surface of the flexible outer plate (29) and arranged along the diameter of the outer plate. Each biosensor includes the following components:

[0052] An elongated base film (31), made of flexible materials, and the materials may include but are not limited to vinyl polymers or nitrocellulose. The base film is pressure-sensitive and is fixed by a stable acrylic adhesive without chemical reaction with the adhesive. The base film (31) includes one end (32), the other end (33), an inner surface and an outer surface (34). The inner surface has hydrophilic properties, which can slow down the diffusion of hydrophobic cerumen samples, thus ensuring the time required for immunohistochemical reactions.

[0053] An elongated nitrocellulose membrane (35), including one end (36), the other end (37), a test line area (38) and a control line area (39). The nitrocellulose membrane (35) is vertically fixed above the base film (31) and extends along its entire length. One end (36) and the other end (37) of it are flush with one end (32) and the other end (33) of the base film (31) respectively.

[0054] A conjugate pad (40): Vertically fixed in the horizontal direction at one end (36) of the nitrocellulose membrane for accommodating conjugate tracer antibodies.

[0055] Cerumen sample pad (41): Vertically fixed above the conjugate pad (40) for receiving cerumen samples. The material of the sample pad (41) has a high absorption capacity for grease and can be natural inorganic materials such as sand, clay or volcanic ash. It can also be made of other natural organic or synthetic materials, which need to be pre-treated with a buffer solution.

[0056] Absorbent pad (42): Vertically fixed in the horizontal direction at the other end (37) of the nitrocellulose membrane (35).

[0057] The components of each biosensor (28) are fixed on the inner surface of the outer plate (29) of the biosensor unit (27) using an adhesive. At the same time, the components of each biosensor (28) are tightly connected to the base membrane (31) to ensure that samples and reagents can migrate smoothly on the biosensor (28) (also known as the test strip) during the test. The overlapping part between the components is preferably about 2 mm.

[0058] Cerumen, or earwax, diffuses into the sample pad (41), and the sample pad acts as a filter to facilitate the flow of samples and reagents. The conjugate (enzyme or antibody) is added dropwise to the conjugate pad (40) using a pipette. In other modes, the cushion layer can be prefabricated separately, dried to room temperature, stored in a dryer at 4 °C, and then combined with the biosensor when in use. The corresponding capture reagent is distributed in the test line (38) and control line (39) areas of the nitrocellulose membrane (35) through appropriate equipment or methods.

[0059] The marker (tracer) is distributed on the cushion layer and the nitrocellulose membrane, and the preferred components are colloidal gold, carbon or latex. The specific marker is selected according to the compatibility with the reagent components.

[0060] Once the specific analyte in the cerumen reaches the conjugate pad (40), the conjugated tracer antibody will be rehydrated and bind to the analyte (the area A marked in Figure 10).

[0061] The formed complex moves from area A to area D by capillary action and is detected at the pre-coated antibody (area B) on the test line (38). If no complex is produced on the test line (38), it shows a negative result.

[0062] The staining results that each biosensor (28) may show are: Figure 11a Showing a positive result; Figure 11b Showing a negative result; Figure 11c Showing an invalid result.

[0063] On the test line (38), the colloidal gold-labeled antibody recognizes and binds to the unique part of the target molecule, namely the antigenic epitope.

[0064] Unreacted labeled antibody or labeled substance is finally captured by the specific antibody (region C) on the control line (39). Both the test line (38) and the control line (39) contain capture reagents to display the test results.

[0065] The control line (39) shows color, indicating that the test has been correctly performed. The label is responsible for showing color on the control line (39) and the test line (38).

[0066] Finally, the absorption pad (42) is responsible for collecting any excess sample and reagent (region D).

[0067] During the test, one or more (preferably up to four) cylindrical multiplex lateral flow biosensor units (27) with cerumen samples pre-prepared are inserted into the biosensor holder (17), which is then inserted into the central circular opening (16) of the biomarker measuring device, making the upper outer shell (25) of the holder contact the top outer shell (11) of the measuring device to position it. After the holder (17) is inserted into the measuring device, the cameras (19a, 19b, 19c, 19d) inside the outer plate (29) of the biosensor unit (27) around the strut (18) are all aligned with the positions of the test line (38) and the control line (39), that is, the test strip or the stained area of the multiplex lateral flow biosensor.

[0068] Each camera (19a, 19b, 19c, 19d) captures images of the test line (38) and the control line (39) (stained area), generating dot matrix data arranged horizontally and vertically. Each point, or pixel, is represented by three numerical parameters corresponding to the red, green, and blue (RGB) channels of the image, forming an RGB circuit. The numerical value recorded by each channel will increase correspondingly with the increase in staining intensity.

[0069] The images obtained by each camera (19a, 19b, 19c, 19d) are processed in the information processing unit by immunochromatography software, which generates four sets of readings corresponding to the chronic levels of the measured substances in the cerumen sample. The results are displayed on the screen (P), and each screen corresponds to a camera (19a, 19b, 19c, 19d) and its associated biosensor (28). In other modes, the information processing unit software responsible for processing the information of the cameras (19a, 19b, 19c, 19d) can include artificial intelligence algorithms, which are trained to better interpret the images obtained by the cameras, thereby improving the accuracy of biomarker measurement.

[0070] Kit usage

[0071] Before use, the user needs to prepare the multiplex lateral flow biosensor (27) according to the following steps:

[0072] · Coat the conjugate (enzyme or antibody) to be used on the conjugate pad (40).

[0073] · Coat the corresponding capture reagent on the test line (38) and control line (39) of the nitrocellulose membrane (35).

[0074] Position.

[0075] Insert the cylindrical biosensor unit (27) into the biosensor holder (17), ensuring that the limiting device (30) on the outer plate (29) of the biosensor unit snaps into the groove (26) of the holder, thereby fixing the biosensor unit (27) within the holder (17).

[0076] Sample extraction should preferably use a sample extraction device similar to that described in WO2019 / 123392A1, the characteristics of which have been described above. However, if the sample extraction device is equipped with a sponge that can adapt to the shape and size of the cylindrical opening after the biosensor unit (27) is inserted into the holder (17), and the length is sufficient to allow sufficient cerumen sample to contact the sample pad (41) of each biosensor (28), then other devices can also be used for sample extraction. In the preferred mode, cortisol and glucose can be analyzed from a cerumen sample of only 0.8 mg. Therefore, the recommended minimum sample volume is 0.8 mg.

[0077] When collecting cerumen, insert the tip of the sample extraction device equipped with the sponge (8) into the ear canal and rotate the sponge in the ear canal for 30 to 60 seconds.

[0078] After obtaining the cerumen sample, the user needs to insert the sponge tip of the sample extraction device into the biosensor unit (27) that has been placed in the biosensor holder, as Figure 12 shown.

[0079] Inside the biosensor unit (27), the user needs to rotate the sponge (8) so that it contacts the sample pad (41) of each biosensor (28), distributing the cerumen biological sample over each sample pad (41) (the recommended minimum sample volume is 0.8 mg).

[0080] When the sample pad (41) is infiltrated with the cerumen sample, insert the biosensor unit (27) into the central circular opening (16) of the biomarker measurement device, making the upper housing (25) of the holder contact the top housing (11) of the measurement device for positioning, as Figure 13 shown.

[0081] To ensure that each control line (38) and test line (39) is aligned with the corresponding camera (19a, 19b, 19c, 19d), the biosensor holder (17) can be equipped with a guiding device (not shown in the figure) for use in conjunction with the central circular opening (16) of the housing.

[0082] As described above, the biomarker measurement device interprets various electrochemical signals as a mechanism for reading different biomarkers, such as glucose in human cerumen. However, other optical reading mechanisms can also be employed.

[0083] The change in fluorescence intensity can identify the target analyte, such as the electrical signal and specific concentration of glucose, through a specific calibration curve stored in the device's memory. Similarly, other conjugates, such as gold or silver nanoparticles of different wavelengths, can also be used to identify different concentrations.

[0084] In other modes, the biomarker measurement device of the present invention can also use other types of biosensors, including but not limited to: amperometric sensors, potentiometric sensors, impedance sensors, voltammetric sensors, piezoelectric sensors, temperature sensors, and optical sensors, etc.

[0085] Some biosensors may not require a camera because the measurement results can be sent directly or wirelessly to the information processing unit as signals. In this mode, there is no need for a camera support, lighting device, or focusing device. In addition, such biosensors may not require any pretreatment by the user (such as coating conjugates or capture reagents).

[0086] Similarly, in other modes, the cross-section of the sponge (8) of the sample extraction device, the central opening (16) of the biomarker measurement device, the biosensor carrier (17), and the biosensor unit (27) can be of any shape, as long as it can ensure that the sponge (8) can distribute an appropriate amount of cerumen sample to the biosensor. Even the biosensor carrier can only include structures and devices for fixing one or more sensors.

[0087] Similarly, the cross-section of the housing (10) of the biomarker measurement device can also be of any shape, such as circular, or only include structures and devices for fixing the biosensor carrier (17).

[0088] Finally, in other modes, the biosensor carrier (17) can also be not used, and the biosensor unit (27) can be directly inserted into the biomarker measurement device, provided that the biosensor unit (27) must have a cavity capable of inserting the sponge of the sample extraction device so that the sensor can be wetted by the cerumen sample.

[0089] Finally, it should be noted that the kits and methods for self-collection of cerumen and in-situ biomarker measurement of the present invention are not limited to the modes described above. Experts in the relevant field can make changes to the kits and methods for self-collection of cerumen and in-situ biomarker measurement of the present invention according to the guidance provided herein, and the changes can only be made within the scope of the following permission regulations.

Claims

1. A device for measuring biomarkers in a cerumen sample, which obtains the sample through a sample extraction device. The sample extraction device includes a handle and a head connected to the handle. The head contains a sponge bracket and a sponge fixed on the bracket. The cerumen sample is collected by inserting the sponge into the ear canal. The biomarker measuring device includes: A housing: at least including a top housing with an opening in the center; An information processing unit installed inside the housing; At least one display screen for displaying information. Each display screen is connected to the information processing unit to display the biomarker measurement results; One or more biosensors, each sensor is installed inside the housing and can contact the sponge of the sample extraction device. The cerumen sample on the sponge can be transferred and distributed to each biosensor.

2. The biomarker measuring device according to claim 1 may additionally include: A bottom housing: A pillar for supporting a camera, fixed in the central area of the surface of the bottom housing; At least one camera installed on the top of the pillar. Each camera should be positioned to align with a corresponding biosensor and connected to the information processing unit; A focusing device surrounding the pillar and one or more cameras to improve the resolution of the images obtained by the cameras; A light source located inside the housing.

3. The biomarker measuring device according to claim 2, wherein the focusing device includes a circular lens surrounding the camera pillar and the cylindrical biosensor, and is fixed by a support structure fixed on the bottom housing. The light source can generate monochromatic light with specific spectral characteristics in the wavelength range of 600nm to 1400nm.

4. The biomarker measuring device according to claim 1, wherein one or more biosensors can be connected to a detachable biosensor unit, and the unit includes: A hollow cylindrical flexible outer plate with both ends open, made of a material such as nitrocellulose that has a strong binding force with proteins such as antibodies or enzymes and different absorption characteristics (different capillary flow times). The cylindrical outer plate has an outer surface and an inner surface, and each biosensor is fixed on the inner surface of the outer plate; The biosensor unit is inserted into the biomarker measuring device through the central opening.

5. The biomarker measuring device according to claim 1, wherein the central opening of the housing is cylindrical, and the biomarker measuring device further additionally includes: A biosensor bracket, which is a hollow cylindrical structure. Its cylindrical housing includes an outer surface, an inner surface, an upper end and a lower end. The diameter of the upper end of the housing is larger than that of the cylindrical main body, and the lower end is open. The diameter of the cylinder can just tightly pass through the central circular opening of the biomarker measuring device; One or more biosensors are connected to a biosensor unit, which includes a hollow cylindrical flexible outer plate that is open at both ends and made of a material with strong binding force to proteins such as antibodies or enzymes, such as nitrocellulose. The cylindrical outer plate has an outer surface and an inner surface, with appropriate diameter and height, and can just be tightly inserted into the biosensor holder. Each biosensor is fixed on the inner surface of the outer plate. The biosensor unit is inserted into the inside of the biosensor holder, and the holder is inserted into the central opening of the cylindrical housing.

6. The biomarker measuring device according to claim 1, wherein the central opening of the housing is cylindrical, and the biomarker measuring device further includes: Bottom housing: A pillar for supporting the camera, fixed in the central area of the surface of the bottom housing; At least one camera, mounted on the top of the pillar. Each camera should be positioned to align with a corresponding biosensor and connected to the information processing unit; A focusing device, surrounding the pillar and one or more cameras to improve the resolution of the images obtained by the cameras; A light source located inside the housing, A biosensor holder, which is a hollow cylindrical structure. Its cylindrical housing includes an outer surface, an inner surface, an upper end and a lower end. The diameter of the upper end of the housing is larger than that of the cylindrical body, and the lower end is open. The diameter of the cylinder can just tightly pass through the central circular opening of the biomarker measuring device. One or more biosensors are connected to a biosensor unit, which includes a hollow cylindrical flexible outer plate that is open at both ends and made of a material with strong binding force to proteins such as antibodies or enzymes, such as nitrocellulose. The cylindrical outer plate has an outer surface and an inner surface, with appropriate diameter and height, and can just be tightly inserted into the biosensor holder. The biosensor unit is inserted into the inside of the biosensor holder, and the holder is inserted into the central opening of the cylindrical housing; Wherein: Each biosensor is equipped with a reaction test strip (staining area); The upper housing of the biosensor holder contacts the top housing (11) of the measuring device to limit its position; When the biosensor holder is inserted into the biomarker measuring device, the inner wall of the outer plate of the biosensor unit surrounds the pillar with the camera, so that each camera is aligned with the reaction test strip (staining area) of a multiplex lateral flow biosensor; The images obtained by each camera are processed by the immunochromatography software in the information processing unit. The software tests four readings, corresponding to the chronic levels of the substances measured in the cerumen, and the results are displayed on the screen. Each screen corresponds to a camera and its corresponding biosensor.

7. The biomarker measuring device according to claim 6, wherein each sensor is a multiplex lateral flow biosensor, including: An elongated base film, made of a flexible material, which may include but is not limited to vinyl polymers or nitrocellulose. The base film is pressure-sensitive and is fixed by a stable adhesive made of acrylic, without chemical reaction with the adhesive. The base film includes one end, the other end, an inner surface and an outer surface. The inner surface has hydrophilic properties and can slow down the diffusion of hydrophobic cerumen samples, thus ensuring the time required for immunohistochemical reactions; An elongated nitrocellulose film, including one end, the other end, a test line area and a control line area. The nitrocellulose film is vertically fixed on the base film and extends along its entire length. Its one end and the other end are flush with one end and the other end of the base film respectively; A conjugate pad, vertically fixed in the horizontal direction at one end of the nitrocellulose film, for accommodating conjugated labeled antibodies; A cerumen sample pad, vertically fixed above the conjugate pad. The material of the sample pad has a high absorption capacity for grease and can be natural inorganic materials such as sand, clay or volcanic ash, or can also be made of other natural organic or synthetic materials, and needs to be pre-treated with a buffer solution; An absorbent pad, vertically fixed in the horizontal direction at the other end of the nitrocellulose film; Wherein: The cerumen diffuses to the sample pad, and the sample pad also acts as a filter; The conjugate (enzyme or antibody) used is coated on the conjugate pad; The corresponding capture reagents are coated on the test line and control line areas of the nitrocellulose film; The label (tracer) is distributed on the pads and the nitrocellulose film, and the components can be but are not limited to colloidal gold, carbon or latex; The complex formed during the test is displaced by capillary action and is detected by the detection antibody pre-coated in the test line area.

8. Each kit for self-collection of cerumen and in-situ biomarker measurement includes: A sample extraction device, including a handle and a head connected to the handle. The head includes a bracket for fixing a sponge and a sponge fixed on the bracket, and can be inserted into the ear canal to collect cerumen samples; A biomarker measurement device, for measuring the biomarkers in the cerumen samples collected by the sample extraction device described in claims 1 to 7.

9. A method for self-collection of cerumen and in-situ biomarker measurement includes the following steps: a. Collect cerumen samples through an extraction device, which includes a handle and a head connected to the handle. The head includes a bracket for fixing a sponge and a sponge fixed on the bracket, and can be inserted into the ear canal to collect cerumen samples; b. Transfer the cerumen samples from the sponge of the extraction device to one or more biosensors in the biomarker measurement device described in claims 1 to 7.

10. A multiplex lateral flow biosensor comprises: An elongated base film, made of a flexible material, which may include but is not limited to vinyl polymers or nitrocellulose. The base film is pressure-sensitive and is fixed by a stable adhesive made of acrylic, without chemical reaction with the adhesive. The base film includes one end, the other end, an inner surface and an outer surface. The inner surface has hydrophilic properties and can slow down the diffusion of hydrophobic cerumen samples, thus ensuring the time required for immunohistochemical reactions; An elongated nitrocellulose membrane, including one end, the other end, a test line area, and a control line area, is vertically fixed above the base membrane and extends along its entire length, with one end and the other end flush with one end and the other end of the base membrane respectively; A conjugate pad is vertically fixed in the horizontal direction at one end of the nitrocellulose membrane for accommodating a conjugated tracer antibody; A cerumen sample pad is vertically fixed above the conjugate pad. The material of the sample pad has a high absorption capacity for grease and can be a natural inorganic material such as sand, clay, or volcanic ash, or can be made of other natural organic or synthetic materials and needs to be pre-treated with a buffer solution; An absorbent pad is vertically fixed in the horizontal direction at the other end of the nitrocellulose membrane; Wherein: Cerumen diffuses to the sample pad, and the sample pad also functions as a filter; The conjugate (enzyme or antibody) used is coated on the conjugate pad; The corresponding capture reagents are coated on the test line and control line areas of the nitrocellulose membrane; The label (tracer) is distributed on the pads and the nitrocellulose membrane, and the components can be but are not limited to colloidal gold, carbon, or latex; The complex formed during the test is displaced by capillary action and detected by the pre-coated detection antibody in the test line area.

Images