A multilayer composite metapneumovirus detection reagent strip

Abstract

The utility model belongs to virus detection reagent strip technical field discloses a multilayer composite metapneumovirus detection reagent strip, including filter sample pad, virus enrichment pad, signal amplification pad, chromatography pad and water absorption pad which are stacked in turn along sample flow direction, virus enrichment pad is provided with virus capture area, the top of one end of virus enrichment pad far from virus capture area is provided with solid-state elution salt block, and buffer pad is arranged between solid-state elution salt block and virus enrichment pad, the utility model discloses filter sample pad, virus enrichment pad, signal amplification pad and chromatography pad can remove impurity in advance, ensure the specificity of virus capture area and the purity of detection line signal, thereby significantly reduce the incidence of false positive and false negative result, and then sample enters virus capture area and is captured and fixed, forms local high concentration virus enrichment, then after elution enters signal amplification pad and chromatography pad, greatly enhances the detectability of low concentration virus.

Description

Technical Field

[0001] This utility model relates to the field of virus detection reagent strip technology, and in particular to a multi-layer composite metapneumovirus detection reagent strip. Background Technology

[0002] Human metapneumovirus (HMV) is one of the important pathogens causing acute respiratory infections (such as bronchiolitis and pneumonia) in infants, the elderly, and immunocompromised individuals. Its clinical symptoms are similar to those of respiratory syncytial virus (RSV). Therefore, rapid and accurate laboratory diagnosis is crucial for clinical differential diagnosis, rational drug use, and infection control.

[0003] Currently, clinical testing methods for hMPV mainly fall into two categories: viral nucleic acid detection (such as PCR) and viral antigen detection. Although PCR has high sensitivity and specificity, the testing process is cumbersome and time-consuming, requiring specialized equipment and operators, making it unsuitable for primary healthcare institutions or bedside testing scenarios that require rapid diagnostic results.

[0004] Therefore, immunochromatography is widely used as a rapid detection technique. This type of technique typically uses a nitrocellulose membrane as a carrier, allowing the sample solution to flow along the chromatography strip via capillary action. The specific binding of labeled antibodies to viral antigens results in color development at the test line (T line) and control line (C line) for interpretation. However, existing hMPV immunochromatographic test strips still have significant limitations in practical applications: Significant interference from sample impurities: Respiratory samples (such as nasopharyngeal swab wash) typically contain large amounts of mucus, cell debris, and other non-specific proteins. These impurities easily clog the pores of the chromatography membrane, hindering liquid flow and leading to chromatography failure (false negatives); or they may bind to non-specific antibodies, producing background noise or even false positive results, severely affecting the specificity and accuracy of the test.

[0005] Insufficient detection sensitivity: Especially in the early stages of infection or in pediatric patients, viral load in samples is usually low. In traditional one-step immunochromatographic techniques, the binding time between viral antigen and labeled antibody is short, and the antigen is diluted during flow, making it difficult to accumulate sufficient signal intensity at the T line. This results in a high rate of false negatives for low-concentration samples, failing to meet the needs of early diagnosis.

[0006] To address this, a multi-layer composite metapneumovirus detection strip is proposed. Utility Model Content

[0007] The purpose of this invention is to provide a multi-layer composite metapneumovirus detection reagent strip, thereby solving or at least alleviating one or more of the above-mentioned problems and other problems existing in the prior art.

[0008] To achieve the above objectives, the main technical solutions adopted by this utility model include: A multilayer composite metapneumovirus detection strip includes a filter sample pad, a virus enrichment pad, a signal amplification pad, a chromatography pad, and an absorbent pad stacked sequentially along the sample flow direction. The virus enrichment pad is provided with a virus capture area for concentrated capture of metapneumovirus. The virus capture area is located at one end of the virus enrichment pad near the signal amplification pad. A solid elution salt block is disposed above the end of the virus enrichment pad away from the virus capture area. A slow-absorption pad is disposed between the solid elution salt block and the virus enrichment pad.

[0009] In a multi-layer composite metapneumovirus detection reagent strip according to the present invention, the absorbent pad is a dense cellulose filter paper pad.

[0010] In a multi-layer composite metapneumovirus detection reagent strip according to the present invention, the bottom of the absorbent pad is provided with several arched channels equidistantly distributed along the width direction of the virus capture area.

[0011] In a multi-layer composite metapneumovirus detection reagent strip according to the present invention, the solid elution salt block is a dried citrate block.

[0012] In a multilayer composite metapneumovirus detection reagent strip according to the present invention, the virus enrichment pad is a nitrocellulose membrane, and the virus capture area is coated with metapneumovirus polyclonal antibodies.

[0013] According to the present invention, a multilayer composite metapneumovirus detection reagent strip is provided, wherein the signal amplification pad is a glass fiber membrane, and the signal amplification pad is coated with colloidal gold-labeled metapneumovirus monoclonal antibody and colloidal gold-labeled mouse IgG antibody.

[0014] In a multi-layer composite metapneumovirus detection reagent strip according to the present invention, the filter sample pad is a glass fiber pad, and the surface of the filter sample pad is modified with a cellulose coating for degrading mucus impurities in the sample.

[0015] According to the present invention, a multilayer composite metapneumovirus detection reagent strip is provided, wherein the chromatography pad is a nitrocellulose membrane, and a detection line and a control line are sequentially arranged on the chromatography pad along the sample flow direction. The detection line is coated with metapneumovirus polyclonal antibody, and the control line is coated with goat anti-mouse IgG antibody.

[0016] In a multi-layer composite metapneumovirus detection reagent strip according to the present invention, the absorbent pad is an absorbent cotton pad.

[0017] According to the present invention, a multi-layer composite metapneumovirus detection reagent strip further includes a shell, on which a sample application hole and an observation hole are provided, and a mounting groove is also provided on the shell, in which a magnifying lens is installed.

[0018] This utility model has at least the following beneficial effects: The sample first flows through the virus capture zone on the virus enrichment pad. The polyclonal antibody coated in this zone specifically captures and spatially concentrates the dispersed virus particles in the sample, achieving effective target enrichment. Subsequently, the eluent generated by the slow-release pad elutes the enriched virus from the capture zone, allowing it to enter the signal amplification pad at a locally high concentration and fully bind to the colloidal gold-labeled monoclonal antibody against metapneumovirus. This "enrichment-elution" process, combined with the signal amplification effect of colloidal gold, greatly enhances the detectability of low-concentration viruses and lowers the detection limit.

[0019] The cellulose coating on the surface of the filter sample pad can degrade mucus in the sample, reduce sample viscosity, prevent non-specific substances from clogging the membrane pores, and remove impurities in advance, reducing the possibility of non-specific binding in subsequent reactions. This ensures specific capture of the virus capture zone and purity of the detection line signal, thereby significantly reducing the incidence of false positive and false negative results.

[0020] The observation hole and magnifying lens on the housing help users observe the color development of the test line and control line more clearly. In particular, for weak positive signals, the magnification function can prevent missed detection and improve the accuracy of result interpretation. Attached Figure Description

[0021] The accompanying drawings, which are included to provide a further understanding of this application and form part of this application, illustrate exemplary embodiments and are used to explain this application, but do not constitute an undue limitation of this application. In the drawings: Figure 1 This is a schematic diagram of the model of this utility model; Figure 2 This is a schematic diagram of the structure of this utility model; Figure 3 This is a schematic diagram of the structure of the absorbent pad of this utility model; Figure 4 This is a schematic diagram of the structure of this utility model after the housing is installed; Figure 5 This is a schematic diagram of the exploded structure of this utility model.

[0022] Explanation of icon numbers: 1. Sample filter pad; 2. Virus enrichment pad; 201. Virus capture zone; 202. Solid elution salt block; 203. Slow-release pad; 2031. Arched channel; 3. Signal amplification pad; 4. Chromatography pad; 401. Detection line; 402. Quality control line; 5. Absorbent pad; 6. Base plate; 7. Shell; 701. Sample dispensing port; 702. Observation port; 703. Mounting slot; 704. Magnifying lens. Detailed Implementation

[0023] The following will describe in detail the implementation of this application with reference to the accompanying drawings and embodiments, so that the implementation process of how this application uses technical means to solve technical problems and achieve technical effects can be fully understood and implemented accordingly.

[0024] Please refer to Figures 1 to 5 As shown, an embodiment of this utility model provides a multilayer composite metapneumovirus detection reagent strip, including a filter sample pad 1, a virus enrichment pad 2, a signal amplification pad 3, a chromatography pad 4, and an absorbent pad 5 stacked sequentially along the sample flow direction. All pads are fixed on a base plate 6, and the overlap width of adjacent pads is 1-2 mm (to ensure smooth sample chromatography and avoid strip breakage). The virus enrichment pad 2 is provided with a virus capture area 201 for concentrated capture of metapneumovirus. The virus capture area 201 is located at the end of the virus enrichment pad 2 near the signal amplification pad 3. A solid elution salt block 202 is provided above the end of the virus enrichment pad 2 away from the virus capture area 201. A slow-absorption pad 203 is provided between the solid elution salt block 202 and the virus enrichment pad 2.

[0025] By adopting the above technical solution, a multi-layer detection system with time-series control function was constructed. The functions of each pad in the sample flow direction are clearly defined: the filter sample pad 1 is responsible for the initial screening of impurities; the core area of ​​the virus enrichment pad 2, the virus capture zone 201, is responsible for the specific capture and enrichment of the target virus; the combination of the upper slow-absorption pad 203 and the solid elution salt block 202 constitutes a delayed-start elution solution release system, providing a structural basis for the subsequent sensitive detection process of "capture first, then elute".

[0026] In this embodiment, the bottom of the absorbent pad 203 is provided with several arched channels 2031 that are equidistantly distributed along the width direction of the virus capture area 201. The number of arched channels 2031 is 3-5, the width of a single channel is 0.5-0.8mm, the height is 0.3-0.5mm, and the channel spacing is 1-1.5mm. This size design can ensure that the sample passes through quickly while avoiding the channel being too wide, which would reduce the water absorption efficiency of the absorbent pad 203.

[0027] By adopting the above technical solution, the arched channel 2031 provides a direct path for the sample liquid flow to bypass the dense absorbent pad 203 in the early stage of detection, ensuring that the sample can flow quickly and fully to and saturate the virus capture area 201, ensuring the virus capture efficiency, while avoiding the absorbent pad 203 absorbing water too early and affecting subsequent functions.

[0028] In this embodiment, the slow-absorbing pad 203 is a dense cellulose filter paper pad with a pore size of 5-10 μm and a thickness of 0.6-0.8 mm. Its water absorption rate is controlled at 0.1-0.2 mL / min (at 25°C). This dense structure can precisely control the liquid absorption rate and avoid the eluent from being generated too early or too late.

[0029] By adopting the above technical solution, a dense cellulose filter paper pad is used as a slow-absorbing pad 203. Its microstructure can effectively control the lateral diffusion and flow rate of the liquid, ensuring that it can slowly absorb the liquid and dissolve the solid elution salt block 202 above, generating a uniform and gentle elution liquid flow, thus achieving controllable elution.

[0030] In this embodiment, the solid elution salt block 202 is a dried citrate block with a mass of 5-8 mg and a purity of ≥99%. It is formed by freeze-drying (to avoid moisture absorption and clumping of the salt block and to ensure uniform concentration after dissolution). The pH of the eluent formed after dissolution is stable at 6.0-6.5. This pH range can effectively destroy the binding bond between the virus and the antibody without affecting the activity of the colloidal gold-labeled antibody.

[0031] By adopting the above technical solution, dry citrate blocks are used as solid elution salt blocks 202. After being wetted by the absorbent pad 203, they dissolve and provide a stable pH and ionic strength environment, effectively disrupting the binding between the virus and the antibody in the virus capture zone 201, and efficiently eluting the enriched virus.

[0032] In this embodiment, the virus enrichment pad 2 is a nitrocellulose membrane. The virus capture zone 201 is coated with a polyclonal antibody against metapneumovirus. The polyclonal antibody is immobilized in the virus capture zone 201. The polyclonal antibody is derived from the anti-hMPV whole virus antibody prepared by immunizing New Zealand white rabbits. After purification with Protein A, the coating concentration is 1.5-2.5 μg / cm². After coating, it is dried at 37°C for 4-6 hours (to ensure that the antibody is firmly bound to the membrane and to avoid detachment during chromatography).

[0033] By adopting the above technical solution, the virus enrichment pad 2 made of nitrocellulose membrane has good protein binding ability. The virus capture area 201 near the signal amplification pad 3 is coated with polyclonal antibodies against metapneumovirus. Taking advantage of the rich binding epitopes of polyclonal antibodies, the virus can be captured extensively and efficiently, achieving local enrichment of the virus.

[0034] In this embodiment, the signal amplification pad 3 is a glass fiber membrane, coated with colloidal gold-labeled metapneumovirus monoclonal antibody and colloidal gold-labeled mouse IgG antibody. The colloidal gold particles have a particle size of 30-40 nm (prepared by the trisodium citrate reduction method, with a particle size variation coefficient ≤10%). The metapneumovirus monoclonal antibody is an anti-hMPVF protein-specific monoclonal antibody (recognizing the 230-250 amino acid epitope of the F protein), with a labeling concentration of 0.8-1.2 mg / mL. The mouse IgG antibody is a conventional myeloma cell secretion antibody, with a labeling concentration of 0.5-0.8 mg / mL. The two antibodies are mixed at a volume ratio of 1:1 and then uniformly sprayed onto the glass fiber membrane using a spraying device at a spraying volume of 2-3 μL / cm. After spraying, the membrane is vacuum-dried at 30°C for 2 hours (to ensure that the antibody is dry and stable without loss of activity).

[0035] By employing the above technical solution, the signal amplification pad 3, composed of a glass fiber membrane, can stably adsorb dried labeled antibodies. The colloidal gold-labeled metapneumovirus monoclonal antibody coated on it binds to the eluted virus to form a detection complex; the colloidal gold-labeled mouse IgG antibody serves as a reference, flowing to the control line 402 where it is captured to verify the effective completion of the detection process.

[0036] In this embodiment, the filter sample pad 1 is a glass fiber pad. The surface of the filter sample pad 1 is modified with a cellulose coating for degrading mucus impurities in the sample. The pore size of the glass fiber pad is 10-20 μm and the thickness is 0.8-1.2 mm. The cellulose coating uses sodium carboxymethyl cellulose (CMC-Na) with a modification concentration of 0.5-1.0 mg / cm². After modification, it is dried in a forced-air drying oven at 50°C for 3 hours (to ensure that the coating adheres firmly and avoids falling off and contaminating the sample during chromatography). This coating can degrade more than 80% of the mucin in the sample within 5 minutes, significantly reducing the sample viscosity.

[0037] By adopting the above technical solution, the glass fiber pad itself has good filtration capacity, and the cellulose coating modified on its surface can further decompose mucus impurities such as mucin commonly found in respiratory samples, reduce sample viscosity, reduce non-specific interference, and improve chromatography efficiency and detection specificity.

[0038] In this embodiment, the chromatography pad 4 is a nitrocellulose membrane. A detection line 401 and a control line 402 are sequentially arranged on the chromatography pad 4 along the sample flow direction. The detection line 401 is coated with a polyclonal antibody against metapneumovirus, and the control line 402 is coated with goat anti-mouse IgG antibody. The pore size of the nitrocellulose membrane is 8-12 μm (to ensure that the "virus-colloidal gold complex" can pass through smoothly without non-specific adsorption). The coating concentration of the detection line 401 is 2.0-3.0 μg / cm², and the coating concentration of the control line 402 is 1.0-1.5 mg / cm². The line width of both lines is 1.0-1.2 mm, and the spacing is 3-4 mm. After coating, the membrane is dried at 37°C for 8 hours and then sealed for storage (to prevent the antibody from becoming damp and ineffective).

[0039] By employing the above technical solution, a test line 401 and a control line 402 are set on the chromatography pad 4 of the nitrocellulose membrane. The test line 401, coated with a polyclonal antibody against metapneumovirus, is used to capture the "virus-colloidal gold-labeled monoclonal antibody" complex, showing a positive result. The control line 402, coated with goat anti-mouse IgG antibody, is used to capture free colloidal gold-labeled mouse IgG antibody. This line should show color regardless of whether the sample contains the virus, serving as an internal process control.

[0040] In this embodiment, the absorbent pad 5 is an absorbent cotton pad made of degreased cotton, with a thickness of 1.5-2.0 mm, a width of 8-10 mm, and an absorbency of ≥1.5 mL (to ensure complete absorption of the sample liquid throughout the chromatography process and avoid liquid residue leading to misjudgment of results); the end of the absorbent pad 5 extends 2-3 mm beyond the chromatography pad 4 to ensure sufficient capillary traction.

[0041] By adopting the above technical solution, an absorbent cotton pad is used as the absorbent pad 5. Its strong water absorption provides a continuous capillary traction force for the entire chromatography process, ensuring that the liquid can flow smoothly and continuously through all functional pad layers until the detection is completed.

[0042] In this embodiment, a housing 7 is also included. The housing 7 has a sample application hole 701 and an observation hole 702. The housing 7 also has a mounting groove 703, in which a magnifying lens 704 is installed. The housing 7 is made of polypropylene (temperature resistance range -20℃ to 60℃, to avoid deformation of the housing due to high or low temperatures). The diameter of the sample application hole 701 is 3-4mm (matching the size of conventional pipette tips for easy sample application). The size of the observation hole 702 is 8mm×15mm (completely covering the detection line and quality control line area). The magnifying lens 704 is made of acrylic and has a magnification of 1.5-2.0x (clearly magnifying the colorimetric signal without causing distortion of the field of view). The edge of the lens is fixed to the mounting groove 703 by ultrasonic welding (to prevent the lens from falling off during use).

[0043] By adopting the above technical solution, the housing 7 provides physical protection for the internal reagent strip. The sample dispensing port 701 guides the sample to be accurately dispensed onto the filter sample pad 1; the observation port 702 allows the user to observe the results of the detection line 401 and the control line 402; the magnifying lens 704 can magnify and display the chromatography results, making it easier to interpret weak signals and improving the convenience of detection and the accuracy of result interpretation.

[0044] Working principle: 1. Sample Loading and Preliminary Filtration: The liquid sample must be nasopharyngeal swab eluate (the sample should be pre-equilibrated at room temperature (20-25℃) for 5 minutes to avoid abnormal chromatography speed due to excessively low temperature). Take 100-150 μL of sample and slowly drop it onto the filter sample pad 1 through the sample application port 701 on the housing 7. Avoid generating air bubbles during the drop process (to prevent air bubbles from clogging the chromatography channels). The cellulose coating on the surface of the filter sample pad 1 degrades the mucus impurities in the sample within 3-5 minutes. The preliminarily purified sample flows forward at a speed of 0.3-0.5 mm / min under capillary action.

[0045] 2. Virus Capture and Enrichment: The preliminarily purified sample rapidly reaches the virus capture area 201 of the virus enrichment pad 2 within 2-3 minutes through the arched channel 2031 at the bottom of the absorbent pad 203. At this point, it needs to be left to stand at room temperature (20-25℃, relative humidity 40%-60%) for 3-5 minutes to ensure the sample fully infiltrates the virus capture area 201. The polyclonal antibody against metapneumovirus in the area specifically captures and enriches the metapneumovirus in the sample (capture efficiency ≥95%), and fixes the virus in the virus capture area 201.

[0046] 3. Controlled Elution and Signal Labeling: As the liquid continues to flow, the absorbent pad 203 absorbs the sample liquid from its surroundings. After approximately 5-8 minutes, it begins to gradually dissolve the solid citrate block 202 above, generating a citrate eluent with a concentration of 50-100 mmol / L and a pH of 6.0-6.5. The eluent flows through the virus capture zone 201 at a speed of 0.5-1 mm / min. At this flow rate, the eluent can efficiently disrupt the binding bonds between the virus and the antibody (elution efficiency ≥90%), eluting the enriched virus. The virus-laden eluent then enters the signal amplification pad 3, where it binds to the colloidal gold-labeled metapneumovirus monoclonal antibody within the pad, forming a "virus-colloidal gold-labeled monoclonal antibody" complex (binding time approximately 2-3 minutes). Simultaneously, free colloidal gold-labeled mouse IgG antibodies within the signal amplification pad 3 move synchronously with the liquid flow.

[0047] 4. Chromatographic Detection and Result Interpretation: Under the capillary traction of the absorbent pad 5, the above-mentioned complex and free labeled antibody continue to be chromatographically separated to the chromatography pad 4 at a speed of 0.2-0.4 mm / min. First, the "virus-colloidal gold-labeled monoclonal antibody" complex is specifically captured by the polyclonal antibody coated on the detection line 401. Colloidal gold particles aggregate at the detection line, forming a visible red band after about 10-15 minutes (the band width is positively correlated with the concentration; the band color is lighter in low-concentration samples and needs to be observed through a magnifying lens 704). Unbound free colloidal gold-labeled mouse IgG antibody continues to move forward and is captured by the goat anti-mouse IgG antibody on the control line 402, forming a red control band (if the control line does not develop color, it indicates an abnormal detection process, and the result is invalid). After the test is completed, the user interprets the results through the observation hole 702 with the assistance of the magnifying lens 704: if both the test line and the control line show color, it is positive (indicating that the sample contains hMPV); if only the control line shows color, it is negative (indicating that the sample does not contain hMPV); if the control line does not show color, the result is invalid regardless of whether the test line shows color (re-sampling and testing are required).

[0048] Performance verification and usage precautions: 1. Performance verification metrics: Detection limit: For the detection of hMPV virus solutions (concentrations of 1 PFU / mL, 5 PFU / mL, 10 PFU / mL, and 20 PFU / mL), the lowest detection limit of this test strip is 5 PFU / mL, which is 4 times more sensitive than the traditional immunochromatographic test strip (detection limit of 20 PFU / mL). Specificity: No false positives were observed in the detection of respiratory syncytial virus (RSV), influenza A / B virus, and adenovirus positive samples (no color development of the test line), with a specificity of 100%. Repeatability: For 10 parallel tests of hMPV positive samples at 10 PFU / mL, the coefficient of variation (CV) of the colorimetric intensity of the test line was 3.2%-5.8% (which meets the requirement of CV≤10% for clinical testing).

[0049] 2. Precautions for use: Storage conditions: The test strips should be stored at 2-8℃ in a dark and sealed environment. The shelf life is 12 months. Use within 1 hour after opening to avoid moisture, which may reduce the activity of the functional layer. Sample requirements: Only nasopharyngeal swab eluate samples are applicable. The samples must be tested within 2 hours of collection (if a delay is necessary, the samples must be refrigerated at 4°C and not exceed 24 hours). Environmental requirements: The test must be conducted in an environment of 20-25℃ and relative humidity of 40%-60% to avoid abnormal chromatography caused by high temperature and high humidity; Interpretation time: Results should be interpreted within 15-20 minutes after sample addition. If more than 20 minutes have passed, the colloidal gold may diffuse, leading to misinterpretation of the results.

[0050] The foregoing description illustrates and describes several preferred embodiments of the present invention. However, as previously stated, it should be understood that the present invention is not limited to the forms disclosed herein and should not be construed as excluding other embodiments. It can be used in various other combinations, modifications, and environments, and can be altered within the scope of the present invention's conception through the foregoing teachings or related technical or knowledge. Any modifications and variations made by those skilled in the art that do not depart from the spirit and scope of the present invention should be within the protection scope of the appended claims.

Claims

1. A multi-layer composite metapneumovirus detection strip, characterized in that, The sample includes a filter pad (1), a virus enrichment pad (2), a signal amplification pad (3), a chromatography pad (4), and an absorbent pad (5) stacked sequentially along the sample flow direction. The virus enrichment pad (2) is provided with a virus capture area (201) for concentrated capture of metapneumovirus. The virus capture area (201) is located at one end of the virus enrichment pad (2) near the signal amplification pad (3). A solid elution salt block (202) is provided above the end of the virus enrichment pad (2) away from the virus capture area (201). A slow-absorbing pad (203) is provided between the solid elution salt block (202) and the virus enrichment pad (2).

2. The multilayer composite metapneumovirus detection reagent strip according to claim 1, characterized in that: The bottom of the absorbent pad (203) has several arched channels (2031) that are equidistantly distributed along the width of the virus capture area (201).

3. The multilayer composite metapneumovirus detection reagent strip according to claim 2, characterized in that: The absorbent pad (203) is a dense cellulose filter paper pad.

4. The multilayer composite metapneumovirus detection reagent strip according to claim 3, characterized in that: The solid elution salt block (202) is a dried citrate block.

5. The multilayer composite metapneumovirus detection reagent strip according to claim 1, characterized in that: The virus enrichment pad (2) is a nitrocellulose membrane, and the virus capture zone (201) is coated with polyclonal antibodies against metapneumovirus.

6. The multilayer composite metapneumovirus detection reagent strip according to claim 1, characterized in that: The signal amplification pad (3) is a glass fiber membrane, and the signal amplification pad (3) is coated with colloidal gold-labeled metapneumovirus monoclonal antibody and colloidal gold-labeled mouse IgG antibody.

7. The multilayer composite metapneumovirus detection reagent strip according to claim 6, characterized in that: The filter sample pad (1) is a glass fiber pad, and the surface of the filter sample pad (1) is modified with a cellulose coating for degrading mucus impurities in the sample.

8. The multilayer composite metapneumovirus detection reagent strip according to claim 7, characterized in that: The chromatography pad (4) is a nitrocellulose membrane. A detection line (401) and a control line (402) are sequentially arranged on the chromatography pad (4) along the sample flow direction. The detection line (401) is coated with a polyclonal antibody against metapneumovirus, and the control line (402) is coated with a goat anti-mouse IgG antibody.

9. The multilayer composite metapneumovirus detection reagent strip according to claim 1, characterized in that: The absorbent pad (5) is an absorbent cotton pad.

10. A multilayer composite metapneumovirus detection strip according to any one of claims 1 to 9, characterized in that: It also includes a housing (7), on which a sample application hole (701) and an observation hole (702) are provided, and a mounting groove (703) is also provided on the housing (7), in which a magnifying lens (704) is installed.

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