Protein chip for detecting respiratory viruses

Abstract

The utility model discloses a protein chip for detecting respiratory viruses, which belongs to the technical field of biochips and diagnostic reagents and comprises an antigen loading membrane, a reverse osmosis membrane layer, a water absorption membrane layer and a leakage-proof membrane layer are sequentially fixed on the lower surface of the antigen loading membrane, and a detection area is arranged on the upper surface of the antigen loading membrane. The detection area comprises a plurality of antigen loading areas and a quality control area, a respiratory tract specific antigen is fixedly loaded on each antigen loading area, and the respiratory tract specific antigens comprise chlamydia pneumoniae, mycoplasma pneumoniae, adenovirus, syncytial virus, influenza B virus and parainfluenza virus. The percolation type protein chip can be used for detecting six pathogens related to the respiratory tract, and is simple in method and high in practicability.

Description

Technical Field

[0001] The utility model relates to the technical field of biochips and diagnostic reagents, in particular to a protein chip for detecting respiratory viruses. Background Art

[0002] Percolation biochips feature directional flow control, enabling control of reaction time and duration, thereby improving reaction efficiency. As disclosed in Chinese Patent CN 103439493 A, this chip comprises a chip housing with a window. Within the housing below the window, a nitrocellulose membrane, a reverse osmosis layer, a water-absorbing layer, and a leak-proof layer are arranged in order from top to bottom. Aptamer molecules targeting the target substance are immobilized on the surface of the nitrocellulose membrane.

[0003] This design avoids the problem of rapid liquid infiltration caused by direct contact between the chip matrix and the underlying water-absorbing material in traditional infiltration biochips, which in turn leads to uncontrollable reaction time and reaction process and reduced reaction efficiency.

[0004] In the existing technology, glass substrates are often used as antigen carriers for the detection of various respiratory viruses, as shown in Chinese patents CN 2901314Y and CN 114807438 A. However, during the production process of glass substrates, the glass surface needs to be modified by various treatments such as formaldehyde, isothiocyanate, and amination. The operation is complicated and the technical threshold is high. If the treatment is not done properly, it will lead to inaccurate test results.

[0005] In view of the above situation, the applicant selected a variety of common human respiratory viruses as antigens and used filtration biochip technology to detect these multiple viruses. The design method is simple and makes up for the shortcomings of existing technologies. Utility Model Content

[0006] The technical problem to be solved by the utility model is to design a new type of percolation protein chip for detecting 6 respiratory tract-related pathogens. The method is simple and practical.

[0007] In order to solve the above technical problems, the present invention adopts the following technical solutions: a protein chip for detecting respiratory viruses, comprising an antigen-loaded membrane, wherein a reverse osmosis membrane layer, a water-absorbing membrane layer and a leak-proof membrane layer are fixed in sequence on the lower surface of the antigen-loaded membrane, and a detection area is provided on the upper surface of the antigen-loaded membrane, wherein the detection area comprises several antigen-loading areas and a quality control area, and each antigen-loading area is fixedly loaded with a respiratory-specific antigen, wherein the respiratory-specific antigens include Chlamydia pneumoniae, Mycoplasma pneumoniae, adenovirus, syncytial virus, influenza B virus and parainfluenza virus.

[0008] Furthermore, at least two antigen loading areas are provided for each respiratory tract specific antigen.

[0009] Furthermore, the quality control area is immobilized with goat anti-mouse antibodies.

[0010] Furthermore, the quality control areas are respectively arranged at both ends of the antigen loading area.

[0011] Furthermore, the number of quality control areas at both ends of the antigen loading area is different.

[0012] Furthermore, two quality control areas are provided at the front end of the antigen loading area, and one quality control area is provided at the rear end.

[0013] Furthermore, the antigen loading membrane is arranged in a chip housing, and the chip housing comprises an upper housing and a lower housing, a window is arranged on the upper housing, and a column is arranged on the lower housing.

[0014] Furthermore, the antigen-loaded membrane is a nitrocellulose membrane, and the reverse osmosis membrane layer, the water-absorbing membrane layer and the leak-proof membrane layer are all made of cotton fiber structures. Beneficial effects

[0015] This application uses a percolation protein chip design to immobilize six common respiratory virus-specific antigens (CPN, MP, RSV, ADV, FLU B, and PIV) in a microarray format on a nitrocellulose membrane. Specific antibodies in the sample undergo a specific immune binding reaction with the antigen, forming an antigen-antibody immune complex on the nitrocellulose membrane. Colloidal gold-labeled mouse anti-human antibodies then specifically bind to the immune complex, ultimately resulting in visible red spots. The test results are then read using a biochip reader, enabling qualitative analysis of the sample.

[0016] Setting the quality control areas at both ends of the antigen loading area and setting different numbers of quality control areas can not only be used to distinguish the direction of the chip and position it, but also to determine whether the quality control reagent mouse anti-human antibody has deteriorated. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is a schematic diagram of the structure of the protein chip of the utility model.

[0018] Figure 2 This is a schematic diagram of the structure of the antigen loading area arrangement pattern of the utility model Figure 1 .

[0019] Figure 3 This is a schematic diagram of the structure of the antigen loading area arrangement pattern of the utility model Figure 2 .

[0020] Figure 4 This is a schematic diagram of the structure of the antigen loading area arrangement pattern of the utility model Figure 3 .

[0021] Figure 5 This is a schematic structural diagram of the chip housing of the present invention.

[0022] Figure 6 It is a structural schematic diagram of the upper shell of the utility model. DETAILED DESCRIPTION

[0023] In order to enhance the understanding of the present invention, the present invention will be described in detail below with reference to embodiments and drawings. The embodiments are only used to explain the present invention and do not limit the scope of protection of the present invention.

[0024] like Figure 1 The figure shows a protein chip for detecting respiratory viruses, which specifically includes an antigen-loaded membrane 1 on the top layer. The lower surface of the antigen-loaded membrane 1 is fixed with a reverse osmosis membrane layer 2, a water-absorbing membrane layer 3 and a leak-proof membrane layer 4 in sequence. Among them, the antigen-loaded membrane 1 is a nitrocellulose membrane, and the reverse osmosis membrane layer 2, the water-absorbing membrane layer 3 and the leak-proof membrane layer 4 are all cotton fiber layers. The difference is that the water-absorbing membrane layer 3 has a lower density and a softer material, which is conducive to water absorption due to the capillary effect; the leak-proof membrane layer 4 has a higher density and a harder material, which can absorb more liquid.

[0025] A detection area is provided on the upper surface of the antigen-loading membrane 1, comprising several antigen-loading areas 1-1 and quality control areas 1-2. Each antigen-loading area 1-1 is immobilized with a respiratory-specific antigen, specifically Chlamydia pneumoniae CP, Mycoplasma pneumoniae MP, adenovirus ADV, syncytial virus RSV, influenza B virus FLU B, and parainfluenza virus PIV. At least two antigen-loading areas 1-1 are provided for each respiratory-specific antigen, with no particular location restrictions.

[0026] Goat anti-mouse antibodies are immobilized on quality control zones 1-2, which are located at either end of antigen loading zone 1-1. The number of quality control zones at each end is different. Two quality control zones 1-2 are located at the front end of antigen loading zone 1-1, while one is located at the rear end. This facilitates positioning and identification by the biochip reader.

[0027] The specific locations of antigen loading area 1-1 and quality control area 1-2 are as follows Figure 2-4 As shown, the antigen loading area 1-1 and the quality control area 1-2 are arranged in the same row, or they can be arranged in staggered rows, as long as the distance between them is adjusted.

[0028] The protein chip is housed within a chip housing 5, which comprises an upper housing 5-1 and a lower housing 5-2. The upper housing 5-1 has a window 5-1-1, with the upper surface of the antigen-loading membrane 1 in contact with the window 5-1-1. This facilitates the dripping of the test solution into the antigen-loading membrane 1 through the window 5-1-1. A post is positioned at each of the four corners of the lower housing 5-2 to provide stable support for the chip housing 5.

[0029] During use, specific antibodies in the sample undergo a specific immune binding reaction with the antigen immobilized on the antigen-loading membrane, forming an antigen-antibody immune complex on the membrane. Colloidal gold-labeled mouse anti-human antibodies then specifically bind to this immune complex, ultimately resulting in the appearance of visible red spots. The test results are then read using a biochip reader, enabling qualitative analysis of the sample.

[0030] The embodiments disclosed in the present invention are preferred embodiments, but are not limited thereto. Ordinary technicians in this field can easily understand the spirit of the present invention based on the above embodiments and make different extensions and changes. As long as they do not deviate from the spirit of the present invention, they are all within the scope of protection of the present invention.

Claims

1. A protein chip for detecting respiratory viruses, characterized in that: The invention comprises an antigen loading membrane (1), wherein a reverse osmosis membrane layer (2), a water absorption membrane layer (3) and a leak-proof membrane layer (4) are fixed in sequence on the lower surface of the antigen loading membrane (1), and a detection area is provided on the upper surface of the antigen loading membrane (1), wherein the detection area comprises a plurality of antigen loading areas (1-1) and a quality control area (1-2), and each antigen loading area (1-1) is fixedly loaded with a respiratory tract specific antigen, wherein the respiratory tract specific antigen comprises Chlamydia pneumoniae, Mycoplasma pneumoniae, adenovirus, syncytial virus, influenza B virus and parainfluenza virus.

2. The protein chip for detecting respiratory viruses according to claim 1, characterized in that At least two antigen loading areas (1-1) are provided for each respiratory tract specific antigen.

3. The protein chip for detecting respiratory viruses according to claim 1, characterized in that The quality control area (1-2) is fixed with goat anti-mouse antibodies.

4. The protein chip for detecting respiratory viruses according to claim 3, characterized in that The quality control areas (1-2) are respectively arranged at both ends of the antigen loading area (1-1).

5. The protein chip for detecting respiratory viruses according to claim 4, characterized in that The number of quality control areas (1-2) at both ends of the antigen loading area (1-1) is different.

6. The protein chip for detecting respiratory viruses according to claim 5, characterized in that Two quality control areas (1-2) are provided at the front end of the antigen loading area (1-1), and one quality control area (1-2) is provided at the rear end.

7. The protein chip for detecting respiratory viruses according to claim 1, characterized in that The antigen loading membrane (1) is arranged in a chip housing (5), and the chip housing (5) comprises an upper housing (5-1) and a lower housing (5-2). A window (5-1-1) is arranged on the upper housing (5-1), and a column (5-2-1) is arranged on the lower housing (5-2).

8. The protein chip for detecting respiratory viruses according to claim 1, characterized in that The antigen loading membrane (1) is a nitrocellulose membrane, and the reverse osmosis membrane layer (2), the water absorption membrane layer (3) and the leak-proof membrane layer (4) are all cotton fiber membranes.

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