Biochip for detecting C-reactive protein as well as preparation method and application of biochip

Abstract

The invention discloses a biological chip for detecting C-reactive protein as well as a preparation method and application of the biological chip. The biochip comprises a solid phase carrier and an immunodetection system, the solid phase carrier comprises a substrate and a glue layer, the immunodetection system comprises a microarray and a biological material, and the biological material is used for targeted recognition of a target object to be detected and is selected from an aptamer or an antibody. The lowest detection limit value of the biochip is 1.1 ng / mL, and the biochip can be used for rapidly and ultrasensitively detecting the C-reactive protein, is suitable for rapid detection of patients by basic medical institutions, and has great significance in rapid screening of patients in remote areas or outpatient service.

Description

Technical Field

[0001] The invention belongs to the field of biological immunology and in vitro diagnosis, and particularly relates to a biochip for detecting C-reactive protein and a preparation method and application thereof. Background Art

[0002] When biological tissues are stimulated by damaging factors such as trauma and infection, and cytokines are overproduced in the human body, an inflammatory storm will occur, causing a systemic inflammatory response in a short period of time, attacking normal tissues and organs, and leading to multiple organ failure. Therefore, it is of great significance to make early identification and diagnosis of inflammatory reactions and classify and treat them as soon as possible. C-reactive protein (CRP) is a non-specific inflammatory marker and is widely used in clinical testing. The current rapid CRP test has the problem of low sensitivity, while the high-precision test has the problems of long time consumption, complicated detection process, and the need for professional testing personnel. It is necessary to develop a low-cost, ultra-sensitive rapid detection method. Summary of the invention

[0003] In order to improve the above technical problems, the present invention provides the following technical solutions:

[0004] A biochip, comprising a solid phase carrier and an immune detection system;

[0005] The solid phase carrier comprises a substrate and a glue layer, and the glue layer is arranged on the substrate;

[0006] The immunoassay system is arranged on the surface of the glue layer; the immunoassay system comprises a microarray and a biomaterial, the biomaterial is used for targeted identification of the target object to be detected, the microarray comprises an optical metamaterial, and the biomaterial is chemically modified on the surface of the optical metamaterial;

[0007] The biological material is selected from aptamers or antibodies.

[0008] According to an embodiment of the present invention, the substrate is selected from plastic substrates, for example, selected from PET, PVC, PP, PE, PS, preferably PET.

[0009] According to an embodiment of the present invention, the glue layer is prepared from water-soluble glue.

[0010] According to an embodiment of the present invention, the water-soluble glue is selected from at least one of polyvinyl alcohol-based water-based adhesives, vinyl acetate-based water-based adhesives, acrylic water-based adhesives, polyurethane water-based adhesives, epoxy water-based adhesives, phenolic water-based adhesives, and the like.

[0011] According to an embodiment of the present invention, the microarray comprises a dot matrix formed by self-assembly of optical metamaterials.

[0012] According to an embodiment of the present invention, the optical metamaterial is formed of periodically arranged microspheres of a photonic crystal structure.

[0013] According to an embodiment of the present invention, the photonic crystal structure microspheres are selected from at least one of polystyrene, poly(methyl methacrylate-acrylic acid-styrene), and silicon dioxide.

[0014] According to an embodiment of the present invention, the particle size of the photonic crystal structure microsphere is 290 nm.

[0015] According to an embodiment of the present invention, the dot matrix of the microarray is selected from a 9-dot matrix, a 16-dot matrix, and a 28-dot matrix.

[0016] According to an embodiment of the present invention, the photonic crystal structure microspheres are arranged periodically and neatly on the substrate.

[0017] According to an embodiment of the present invention, the aptamer is selected from amino-modified aptamers, preferably amino-modified aptamers that can capture C-reactive protein, and the antibody is selected from antibodies that can capture C-reactive protein.

[0018] According to an embodiment of the present invention, the antibody is selected from a fluorescently labeled antibody or a non-fluorescently labeled antibody, and is preferably an antibody that can capture C-reactive protein.

[0019] According to an embodiment of the present invention, the target object to be detected is selected from a sample containing C-reactive protein, for example, selected from a human serum sample.

[0020] The present invention also provides a method for preparing the above biochip, the specific steps of which are as follows:

[0021] (1) providing a glue layer on a substrate, further forming a patterned microarray on the glue layer by self-assembly of an optical metamaterial, and then sintering and curing the microarray;

[0022] (2) in situ printing an activator on the surface of the microarray after sintering and curing in step (1), activating the microarray, connecting the biomaterial to the microarray, and then blocking the remaining connection sites with a blocking agent to obtain the biochip.

[0023] According to an embodiment of the present invention, the preparation method of the microarray comprises: printing ink containing optical metamaterials on the glue layer to form a lattice including the photonic crystal structure microspheres to obtain the microarray.

[0024] According to an embodiment of the present invention, the optical metamaterial has the meaning as described above, for example, at least one selected from polystyrene, poly(methyl methacrylate-acrylic acid-styrene), and silicon dioxide.

[0025] According to an embodiment of the present invention, the water-soluble glue layer is disposed between the substrate and the optical metamaterial.

[0026] According to an embodiment of the present invention, the water-soluble glue layer is obtained by uniformly coating water-soluble glue on the substrate.

[0027] According to an embodiment of the present invention, the water-soluble glue is selected from at least one of polyvinyl alcohol-based water-based adhesives, vinyl acetate-based water-based adhesives, acrylic water-based adhesives, polyurethane water-based adhesives, epoxy water-based adhesives, phenolic water-based adhesives, and the like.

[0028] According to an embodiment of the present invention, in step (1), the curing and sintering temperature is 100 to 120° C., preferably 110° C.; the curing and sintering time is 10 to 30 minutes, preferably 12 to 20 minutes.

[0029] According to an embodiment of the present invention, in step (2), the concentration of the activator is 1 to 60 mg / mL, for example, 5 to 30 mg / mL.

[0030] According to an embodiment of the present invention, in step (2), the activator is 1-ethyl-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysulfosuccinimide (NHS), and the mass concentration ratio thereof is 5-20:1-10, for example, 10:3 or 4:1, preferably 10:3. Preferably, the preparation method of the activator comprises: before in-situ printing, EDC and NHS are mixed uniformly in a buffer solution system in a mass ratio of 5-20:1-10 (for example, 10:3 or 4:1) and then used.

[0031] According to an embodiment of the present invention, in step (2), the activation time is 10 min to 60 min, preferably 20 min.

[0032] According to an embodiment of the present invention, in step (2), the biological material is selected from antibodies or amino-modified aptamers.

[0033] According to an embodiment of the present invention, the concentration of the amino-modified aptamer is 0.5-2 μmol / L, preferably 1 μmol / L, and the concentration of the antibody is 10-100 μg / mL, preferably 50 μg / mL.

[0034] According to an embodiment of the present invention, in step (2), the biomaterial is connected to the microarray via an amidation reaction. Preferably, the conditions of the amidation reaction are: temperature of 2-8°C; time of 10-20h.

[0035] According to an embodiment of the present invention, in step (2), the blocking agent is bovine serum albumin (BSA) and / or sodium caseinate.

[0036] According to an embodiment of the present invention, in the blocking agent, the concentration of bovine serum albumin is 5 to 20 mg / mL, preferably 10 mg / mL.

[0037] According to an embodiment of the present invention, in the blocking agent, the concentration of sodium caseinate is 0.01-1 mg / mL, preferably 0.05 mg / mL.

[0038] The present invention also provides the use of the above biochip, preferably for detecting target analytes, such as C-reactive protein.

[0039] The present invention also provides a detection method for the above-mentioned biochip, which comprises: mixing the detection sample with the detection probe, dripping the mixture onto the biochip, incubating the mixture, and detecting the fluorescence intensity of the biochip.

[0040] According to an embodiment of the present invention, the incubation is carried out at room temperature (10-30° C., such as 20° C.); the incubation time is 5 to 40 min, preferably 10 min.

[0041] According to an embodiment of the present invention, the detection refers to testing the fluorescence signal of the biochip under a portable fluorescence tester.

[0042] According to an embodiment of the present invention, the flushing liquid is deionized water, and the flushing times are 3 to 5 times.

[0043] According to an embodiment of the present invention, the test sample is selected from a sample containing a target analyte, for example, a sample containing C-reactive protein.

[0044] According to an embodiment of the present invention, the test sample is, for example, a serum sample.

[0045] According to an embodiment of the present invention, the detection probe is selected from a detection aptamer labeled with a fluorescent dye or a detection antibody labeled with a fluorescent dye.

[0046] According to an embodiment of the present invention, the emission wavelength of the fluorescent dye matches the particle size of the microspheres of the photonic crystal structure.

[0047] For example, the fluorescent dye is a non-sulfonated cyanine dye, such as Cy3, Cy5 or Cy7.

[0048] Exemplarily, the detection probe is selected from a Cy5-labeled aptamer or a Cy5-labeled antibody solution, and its concentration is, for example, 1 μmol / L or 50 μg / mL.

[0049] The present invention also provides a portable detection device, which includes the above-mentioned biochip.

[0050] According to an embodiment of the present invention, the portable detection device is a kit, and the kit includes the above-mentioned biochip.

[0051] According to an embodiment of the present invention, the kit further comprises a diluent, a washing solution, a detection probe, a pipette and a storage medium.

[0052] The present invention also provides uses of the above-mentioned biochip and / or portable detection device, for example, for detecting C-reactive protein.

[0053] Beneficial Effects

[0054] 1. The present invention provides a biochip and detection platform that can detect C-reactive protein quickly, easily and with high sensitivity. The biochip of the present invention can be used in portable detection equipment, which is cheap and portable, and can be used to complete the C-reactive protein detection of patients in a timely manner without the use of complex equipment or professional data analysis.

[0055] 2. The minimum detection limit of the biochip of the present invention is 1.1 ng / mL, and its detection sensitivity is higher than that of the colloidal gold method, the detection time is shorter than that of the enzyme-linked immunosorbent assay (ELISA) method (whose detection time is usually more than 90 minutes), and the cost is lower than that of the colloidal gold method.

[0056] 3. The biochip of the present invention is suitable for rapid testing of patients in primary medical institutions, and is of great significance for rapid screening of patients in remote areas or outpatient clinics. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] Figure 1 Schematic diagram of the preparation of a biochip for rapid and ultra-sensitive detection of C-reactive protein; Figure numerals: 1-plastic substrate, 2-water-soluble glue layer, 3-optical metamaterial microarray.

[0058] Figure 2 The standard curve of fluorescence intensity of C-reactive protein biochip. DETAILED DESCRIPTION

[0059] The technical scheme of the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the following embodiments are only exemplary descriptions and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are included in the scope that the present invention is intended to protect.

[0060] Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

[0061] Example 1

[0062] Preparation of C-reactive protein biochip:

[0063] When a double antibody sandwich detection system is used to detect C-reactive protein, a capture aptamer modified with an amino group at the 5' end and a detection aptamer modified with Cy5 at the 3' end are selected, the sequences of which are shown in Table 1 and can capture C-reactive protein. Therefore, in this embodiment, an optical metamaterial matching the emission wavelength of Cy5 is selected to print an optical metamaterial microarray with a periodic arrangement, and the optical metamaterial is selected from polystyrene microspheres with a particle size of 290 nm and surface modified with carboxyl groups.

[0064] The specific preparation method is as follows:

[0065] (1) The optical metamaterial ink used for printing is an ink containing monodisperse poly(methyl methacrylate-acrylic acid-styrene) latex spheres (mass concentration 10%), ethylene glycol (volume fraction 10%) and surfactant BYK-3400 (mass concentration 1‰). The optical metamaterial ink is printed onto a PET substrate coated with water-soluble glue using a Nordson PICO Pulse XP&PROPlus4 dispenser. After drying, it is heated and sintered at 110°C for 15 minutes to obtain an optical metamaterial microarray.

[0066] (2) In a PBS buffer solution system (pH 7.2-7.4), a mixed solution of EDC / NHS (concentrations of 10 mg / mL and 3 mg / mL, respectively) was in situ inkjet printed onto the surface of the optical metamaterial array at room temperature. 30 μL of the mixed solution was printed at each point and activated at room temperature for 20 min. Then, the 5'-terminal amino-modified capture aptamer (1 μmol / L) was incubated at room temperature for 2 h.

[0067] (3) After the incubation is completed, the chip is washed 3-5 times with deionized water to wash away excess uncoupled capture aptamers, thereby obtaining a C-reactive protein detection biochip.

[0068] (4) Use a blocking agent (1% BSA solution) solution to block the above biochip, completely immerse the aptamer chip cleaned in step (3) in the blocking agent, and block at room temperature for 30 minutes to block the remaining binding sites to reduce nonspecific adsorption. After blocking, wash the chip with deionized water 3-5 times to wash away the excess blocking agent solution. Place the blocked biochip in a sealed culture dish and store it in a 4°C refrigerator overnight.

[0069] Table 1

[0070]

[0071] Example 2

[0072] The method for using the C-reactive protein biochip of Example 1 to detect recombinant C-reactive protein antigen is as follows:

[0073] (1) Recombinant C-reactive protein antigen (stock concentration of 200 μg / mL, purchased from Xiamen Tongrenxin Biotechnology Co., Ltd.) was diluted 40 times, 200 times, 400 times, 2000 times, 4000 times, 20000 times, and 40000 times with PBS buffer to obtain a series of recombinant C-reactive protein antigen solutions (concentrations of 5 μg / mL, 1 μg / mL, 500 ng / mL, 100 ng / mL, 50 ng / mL, 10 ng / mL, and 5 ng / mL, respectively) and mixed with a 3'-end Cy5-modified detection aptamer (1 μmol / L) to obtain standard antigen samples of recombinant C-reactive protein with different concentrations.

[0074] (2) Take standard antigen samples of recombinant C-reactive protein of different concentrations prepared in step (1), and drip them onto the C-reactive protein biochip prepared in Example 1, and incubate at room temperature for 10 minutes.

[0075] (3) After the reaction is completed, the biochip is washed 3 to 5 times with deionized water to wash away the unrecognized C-reactive protein standard antigen sample on the biochip.

[0076] (4) Use a portable fluorescence detector to test the fluorescence data of the biochip after cleaning in step (3). The test results are shown in Table 2, and the test results are fitted to obtain a standard curve of the fluorescence intensity of the C-reactive protein biochip detection, such as Figure 2 shown.

[0077] Table 2

[0078]

[0079] According to the fluorescence intensity values ​​obtained by the biochip detection of C-reactive protein in Table 2, it can be seen that the detection limit of the biochip of the present invention is 5 ng / mL.

[0080] Example 3

[0081] Another method for preparing a biochip for detecting C-reactive protein is as follows:

[0082] A double antibody sandwich detection system is used to detect C-reactive protein. The double antibody includes a capture antibody that can capture C-reactive protein and a detection antibody labeled with Cy5, and polystyrene with a particle size of 290 nm is selected as the photonic crystal microspheres that match the emission wavelength of Cy5.

[0083] The specific preparation process is as follows:

[0084] (1) The optical metamaterial ink was printed onto a PET substrate coated with water-soluble glue using a Nordson PICO Pulse XP&PROPlus4 dispensing machine. After drying, the ink was heated and sintered at 110°C for 15 minutes to obtain an optical metamaterial microarray.

[0085] (2) EDC / NHS was mixed in a pH=4 MES buffer at a mass ratio of 1:1 to obtain a mixed solution, and the concentrations of EDC and NHS were 40 mg / mL and 40 mg / mL, respectively; 20 μL of the mixed solution was printed at each point on the photonic crystal array in step (1) at room temperature, and activated at room temperature for 10 min. The chip was rinsed with deionized water 3 times.

[0086] (3) The capture antibody (50 μg / mL) was incubated on the photonic crystal microarray after cleaning in (2) at room temperature for 2 h.

[0087] (4) After the incubation is completed, the chip is washed three times with deionized water to wash away excess uncoupled antibodies, thereby obtaining an antibody chip for detecting C-reactive protein.

[0088] (5) The above antibody chip was blocked: the antibody chip was completely immersed in a blocking agent (1% BSA solution and 0.005% sodium casein solution) at room temperature for 0.5 h, so that the protein in the blocking agent occupied the uncoupled area of ​​the antibody chip, reducing the direct binding of the labeled antibody to the chip surface and reducing nonspecific adsorption; then the antibody chip was washed three times with deionized water to wash away the excess blocking agent. The blocked antibody chip was placed in a sealed culture dish and stored in a 4°C refrigerator overnight.

[0089] Example 4

[0090] The method for testing patient serum is as follows:

[0091] (1) Patient sera with known different CRP concentrations were diluted 200 times with PBS buffer to obtain a series of serum dilutions with different CRP concentrations (1.1 ng / mL, 8.9 ng / mL, 29.6 ng / mL, 68.7 ng / mL, 285 ng / mL, 399 ng / mL), which were mixed with Cy5-labeled detection antibody (50 μg / mL).

[0092] (2) 10 μL of CRP serum dilutions of different concentrations prepared in step (1) were added dropwise to the detection chip prepared in Example 3 and incubated for 10 min at room temperature.

[0093] (3) After the reaction is completed, the detection chip is cleaned with deionized water for 3 times to wash away the excess mixed solution on the detection chip.

[0094] (4) After the chip is tested, the fluorescence data is tested using a portable instrument.

[0095] Table 3 shows the fluorescence intensity values ​​of different concentrations of C-reactive protein detected by the C-reactive protein biochip, indicating that the detection limit of the chip is 1.1 ng / mL.

[0096] Table 3

[0097]

[0098] Example 5

[0099] C-reactive protein biochip preparation biochip kit and method for detecting C-reactive protein are as follows:

[0100] 1. Biochip kit

[0101] The biochip kit for rapid ultrasensitive detection of C-reactive protein in this embodiment includes a biochip, a diluent, a flushing solution, a detection probe, a pipette and a storage medium; wherein the biochip is the C-reactive protein biochip prepared in Example 1.

[0102] 2. How to use the kit:

[0103] (1) The target substance to be detected (human serum sample) was mixed with the detection probe (Cy5-labeled aptamer solution, 1 μmol / L), added dropwise to the C-reactive protein aptamer chip, and incubated at room temperature for 10 min.

[0104] (2) The C-reactive protein aptamer chip is washed with a flushing solution for 3 to 5 times to wash away the excess unidentified target detection substances on the C-reactive protein biochip.

[0105] (3) The C-reactive protein aptamer chip identified in step (2) is placed under a portable fluorescence detector, and the excitation light required for the specific labeled fluorescence is selected for excitation, and data is read, and the read value is compared with the fluorescence intensity standard curve in Example 2.

[0106] 3. Storage of the kit

[0107] The kit should be stored at 4°C.

[0108] Comparative Example 1

[0109] 1. The preparation method of the C-reactive protein biochip of this comparative example is basically the same as that of Example 1, except that:

[0110] In step (2), different activators were used: EDC (4 mg / mL) solution was prepared in PBS buffer solution system (pH 7.2-7.4), NHS (1 mg / mL) solution was prepared in anhydrous ethanol, the two solutions were mixed in equal volumes, and the mixed solution was in situ inkjet printed on the surface of the optical metamaterial array at room temperature, 30 μL per drop, and activated at room temperature for 20 minutes. Then, the amino-modified capture aptamer (1 μmol / L) was incubated at room temperature for 2 hours.

[0111] 2. Detection limit of biochip:

[0112] Referring to the detection method of Example 2, the detection limit of the C-reactive protein biochip of this comparative example was tested, except that: the recombinant C-reactive protein antigen series solutions prepared in Example 2 were taken, with concentrations of 10 ng / mL, 100 ng / mL, and 50 μg / mL, and were respectively mixed with Cy5-labeled detection aptamer (1 μmol / L) to obtain standard antigen samples of recombinant C-reactive protein of different concentrations, and the samples were dropped onto the C-reactive protein biochip of this comparative example; the remaining steps were the same as in Example 2, and the test results are shown in Table 4.

[0113] Biochip Biochip

[0114] Table 4

[0115]

[0116] According to the fluorescence intensity values ​​in Table 4, the detection limit of the biochip prepared in this comparative example is 10 ng / mL.

[0117] The above is a description of the exemplary embodiments of the present invention. However, the protection scope of the present application is not limited to the above embodiments. Any modification, equivalent substitution, improvement, etc. made by those skilled in the art within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A biochip, It is characterized in that The biochip includes a solid phase carrier and an immune detection system; The solid phase carrier comprises a substrate and a glue layer, and the glue layer is arranged on the substrate; The immunoassay system is arranged on the surface of the glue layer; the immunoassay system comprises a microarray and a biomaterial, the biomaterial is used for targeted identification of the target object to be detected, the microarray comprises an optical metamaterial, and the biomaterial is chemically modified on the surface of the optical metamaterial; The biological material is selected from aptamers or antibodies.

2. The biochip according to claim 1, It is characterized in that The substrate is selected from plastic substrates. Preferably, the glue layer is prepared from water-soluble glue. Preferably, the water-soluble glue is selected from at least one of polyvinyl alcohol-based water-based adhesives, vinyl acetate-based water-based adhesives, acrylic acid-based water-based adhesives, polyurethane-based water-based adhesives, epoxy water-based adhesives, phenolic water-based adhesives, and the like. Preferably, the microarray comprises a dot matrix formed by self-assembly of optical metamaterials. Preferably, the optical metamaterial is formed by periodically arranged microspheres of a photonic crystal structure. Preferably, the photonic crystal structure microspheres are selected from at least one of polystyrene, poly(methyl methacrylate-acrylic acid-styrene), and silicon dioxide. Preferably, the photonic crystal structure microspheres are arranged periodically and neatly on the substrate.

3. The biochip according to claim 1 or 2, It is characterized in that The aptamer is selected from amino-modified aptamers. Preferably, the antibody is selected from a fluorescently labeled antibody or a non-fluorescently labeled antibody. Preferably, the target substance to be detected is selected from a sample containing C-reactive protein.

4. The method for preparing the biochip according to any one of claims 1 to 3, It is characterized in that The specific steps of the preparation method are as follows: (1) providing a glue layer on a substrate, further forming a patterned microarray on the glue layer by self-assembly of an optical metamaterial, and then sintering and curing the microarray; (2) in situ printing an activator on the surface of the microarray after sintering and curing in step (1), activating the microarray, connecting the biomaterial to the microarray, and then blocking the remaining connection sites with a blocking agent to obtain the biochip.

5. The preparation method according to claim 4, It is characterized in that The preparation method of the microarray comprises: printing ink containing optical metamaterial on the glue layer to form a dot matrix including the photonic crystal structure microspheres to obtain the microarray. Preferably, the water-soluble glue layer is disposed between the substrate and the optical metamaterial. Preferably, the water-soluble glue layer is obtained by uniformly coating water-soluble glue on the substrate. Preferably, the water-soluble glue is selected from at least one of polyvinyl alcohol-based water-based adhesives, vinyl acetate-based water-based adhesives, acrylic acid-based water-based adhesives, polyurethane-based water-based adhesives, epoxy water-based adhesives, phenolic water-based adhesives, and the like.

6. The preparation method according to claim 4 or 5, It is characterized in that In step (1), the curing and sintering temperature is 100 to 120° C.; and the curing and sintering time is 10 to 30 minutes. Preferably, in step (2), the concentration of the activator is 1 to 60 mg / mL. Preferably, in step (2), the activator is 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxysulfosuccinimide, and the mass concentration ratio thereof is 5-20:1-10. Preferably, in step (2), the activation time is 10 min to 60 min. Preferably, in step (2), the biological material is selected from antibodies or amino-modified aptamers. Preferably, the concentration of the amino-modified aptamer is 0.5-2 μmol / L, and the concentration of the antibody is 10-100 μg / mL. Preferably, in step (2), the biological material is connected to the microarray via an amidation reaction. Preferably, in step (2), the blocking agent is bovine serum albumin (BSA) and / or sodium caseinate. Preferably, in the blocking agent, the concentration of bovine serum albumin is 5 to 20 mg / mL. Preferably, in the blocking agent, the concentration of sodium caseinate is 0.01-1 mg / mL.

7. Use of the biochip according to any one of claims 1 to 3 in detecting a target analyte.

8. The biochip detection method according to any one of claims 1 to 3, It is characterized in that The detection method comprises: mixing the detection sample with the detection probe, dripping the mixture onto the biochip, incubating the mixture, and detecting the fluorescence intensity of the biochip.

9. A portable detection device, It is characterized in that The detection device comprises the biochip according to any one of claims 1-3.

10. Use of the biochip and / or portable detection device according to any one of claims 1 to 3 in the detection of C-reactive protein.

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