CN111999507B - Fluorescent immunochromatography test paper for detecting novel coronavirus antibody - Google Patents
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/585—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with a particulate label, e.g. coloured latex
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- G01N2469/20—Detection of antibodies in sample from host which are directed against antigens from microorganisms
Abstract
The invention discloses a fluorescent immunochromatographic test paper for detecting novel coronavirus antibodies. The combining pad of the test paper is coated with the anti-human IgM antibody marked by the time-resolved fluorescence microsphere; the preparation method of the bonding pad comprises the following steps: mixing time-resolved fluorescent microsphere with anti-human IgM antibody, adding boric acid buffer solution containing 0.1-0.5% BSA for blocking; centrifuging and collecting precipitate; re-suspending the precipitate with boric acid buffer solution containing 0.1-0.5% BSA to obtain labeling solution; spraying the marking solution on the film, and drying to obtain a bonding pad; the boric acid buffer solution is pH7.0-8.5, 10-100mM boric acid buffer solution. Experiments prove that the fluorescent immunochromatography test paper provided by the invention can be used for detecting whether a sample contains a novel coronavirus antibody, and has the advantages of simplicity in operation, short time consumption, high accuracy, good specificity and high sensitivity. The fluorescent immunochromatographic test paper provided by the invention has important application value in rapidly detecting novel coronavirus antibodies on site.
Description
Technical Field
The invention belongs to the field of biotechnology, and particularly relates to fluorescent immunochromatography test paper for detecting novel coronavirus antibodies.
Background
Coronaviruses are a class of enveloped single-stranded positive-stranded RNA viruses, and are known by the corona-like morphology of the virions under electron microscopy. The host range of coronaviruses mainly includes birds and mammals, and seven kinds of coronaviruses that can infect humans have been identified so far, among which the emergence of SARS virus (SARS-CoV), middle east respiratory syndrome coronavirus (MERS-CoV) and novel coronavirus (SARS-CoV-2) pose a great threat to world economic health. The novel coronavirus belongs to coronaviridae and coronaviridae, can cause symptoms of different degrees after infecting people, and has the main manifestations of fever, hypodynamia and dry cough, and severe cases of pneumonia, acute respiratory distress syndrome and even death. The main modes of transmission of the novel coronaviruses are droplet transmission and contact transmission.
In the detection of novel coronaviruses, it is initially achieved by detecting whether or not a sample contains viral nucleic acids, but the positive rate is reduced due to sample collection, reagents, operations, and the like. The suspected cases with pneumonia imaging characteristics are used as clinical diagnosis case standard. Similar histological pathology may occur for different types of pneumonia, and CT detection is at risk of cross-infection.
Disclosure of Invention
The object of the present invention is to detect novel coronaviruses.
The invention first protects a fluorescent immunochromatographic test paper for detecting novel coronavirus antibodies, and the binding pad of the fluorescent immunochromatographic test paper is coated with anti-human IgM antibodies marked by time-resolved fluorescent microspheres;
the preparation method of the bonding pad can be as follows:
(1) Mixing time-resolved fluorescent microsphere with anti-human IgM antibody, adding boric acid buffer solution containing 0.1-0.5% (such as 0.1-0.3%, 0.3-0.5%, 0.1%, 0.3% or 0.5%) BSA, and sealing;
(2) After the step (1) is completed, centrifuging and collecting sediment;
(3) After step (2) is completed, the precipitate is resuspended with boric acid buffer containing 0.1-0.5% (e.g., 0.1-0.3%, 0.3-0.5%, 0.1%, 0.3%, or 0.5%) BSA to obtain a labeling solution;
(4) Spraying the marking solution on the membrane, and drying to obtain a bonding pad coated with the anti-human IgM antibody marked by the time-resolved fluorescence microsphere;
The boric acid buffer is pH7.0-8.5 (such as pH7.0-7.5, pH7.5-8.0, pH8.0-8.5, pH7.0, pH7.5, pH8.0 or pH 8.5), 10-100mM (such as 10-25mM, 25-50mM, 50-100mM, 10mM, 25mM, 50mM or 100 mM) boric acid buffer.
In the above-mentioned fluorescence immunochromatography test paper, the spraying of the labeling solution on the membrane may be spraying of the labeling solution on a glass cellulose membrane.
In the above-mentioned fluorescence immunochromatographic test strip, the boric acid buffer solution may specifically be a boric acid buffer solution of pH8.0 and 25 mM.
In the above fluorescent immunochromatographic test paper, the time-resolved fluorogenic microsphere may be a europium-containing time-resolved fluorogenic microsphere.
In the fluorescent immunochromatographic test paper, the antibody of the anti-human IgM antibody can be coated at the position of the quality control line on the reaction membrane. The detection line positions may be coated with novel coronavirus antigens.
Any of the novel coronavirus antigens described above may be novel coronavirus NP antigens.
Any of the above anti-human IgM antibodies may be a sheep anti-human IgM antibody, a mouse anti-human IgM antibody or a rabbit anti-human IgM antibody.
The antibody of any of the above-mentioned goat anti-human IgM antibodies may specifically be a rabbit anti-goat IgM antibody.
Any of the above-described reactive films may be nitrocellulose films.
The preparation method of the sheep anti-human IgM antibody can be referred to as the following documents: preparation of goat anti-human mu-chain serum, chinese public health, 1985,4 (4): 38-40.
The preparation method of the rabbit anti-sheep IgM antibody comprises the following steps of: a simple method for preparing goat anti-rabbit IgG antibodies, university of capital medical science, 1998, 19 (1).
Any one of the above fluorescence immunochromatography test papers can specifically comprise a sample pad (made of glass cellulose membrane), a combination pad, a nitrocellulose membrane coating a quality control line and a detection line, and a water absorption pad (such as filter paper), and the specific structure is shown in fig. 1.
In any one of the above-mentioned fluorescence immunochromatographic test papers, the preparation method of the binding pad may specifically be as follows:
(1-1) adding 800 mu L of goat anti-human IgM antibody with the concentration of 1mg/ml into 6-10ml of solution containing europium time-resolved fluorescence microsphere, and uniformly mixing for 60min on a rotary uniformly mixer;
(1-2) adding boric acid buffer containing 0.3% BSA, blocking overnight;
(1-3) centrifuging at 15000rpm for 40min, and collecting the precipitate;
(1-4) re-suspending and precipitating with 12mL of 0.1% (m/v) BSA boric acid buffer solution to obtain a europium-containing time-resolved fluorescence microsphere-labeled goat anti-human IgM antibody solution;
(1-5) uniformly spraying a europium-containing time-resolved fluorescence microsphere marked sheep anti-human IgM antibody solution on a glass fiber membrane, and then freeze-drying by a freeze dryer to obtain a bonding pad;
The boric acid buffer may specifically be a pH8.0, 25mM boric acid buffer.
In any one of the above-mentioned fluorescence immunochromatography test papers, the preparation method of the nitrocellulose membrane coating the quality control line and the detection line specifically comprises the following steps:
(2-1) diluting the novel coronavirus NP antigen with 0.01M PBS buffer (pH 7.4) to obtain a detection line solution having a concentration of 3mg/mL for coating the detection line;
(2-2) diluting rabbit anti-sheep IgM antibody with 0.01M PBS buffer (pH 7.4) to obtain a quality control line solution with the concentration of 2mg/mL, and coating the quality control line;
and (2-3) taking a quality control line solution and a detection line solution, respectively and uniformly spraying the quality control line solution and the detection line solution on a nitrocellulose membrane (the spraying amount is 10 mu L/cm 2), forming a quality control line (C) and a detection line (T) which are mutually separated, and drying at 37 ℃ for 3-5h to obtain the nitrocellulose membrane coated with the quality control line and the detection line. And the interval between the quality control line and the detection line is 0.5cm on the nitrocellulose membrane coated with the quality control line and the detection line.
The invention also provides a method for detecting whether the liquid to be detected contains the novel coronavirus antibody by adopting the fluorescent immunochromatographic test paper.
The method for detecting whether the liquid to be detected contains the novel coronavirus antibody by adopting the fluorescence immunochromatography test paper disclosed by the invention specifically comprises the following steps: immersing the sample pad of the fluorescent immunochromatographic test paper in the liquid to be tested, standing for more than 10min (such as 10min and 15 min), irradiating the reaction membrane with fluorescence (specifically, fluorescence with excitation wavelength of 365 nm), observing red reaction lines appearing on the reaction membrane, and judging as follows:
if the detection line and the quality control line show red reaction lines, the liquid to be detected contains novel coronavirus antibodies;
If the red reaction line is displayed at the quality control line and the red reaction line is not displayed at the detection line, the liquid to be detected does not contain the novel coronavirus antibody;
if the red reaction line is not displayed at the quality control line, the result is invalid.
The method for detecting whether the liquid to be detected contains the novel coronavirus antibody by adopting the fluorescence immunochromatography test paper disclosed by the invention specifically comprises the following steps: immersing the sample pad of the fluorescent immunochromatographic test paper into the liquid to be tested, standing for more than 10min (such as 10min and 15 min), detecting fluorescent values at the detection line and the quality control line, and judging as follows:
If the fluorescence values of the detection line and the quality control line are both greater than 50, the liquid to be detected contains a novel coronavirus antibody;
If the fluorescence value at the quality control line is greater than 50 and the fluorescence value at the detection line is less than 50, the liquid to be detected does not contain the novel coronavirus antibody;
If the fluorescence value at the quality control line is 50 or less, the result is invalid.
In the second method, the fluorescence values of the detection line and the quality control line may be fluorescence values of the detection line and the quality control line detected by a dry fluorescence detector.
In any of the above methods, the test solution may be serum or a serum dilution.
Any of the methods described above are useful for diagnosis and treatment of non-disease.
The method of any one of the above, wherein the presence of the novel coronavirus antibody can mean infection or infection with the novel coronavirus.
Any one of the above fluorescence immunochromatography test papers is used for diagnosis and treatment of non-diseases.
Experiments prove that the fluorescent immunochromatography test paper provided by the invention can be used for detecting whether a sample contains a novel coronavirus antibody, and has the advantages of simplicity in operation, short time consumption, high accuracy, good specificity and high sensitivity. The fluorescent immunochromatographic test paper provided by the invention has important application value in rapidly detecting whether the serum contains a novel coronavirus antibody or not on site.
Drawings
FIG. 1 is a schematic structural diagram of a fluorescent immunochromatographic test strip for detecting novel coronavirus antibodies.
Detailed Description
The following examples facilitate a better understanding of the present invention, but are not intended to limit the same.
The experimental methods in the following examples are conventional methods unless otherwise specified.
The test materials used in the examples described below, unless otherwise specified, were purchased from conventional Biochemical reagent companies.
The quantitative tests in the following examples were all set up in triplicate and the results averaged.
Example 1 preparation of fluorescent immunochromatographic test paper for detecting novel coronavirus antibody
1. Preparation of bond pads
1. BSA was dissolved in boric acid buffer to obtain working solution 1 having a concentration of 0.3% (m/v) BSA and working solution 2 having a concentration of 0.1% (m/v) BSA.
2. To 6-10ml of a solution of europium-containing time-resolved fluorescent microsphere (product of Thermo Scientific TM Co., ltd., product No. 93470520010150; particle size 200 nm), 800. Mu.L of goat anti-human IgM antibody (preparation method of goat anti-human IgM antibody is referred to the following literature: preparation of goat anti-human. Mu. Chain serum, china public health, 1985,4 (4): 38-40) was added, and the mixture was homogenized by a rotary kneader for 60 minutes.
3. After the completion of step 2, working solution 1 was added and blocked overnight.
4. After completion of step 3, the mixture was centrifuged at 15000rpm for 40min, and the precipitate was collected.
5. After the step 4 is completed, 12mL of working solution 2 is used for resuspension and precipitation, and the solution of the sheep anti-human IgM antibody marked by the europium-containing time-resolved fluorescent microsphere is obtained.
6. Uniformly spraying a solution of the sheep anti-human IgM antibody marked by europium-containing time-resolved fluorescent microspheres on a glass cellulose membrane, and then freeze-drying by a freeze dryer to obtain the bonding pad.
2. Preparation of nitrocellulose membrane coated with quality control line and detection line
1. The novel coronavirus NP antigen (product of the eastern ocean medical institute, cat# B35-Ag 1) was diluted with 0.01M PBS buffer (pH 7.4) to give a detection line solution with a concentration of 3mg/mL for coating the detection line.
2. The rabbit anti-goat IgM antibody was diluted with 0.01M PBS buffer (pH 7.4) (preparation method of rabbit anti-goat IgM antibody referring to 1 simple method for preparing goat anti-goat IgG antibody, university of first medical science, report 1998, 19 (1)), to obtain a 2mg/mL solution of the quality control line for coating the quality control line.
3. And (3) taking a quality control line solution and a detection line solution, respectively and uniformly spraying the quality control line solution and the detection line solution on a nitrocellulose membrane (the spraying amount is 10 mu L/cm 2), so as to form a quality control line (C) and a detection line (T) which are mutually separated, and drying at 37 ℃ for 3-5h, thus obtaining the nitrocellulose membrane coated with the quality control line and the detection line. And the interval between the quality control line and the detection line is 0.5cm on the nitrocellulose membrane coated with the quality control line and the detection line.
3. Assembly of fluorescent immunochromatographic test paper for detecting novel coronavirus antibody
The fluorescent immunochromatographic test paper for detecting the novel coronavirus antibody consists of a sample pad (made of a glass cellulose membrane), a binding pad, a nitrocellulose membrane coating a quality control line and a detection line and a water absorption pad (such as filter paper).
The assembling steps of the fluorescent immunochromatographic test paper for detecting the novel coronavirus antibody are as follows: the method comprises the steps of adhering a water absorption pad to one end (the overlapping length is 0.2 cm) of a nitrocellulose membrane coated with a quality control line and a detection line by using double-sided adhesive, adhering one end (the overlapping length is 0.2 cm) of a bonding pad to the other end of the nitrocellulose membrane coated with the quality control line and the detection line by using double-sided adhesive, adhering a sample pad (the overlapping length is 0.2 cm) to the other end of the bonding pad by using double-sided adhesive, adhering the sample pad to a plastic backboard (such as a PVC (polyvinyl chloride) substrate) by using double-sided adhesive, and cutting the sample pad into fluorescent immunochromatographic test paper with the width of 3mm for detecting novel coronavirus antibodies. Sealing the fluorescence immunochromatography test paper for detecting the novel coronavirus antibody by using a plastic sealing bag, and then placing the sealed paper in a drying box for storage for later use.
The structural schematic diagram of the fluorescent immunochromatographic test paper for detecting the novel coronavirus antibody is shown in figure 1. Sample addition holes may also be provided in the sample pad as desired.
4. Using method and judgment standard of fluorescent immunochromatography test paper for detecting novel coronavirus antibody
Taking fluorescent immunochromatography test paper for detecting novel coronavirus antibodies, adding 100 mu L of to-be-detected liquid (to-be-detected serum or to-be-detected serum diluent) into a sample pad (or a sample adding hole), sucking the to-be-detected liquid by the sample pad, continuing to move to the upper end, enabling the to-be-detected liquid to act with sheep anti-human IgM antibodies marked by europium-containing time-resolved fluorescent microspheres when flowing through a binding pad, and then performing migration to a nitrocellulose membrane coated with a quality control line and a detection line for 15min. The nitrocellulose membrane was irradiated with a fluorescent torch having an excitation wavelength of 365 nm. The following judgment is carried out according to the condition of the red reaction lines displayed on the film:
1. If red reaction lines appear at the detection line and the quality control line on the nitrocellulose membrane, the liquid to be tested contains a novel coronavirus antibody, namely, the person to be tested is infected with the novel coronavirus; the principle is that a novel coronavirus antibody in a liquid to be detected is combined with a sheep anti-human IgM antibody marked by europium-containing time-resolved fluorescence microspheres on a combination pad to form an immune complex, the complex flows through a detection line to be combined with a novel coronavirus NP antigen, a bridging europium-containing time-resolved fluorescence microsphere, so that red reaction lines are displayed at the detection line, and the complex flows through a quality control line to be combined with a rabbit anti-sheep IgM antibody, and a bridging europium-containing time-resolved fluorescence microsphere, so that red reaction lines are displayed at the quality control line;
2. If only a red reaction line appears at the quality control line on the nitrocellulose membrane, the liquid to be tested does not contain a novel coronavirus antibody, i.e. the person to be tested is not infected with the novel coronavirus; the principle is that the liquid to be tested does not contain a novel coronavirus antibody, so that the liquid to be tested cannot be combined with the europium-containing time-resolved fluorescence microsphere marked sheep anti-human IgM antibody on the combination pad to form an immune complex, the europium-containing time-resolved fluorescence microsphere marked sheep anti-human IgM antibody can not be combined with the novel coronavirus NP antigen through the detection line, thus the red reaction line is not displayed at the detection line, and the europium-containing time-resolved fluorescence microsphere marked sheep anti-human IgM antibody can be combined with the rabbit anti-sheep IgM antibody through the quality control line, thus the red reaction line is displayed at the quality control line;
3. If the red reaction line is not displayed at the quality control line on the nitrocellulose membrane, the measurement is needed again as an invalid experimental result.
It should be noted that, the above-mentioned method uses a fluorescent torch with excitation wavelength of 365nm to irradiate the nitrocellulose membrane to judge the result, and can also use a dry fluorescent detector to detect the fluorescent value judging result at the detection line and the quality control line, specifically as follows: if the fluorescence values of the detection line and the quality control line are both greater than 50, the liquid to be detected contains a novel coronavirus antibody, namely the person to be detected is infected with the novel coronavirus; if the fluorescence value at the quality control line is greater than 50 and the fluorescence value at the detection line is less than 50, the liquid to be tested does not contain the novel coronavirus antibody, i.e. the person to be tested is not infected with the novel coronavirus; if the fluorescence value at the quality control line is below 50, the test result is invalid and needs to be re-measured.
Optimization of fluorescent immunochromatographic test papers for detecting novel coronavirus antibodies prepared in example 2 and example 1
The inventors of the present invention optimized the conjugate pad, and specifically optimized the boric acid buffer solution when preparing the conjugate pad.
1. Optimization of pH of boric acid buffer
1. 25MM borate buffer (pH 6.0), 25mM borate buffer (pH 6.5), 25mM borate buffer (pH 7.0), 25mM borate buffer (pH 7.5), 25mM borate buffer (pH 8.0), 25mM borate buffer (pH 8.5), 25mM borate buffer (pH 9.0) and 25mM borate buffer (pH 9.5) were prepared, respectively.
2. According to the method of steps one to three of example 1, the boric acid buffer solution of step 1 was replaced with 25mM boric acid buffer solution (pH 6.0), 25mM boric acid buffer solution (pH 6.5), 25mM boric acid buffer solution (pH 7.0), 25mM boric acid buffer solution (pH 7.5), 25mM boric acid buffer solution (pH 8.0), 25mM boric acid buffer solution (pH 8.5), 25mM boric acid buffer solution (pH 9.0) and 25mM boric acid buffer solution (pH 9.5), respectively, and the other steps were unchanged, to obtain test paper 1-test paper 8 in this order.
3. Serum of a patient infected with the novel coronavirus is taken and diluted to 10 times by PBS buffer solution, so as to obtain serum diluent.
4. 100 Mu L of serum diluent is added into a sample pad (or a sample adding hole) of the test paper 1-the test paper 8 respectively for reaction for 15min, and then a fluorescent flashlight with excitation wavelength of 365nm is used for irradiating the nitrocellulose membrane, so that the condition of red reaction lines appearing at the detection line is observed.
The detection results are shown in Table 1. The results show that when the europium-containing time-resolved fluorescence microsphere-labeled sheep anti-human IgM antibody solution is prepared, the optimal pH value of the boric acid buffer solution is 8.0.
TABLE 1
| Test paper | PH value of | Red reaction line condition displayed at detection line |
| Test paper 1 | 6.0 | - |
| Test paper 2 | 6.5 | + |
| Test paper 3 | 7.0 | ++ |
| Test paper 4 | 7.5 | +++ |
| Test paper 5 | 8.0 | ++++ |
| Test paper 6 | 8.5 | ++ |
| Test paper 7 | 9.0 | - |
| Test paper 8 | 9.5 | - |
Note that: indicating that no red reaction lines are present, + indicating that there are faint bands of subtle visibility, ++ means that there is a visible band, +++ indicates that there is a clear band visible, +++ indicates that there is a possibility the clear strip can be seen in the clear.
2. Optimization of boric acid buffer concentration
1.5 MM borate buffer (pH 8.0), 10mM borate buffer (pH 8.0), 25mM borate buffer (pH 8.0), 50mM borate buffer (pH 8.0), 100mM borate buffer (pH 8.0), 200mM borate buffer (pH 8.0) and 400mM borate buffer (pH 8.0) were prepared, respectively.
2. According to the method of steps one to three of example 1, the boric acid buffer solution of step 1 was replaced with 5mM boric acid buffer solution (pH 8.0), 10mM boric acid buffer solution (pH 8.0), 25mM boric acid buffer solution (pH 8.0), 50mM boric acid buffer solution (pH 8.0), 100mM boric acid buffer solution (pH 8.0), 200mM boric acid buffer solution (pH 8.0) and 400mM boric acid buffer solution (pH 8.0), respectively, and the other steps were unchanged, to obtain test paper a-test paper g in this order.
3. Serum of a patient infected with the novel coronavirus is taken and diluted to 10 times by PBS buffer solution, so as to obtain serum diluent.
4. 100 Mu L of serum diluent is added into a sample pad (or a sample adding hole) of test paper a-test paper g respectively for reaction for 15min, and then a fluorescent flashlight with excitation wavelength of 365nm is used for irradiating the nitrocellulose membrane, so that the condition of red reaction lines appearing at the detection line is observed.
The detection results are shown in Table 2. The results showed that when a solution of europium-containing time-resolved fluorescent microsphere-labeled goat anti-human IgM antibody was prepared, the optimal concentration of boric acid buffer solution was 25mM.
TABLE 2
Note that: indicating that no red reaction lines are present, + indicating that there are faint bands of subtle visibility, ++ means that there is a visible band, +++ indicates that there is a clear band visible, +++ indicates that there is a possibility the clear strip can be seen in the clear.
The above results indicate that when the fluorescent immunochromatographic test strip for detecting novel coronavirus antibodies was prepared by the method of example 1, the optimal boric acid buffer was 25mM boric acid buffer (pH 8.0).
Example 3 accuracy test of fluorescent immunochromatographic strip for detecting novel coronavirus antibody
1. According to the method from step one to step three of example 1, the boric acid buffer solution in step one is replaced by 25mM boric acid buffer solution (pH 8.0), and other steps are unchanged, so that the optimal fluorescent immune chromatography test paper for detecting the novel coronavirus antibody is obtained.
2. 16 Clinically diagnosed sera of novel coronavirus infected persons (named G1-G16) were randomly taken and diluted to 10 times with PBS buffer solution respectively to obtain serum dilutions of infected persons.
3. Serum from 15 healthy persons (designated as J1-J15) was randomly taken and diluted 10-fold with PBS buffer, respectively, to give dilutions of healthy human serum.
4. And (3) adding 100 mu L of serum diluent (infectious agent serum diluent or healthy human serum diluent) into a sample pad (or a sample adding hole) of the fluorescent immunochromatographic test paper prepared in the step (1) for reacting for 15min, and then detecting fluorescent values at a detection line and a quality control line by adopting a dry fluorescent detector. The following judgment is carried out according to the fluorescence value: if the fluorescence values of the detection line and the quality control line are both greater than 50, the serum diluent contains novel coronavirus antibodies, namely, the to-be-detected person infects the novel coronavirus; if the fluorescence value at the quality control line is greater than 50 and the fluorescence value at the detection line is less than 50, the serum diluent does not contain the novel coronavirus antibody, i.e. the person to be tested is not infected with the novel coronavirus; if the fluorescence value at the quality control line is below 50, the test result is invalid and needs to be re-measured.
The detection results are shown in Table 3. The result shows that the fluorescent immunochromatography test paper prepared in the step 1 can detect whether a tested person is a novel coronavirus infected person, and the accuracy rate reaches 100%.
TABLE 3 Table 3
Example 4 sensitivity test of fluorescent immunochromatographic test paper for detecting novel coronavirus antibody
1. Taking serum of a patient with a clinically definite diagnosis of a novel coronavirus infection, and diluting the serum with PBS buffer solution to obtain serum diluent; the dilution factor of the serum dilution is 20 times, 40 times, 80 times, 160 times, 320 times or 640 times.
2. 100 Mu L of serum diluent is added into a sample pad (or a sample adding hole) of the fluorescent immunochromatographic test paper prepared in the step 1 in the example 3, and the reaction is carried out for 15min, and then a dry type fluorescent detector is adopted to detect fluorescent values at a detection line and a quality control line. The following judgment is carried out according to the fluorescence value: if the fluorescence values of the detection line and the quality control line are both greater than 50, the blood serum diluent contains novel coronavirus antibodies, namely the to-be-detected person is infected with the novel coronavirus; if the fluorescence value at the quality control line is greater than 50 and the fluorescence value at the detection line is less than 50, the serum diluent does not contain the novel coronavirus antibody, i.e. the to-be-tested person is not infected with the novel coronavirus; if the fluorescence value at the quality control line is below 50, the test result is an ineffective test result and needs to be re-measured.
The detection results are shown in Table 4. The results show that when the serum dilution of the novel coronavirus infected person is 640 times, the fluorescence values at the detection line and the quality control line are still greater than 50. Therefore, the fluorescent immunity chromatographic test paper for detecting the novel coronavirus antibody has higher sensitivity.
TABLE 4 Table 4
Example 5 specificity experiment of fluorescent immunochromatographic test paper for detecting novel coronavirus antibody
1. Randomly taking 5 clinically diagnosed sera of influenza A virus infected persons (named L1-L5), and respectively diluting the sera with PBS buffer solution to 10 times to obtain serum dilutions of the influenza A virus infected persons.
2. Randomly taking 5 clinically diagnosed sera of influenza B virus infected persons (named as Y1-Y5), and respectively diluting the sera with PBS buffer solution to 10 times to obtain the serum diluent of the influenza B virus infected persons.
3. 10 Healthy human serum (named J21-J30) was randomly taken and diluted 10-fold with PBS buffer, respectively, to obtain a healthy human serum dilution.
4. Randomly taking 5 clinically diagnosed sera of new coronavirus infected persons (named X1-X5), and respectively diluting the sera with PBS buffer solution to 10 times to obtain new coronavirus infected person serum dilutions.
5. 100 Mu L of serum diluent (serum diluent of a patient suffering from influenza A, B, healthy people or new crown) is added into the sample pad (or sample adding hole) of the fluorescent immunochromatographic test paper prepared in the step 1 in the example 3, and the reaction is carried out for 15min, and then a dry fluorescent detector is adopted to detect fluorescent values at a detection line and a quality control line. The following judgment is carried out according to the fluorescence value: if the fluorescence values of the detection line and the quality control line are both greater than 50, the serum diluent contains novel coronavirus antibodies, namely the to-be-detected person is infected with the novel coronavirus; if the fluorescence value at the quality control line is greater than 50 and the fluorescence value at the detection line is less than 50, the serum diluent does not contain the novel coronavirus antibody, i.e. the to-be-tested person is not infected with the novel coronavirus; if the fluorescence value at the quality control line is below 50, the test result is invalid and needs to be re-measured.
The detection results are shown in Table 5. The results show that the fluorescence values at the detection line and the quality control line are still greater than 50 when only serum of a novel coronavirus infected person is added. Therefore, the fluorescence immunochromatography test paper for detecting the novel coronavirus antibody has higher specificity.
TABLE 5
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Claims (1)
1. A fluorescent immunochromatographic test paper for detecting novel coronavirus antibodies, the preparation method thereof is as follows:
(1) The preparation method of the bonding pad comprises the following specific steps:
(1-1) BSA was dissolved in a boric acid buffer solution (pH 8.0, 25 mM) to obtain a BSA working solution 1 having a concentration of 0.3% and a BSA working solution 2 having a concentration of 0.1%;
(1-2) after the step (1-1) is completed, adding goat anti-human IgM antibody into the europium-containing time-resolved fluorescence microsphere, uniformly mixing for 60min, then adding BSA working solution 1, and blocking overnight;
(1-3) after the step (1-2) is completed, centrifuging, collecting precipitate and re-suspending with BSA working solution 2 to obtain a europium-containing time-resolved fluorescence microsphere-labeled goat anti-human IgM antibody solution;
(1-4) spraying the europium-containing time-resolved fluorescence microsphere-labeled sheep anti-human IgM antibody solution prepared in the step (1-3) on a glass cellulose membrane, and drying to obtain a bonding pad;
(2) The preparation method comprises the following specific steps of:
(2-1) diluting the novel coronavirus NP antigen with 0.01M PBS buffer at pH7.4 to obtain a detection line solution having a concentration of 3mg/mL for coating the detection line; diluting rabbit anti-sheep IgM antibody with 0.01M PBS buffer solution with pH of 7.4 to obtain quality control line solution with concentration of 2mg/mL for coating quality control line;
(2-2) uniformly spraying the quality control line solution and the detection line solution obtained in the step (2-1) on a nitrocellulose membrane respectively, wherein the spraying amount is 10 mu L/cm 2, forming a quality control line and a detection line which are mutually separated, and drying at 37 ℃ for 3-5 hours to obtain the nitrocellulose membrane coated with the quality control line and the detection line; the quality control line and the detection line are coated on the nitrocellulose membrane, and the distance between the quality control line and the detection line is 0.5cm;
(3) The water absorption pad is stuck to one end of a nitrocellulose membrane coated with a quality control line and a detection line by double faced adhesive tape, and the overlapping length is 0.2cm; then, sticking one end of a bonding pad on the other end of the nitrocellulose membrane coated with the quality control line and the detection line by using double-sided adhesive, wherein the overlapping length is 0.2cm; and (3) sticking the sample pad on the other end of the bonding pad by using double-sided adhesive tape, wherein the overlapping length is 0.2cm, and finally sticking the sample pad and the double-sided adhesive tape on a plastic backboard, and cutting into the fluorescent immunochromatography test paper with the width of 3mm for detecting the novel coronavirus antibody.
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Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106248927A (en) * | 2016-07-21 | 2016-12-21 | 上海奥普生物医药有限公司 | The time-resolved fluoroimmunoassay chromatography reagent of a kind of Quantitative detection CK MB and preparation method |
| CN108132347A (en) * | 2018-02-09 | 2018-06-08 | 河南省生物工程技术研究中心有限公司 | The time-resolved fluoroimmunoassay chromatograph test strip and kit of joint-detection CA19-9 and CEA |
| CN109580946A (en) * | 2018-12-11 | 2019-04-05 | 深圳市亿立方生物技术有限公司 | Test strips and preparation method a kind of while that detect A type and influenza B virus |
| CN110850093A (en) * | 2019-11-22 | 2020-02-28 | 佛山墨赛生物技术有限公司 | Test strip for rapidly and quantitatively detecting ketamine in hair, preparation method and kit |
| CN111273017A (en) * | 2020-03-02 | 2020-06-12 | 江苏扬新生物医药有限公司 | Fluorescence immunochromatography kit for rapidly detecting novel coronavirus |
| CN111351941A (en) * | 2020-02-21 | 2020-06-30 | 南京岚煜生物科技有限公司 | Novel coronavirus IgM detection reagent, reagent card, kit and preparation method thereof |
| CN111426842A (en) * | 2020-02-21 | 2020-07-17 | 南京岚煜生物科技有限公司 | Novel coronavirus IgM/IgG detection reagent, reagent card, kit and preparation method thereof |
| CN111426844A (en) * | 2020-03-13 | 2020-07-17 | 南京农业大学 | Novel fluorescence immunochromatographic test strip for combined detection of coronavirus SARS-CoV-2 IgG-IgM antibody |
| CN111426829A (en) * | 2020-03-13 | 2020-07-17 | 南京农业大学 | Quantum dot microsphere immunochromatography test strip for detecting total amount of SARS-CoV-2 IgM-IgG antibody |
| CN111458505A (en) * | 2020-04-07 | 2020-07-28 | 浙江理工大学绍兴生物医药研究院有限公司 | Detection paper and method for simultaneously and quantitatively detecting IgG and IgM contents of new coronavirus |
-
2020
- 2020-08-25 CN CN202010862067.1A patent/CN111999507B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106248927A (en) * | 2016-07-21 | 2016-12-21 | 上海奥普生物医药有限公司 | The time-resolved fluoroimmunoassay chromatography reagent of a kind of Quantitative detection CK MB and preparation method |
| CN108132347A (en) * | 2018-02-09 | 2018-06-08 | 河南省生物工程技术研究中心有限公司 | The time-resolved fluoroimmunoassay chromatograph test strip and kit of joint-detection CA19-9 and CEA |
| CN109580946A (en) * | 2018-12-11 | 2019-04-05 | 深圳市亿立方生物技术有限公司 | Test strips and preparation method a kind of while that detect A type and influenza B virus |
| CN110850093A (en) * | 2019-11-22 | 2020-02-28 | 佛山墨赛生物技术有限公司 | Test strip for rapidly and quantitatively detecting ketamine in hair, preparation method and kit |
| CN111351941A (en) * | 2020-02-21 | 2020-06-30 | 南京岚煜生物科技有限公司 | Novel coronavirus IgM detection reagent, reagent card, kit and preparation method thereof |
| CN111426842A (en) * | 2020-02-21 | 2020-07-17 | 南京岚煜生物科技有限公司 | Novel coronavirus IgM/IgG detection reagent, reagent card, kit and preparation method thereof |
| CN111273017A (en) * | 2020-03-02 | 2020-06-12 | 江苏扬新生物医药有限公司 | Fluorescence immunochromatography kit for rapidly detecting novel coronavirus |
| CN111426844A (en) * | 2020-03-13 | 2020-07-17 | 南京农业大学 | Novel fluorescence immunochromatographic test strip for combined detection of coronavirus SARS-CoV-2 IgG-IgM antibody |
| CN111426829A (en) * | 2020-03-13 | 2020-07-17 | 南京农业大学 | Quantum dot microsphere immunochromatography test strip for detecting total amount of SARS-CoV-2 IgM-IgG antibody |
| CN111458505A (en) * | 2020-04-07 | 2020-07-28 | 浙江理工大学绍兴生物医药研究院有限公司 | Detection paper and method for simultaneously and quantitatively detecting IgG and IgM contents of new coronavirus |
Non-Patent Citations (1)
| Title |
|---|
| Chunyan Zhang et al.Establishing a high sensitivity detection method for SARS-CoV-2 IgM/IgG and developing a clinical application of this method.《Emerging Microbes & Infections》.2020,9(1),摘要，第2021-2023页，图1. * |
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