CN111398599A - Novel coronavirus IgG/IgM antibody detection kit and application method thereof - Google Patents
Novel coronavirus IgG/IgM antibody detection kit and application method thereof Download PDFInfo
- Publication number
- CN111398599A CN111398599A CN202010217588.1A CN202010217588A CN111398599A CN 111398599 A CN111398599 A CN 111398599A CN 202010217588 A CN202010217588 A CN 202010217588A CN 111398599 A CN111398599 A CN 111398599A
- Authority
- CN
- China
- Prior art keywords
- colloidal gold
- detection
- novel coronavirus
- igg
- sample
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000001514 detection method Methods 0.000 title claims abstract description 98
- 241000711573 Coronaviridae Species 0.000 title claims abstract description 59
- 238000000034 method Methods 0.000 title claims abstract description 18
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 62
- 102000036639 antigens Human genes 0.000 claims abstract description 33
- 108091007433 antigens Proteins 0.000 claims abstract description 33
- 239000000020 Nitrocellulose Substances 0.000 claims abstract description 31
- 239000012528 membrane Substances 0.000 claims abstract description 31
- 229920001220 nitrocellulos Polymers 0.000 claims abstract description 31
- 238000006243 chemical reaction Methods 0.000 claims abstract description 16
- 230000002745 absorbent Effects 0.000 claims abstract description 11
- 239000002250 absorbent Substances 0.000 claims abstract description 11
- 239000000427 antigen Substances 0.000 claims abstract description 7
- 238000003908 quality control method Methods 0.000 claims description 24
- 241000283707 Capra Species 0.000 claims description 14
- SWGJCIMEBVHMTA-UHFFFAOYSA-K trisodium;6-oxido-4-sulfo-5-[(4-sulfonatonaphthalen-1-yl)diazenyl]naphthalene-2-sulfonate Chemical compound [Na+].[Na+].[Na+].C1=CC=C2C(N=NC3=C4C(=CC(=CC4=CC=C3O)S([O-])(=O)=O)S([O-])(=O)=O)=CC=C(S([O-])(=O)=O)C2=C1 SWGJCIMEBVHMTA-UHFFFAOYSA-K 0.000 claims description 11
- 210000004369 blood Anatomy 0.000 claims description 6
- 239000008280 blood Substances 0.000 claims description 6
- 239000003085 diluting agent Substances 0.000 claims description 5
- 239000002245 particle Substances 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- 210000002966 serum Anatomy 0.000 claims description 4
- 239000010931 gold Substances 0.000 claims description 3
- 229910052737 gold Inorganic materials 0.000 claims description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 2
- 238000003113 dilution method Methods 0.000 claims description 2
- 238000007689 inspection Methods 0.000 abstract description 3
- 238000012544 monitoring process Methods 0.000 abstract description 3
- 239000013589 supplement Substances 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 9
- 238000010586 diagram Methods 0.000 description 3
- 239000001509 sodium citrate Substances 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 3
- 229940038773 trisodium citrate Drugs 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 101001018064 Homo sapiens Lysosomal-trafficking regulator Proteins 0.000 description 2
- 102100033472 Lysosomal-trafficking regulator Human genes 0.000 description 2
- 235000010703 Modiola caroliniana Nutrition 0.000 description 2
- 244000038561 Modiola caroliniana Species 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 108010008595 sarcoma-associated antigen S1 Proteins 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- 108700002099 Coronavirus Nucleocapsid Proteins Proteins 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000004568 cement Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- 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
- G01N33/585—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
- G01N33/587—Nanoparticles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/165—Coronaviridae, e.g. avian infectious bronchitis virus
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2469/00—Immunoassays for the detection of microorganisms
- G01N2469/20—Detection of antibodies in sample from host which are directed against antigens from microorganisms
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Food Science & Technology (AREA)
- General Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Pathology (AREA)
- Virology (AREA)
- Nanotechnology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention belongs to the technical field of biological detection, and particularly relates to a novel coronavirus IgG/IgM antibody detection kit, which comprises a sample pad, a colloidal gold pad, a nitrocellulose membrane, absorbent filter paper and a reaction support; the sample pad, the colloidal gold pad, the nitrocellulose membrane and the absorbent filter paper are sequentially pasted on a reaction support, one end of the sample pad is pressed and covered with one end of the colloidal gold pad, the other end of the colloidal gold pad is pressed and covered with one end of the nitrocellulose membrane, the absorbent filter paper is pressed and covered with the other end of the nitrocellulose membrane, and the colloidal gold pad is a colloidal gold pad containing a novel coronavirus S1 recombinant antigen protein, an N recombinant antigen protein and a mouse IgG antigen protein which are marked by colloidal gold. The invention also provides an application method of the detection kit. The invention can be used as a supplement for the detection of novel coronavirus antigens, thereby reducing the missing detection; the method has the advantages of no dependence on any experimental instrument, environment or personnel, simple and convenient operation, short detection period, easy interpretation and convenient monitoring and inspection at any time and any place.
Description
Technical Field
The invention belongs to the technical field of biological detection, and particularly relates to a novel coronavirus IgG/IgM antibody detection kit and an application method thereof.
Background
At present, the existing novel coronavirus detection kit has the problems of long detection time consumption, complex operation steps and the like. Therefore, the development of a kit for rapidly detecting the novel coronavirus is of great significance for controlling the epidemic situation of the novel coronavirus.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a novel coronavirus IgG/IgM antibody detection kit and an application method thereof, and aims to solve the technical problems that the existing novel coronavirus detection kit is long in detection time consumption, complicated in operation steps and low in detection efficiency of new coronaviruses.
The invention provides a novel coronavirus IgG/IgM antibody detection kit, which has the following specific technical scheme:
the novel coronavirus IgG/IgM antibody detection kit comprises a sample pad, a colloidal gold pad, a nitrocellulose membrane, absorbent filter paper and a reaction support;
the sample pad, the colloidal gold pad, the nitrocellulose membrane and the water-absorbing filter paper are sequentially stuck on the reaction support, one end of the sample pad is pressed and covered on one end of the colloidal gold pad, the other end of the colloidal gold pad is pressed and covered on one end of the nitrocellulose membrane, the water-absorbing filter paper is pressed and covered on the other end of the nitrocellulose membrane,
the colloidal gold pad is a colloidal gold pad containing a novel coronavirus S1 recombinant antigen protein, an N recombinant antigen protein and a mouse IgG antigen protein which are marked by colloidal gold.
In some embodiments, the nitrocellulose membrane is provided with a first detection strip, a second detection strip and a quality control strip, wherein the first detection strip and the second detection strip are positioned on one side of the nitrocellulose membrane close to the colloidal gold pad, the first detection strip contains an anti-human IgG monoclonal antibody, the second detection strip contains an anti-human IgM monoclonal antibody, and the quality control strip is positioned on one side of the nitrocellulose membrane away from the colloidal gold pad and contains a goat anti-mouse IgG polyclonal antibody.
In some embodiments, the first detection strip, the second detection strip and the quality control strip are each 0.35-0.4cm in length and 0.5-1mm in width, and the spacing between the first detection strip and the second detection strip and the spacing between the second detection strip and the quality control strip are each 3.5-4 mm.
In certain embodiments, the diameter of the gold particles in the antigen-bound gold colloid is 20 to 50 nm.
In some embodiments, the material of the reaction support is plastic.
The invention also provides an application method of the novel coronavirus IgG/IgM antibody detection kit, which is based on the novel coronavirus IgG/IgM antibody detection kit and comprises the following steps:
s1, carrying out diluent treatment on the sample to be detected to obtain a treated sample;
s2, dripping the processed sample in the step S1 onto the sample pad, wherein the processed sample moves to the direction of the absorbent filter paper along each attachment on the reaction support in turn;
s3, if the processed sample contains novel coronavirus IgG antibodies and IgM antibodies, in the process of passing through the colloidal gold pad, the novel coronavirus S1 recombinant antigen protein and N recombinant antigen protein which are marked by the colloidal gold pad are specifically combined to form a complex; when the complex passes through the nitrocellulose membrane, the complex is specifically combined with the anti-human IgG monoclonal antibody and the anti-human IgM monoclonal antibody in sequence, so that red or purple red appears at the first detection strip and the second detection strip, and the sample to be detected is positive; if the treated sample does not contain the IgG/IgM antibody of the novel coronavirus, the first detection strip and the second detection strip are not red or purple red, and the sample is negative; the colloidal gold labeled mouse IgG antigen protein continues to advance to be combined with the goat anti-mouse IgG polyclonal antibody, and the quality control strip appears red or purple red;
s4, if the quality control strip appears red or purple red, the result shows that the colloidal gold labeled mouse IgG antigen protein is normally combined with the goat anti-mouse IgG polyclonal antibody, and the experimental result is effective; if the red or purple red does not appear at the quality control band, the surface gelatin body is invalid or the experimental operation is wrong, and the experimental result is invalid.
In some embodiments, in step S1, the sample to be tested is serum, plasma or whole blood; the sample diluent in the dilution process is normal saline.
The invention has the following beneficial effects: infection with new corona causes the body to mount an immune response and produce virus-specific antibodies, which are detected in a blood sample (serum, plasma or whole blood). The novel coronavirus IgG/IgM antibody detection kit provided by the invention can simplify the detection process of novel coronaviruses. The detection kit provided by the invention is used for detecting the novel coronavirus IgG/IgM antibody, has no dependence on any experimental instrument, environment or operator, and is simple and convenient to operate, short in detection period and easy to interpret; in addition, special instruments and equipment are not needed, professional training is not needed, adaptability is strong, and monitoring and inspection at any time and any place are facilitated. The application method of the novel coronavirus IgG/IgM antibody detection kit provided by the invention is simple and convenient to operate, does not depend on higher laboratory conditions, is used for quickly detecting the novel coronavirus IgG/IgM antibody, and improves the detection efficiency of the novel coronavirus.
Drawings
FIG. 1 is a front view of the novel coronavirus IgG/IgM antibody detection kit of the present invention;
FIG. 2 is a side view of the novel coronavirus IgG/IgM antibody detection kit of the present invention;
FIG. 3 is a schematic diagram showing the positive detection result of the novel coronavirus IgG/IgM antibody detection kit of the present invention;
FIG. 4 is a schematic diagram showing the negative result of the detection by the novel coronavirus IgG/IgM antibody detection kit of the present invention;
FIG. 5 is a schematic diagram showing the results of the novel coronavirus IgG/IgM antibody detection kit of the present invention are invalid.
Detailed Description
In order that the objects, aspects and advantages of the present invention will become more apparent, the present invention will be further described in detail with reference to the following detailed description of preferred embodiments thereof, with reference to the accompanying drawings of fig. 1-5.
The embodiment of the invention is a specific detection mode aiming at novel coronavirus IgG/IgM antibody, and detection methods of other subtypes can be obtained according to the following ideas after appropriate antigens are prepared.
The novel coronavirus IgG/IgM antibody detection kit provided by the invention consists of water-absorbing filter paper 1, a nitrocellulose membrane 2, a colloidal gold pad 3, a sample pad 4 and a reaction support 5. The sample pad 4, the colloidal gold pad 3, the nitrocellulose membrane 2 and the water-absorbing filter paper 1 are sequentially stuck on the reaction support 5, one end of the sample pad 4 is pressed and covered on one end of the colloidal gold pad 3, the other end of the colloidal gold pad 3 is pressed and covered on one end of the nitrocellulose membrane 2, and the water-absorbing filter paper 1 is pressed and covered on the other end of the nitrocellulose membrane 2. Preferably, the reaction support 5 may be a plastic cement plate. The absorbent filter paper 1 can prevent the sample from being accumulated at one end of the nitrocellulose membrane 2, so that the subsequent sample cannot move forward continuously, and the reaction can be fully performed. Preferably, the nitrocellulose membrane 2 includes a first detection band G and a second detection band M containing an anti-human IgG monoclonal antibody and an anti-human IgM monoclonal antibody, and a quality control band C containing a goat anti-mouse IgG polyclonal antibody. The colloidal gold pad 3 contains a novel coronavirus S1 recombinant antigen protein, an N recombinant antigen protein and a mouse IgG antigen protein which are labeled by colloidal gold. The nitrocellulose membrane 2 provided with a first detection strip G, a second detection strip M, and a quality control strip C is stuck on the reaction support 5.
Preferably, the lengths of the first detection strip G, the second detection strip M and the quality control strip C are 0.35-0.4cm, the widths of the first detection strip G, the second detection strip M and the quality control strip C are 0.5-1mm, and the distance between the first detection strip G and the second detection strip M and the distance between the second detection strip M and the quality control strip C are 3.5-4 mm.
Preferably, in the novel coronavirus IgG/IgM antibody detection kit of the present invention, the diameter of colloidal gold particles in the antigen-bound colloidal gold in the binder region is 20 to 50 nm. The colloidal gold may be colloidal gold prepared by a method conventional in the art, and for example, the method of preparing colloidal gold may be trisodium citrate reduction.
The colloidal gold for antigen binding is prepared by adopting a trisodium citrate reduction method, taking a 0.01% chloroauric acid aqueous solution 100m L by mass fraction, heating to boiling, quickly adding 1% trisodium citrate with 1.75m L, reacting for 10min, cooling, pouring the solution into a clean glass ware, measuring the maximum absorption peak value of the prepared colloidal gold by a spectrophotometer, and storing at 4 ℃ for later use.
In addition, the novel coronavirus IgG/IgM antibody detection kit of the present invention may be prepared by coating a colloidal gold solution containing an antigen bound thereto on the nitrocellulose membrane 2.
The preparation method of the novel coronavirus IgG/IgM antibody detection kit provided by the invention comprises the following specific steps:
(1) adding 2m L potassium carbonate solution (0.1 mol/L) into colloidal gold solution (100m L), uniformly stirring, adding 1mg of novel coronavirus S1 antigen protein, reacting for 15min, adding 10% bovine serum albumin solution 10m L, sealing for 10min, transferring the antibody-labeled colloidal gold solution into a centrifuge tube, centrifuging at 4 ℃ of 10000 r/min and 10cm at the centrifugal radius for 30min, removing supernatant, adding 3m L PBS (0.02 mol/L containing 1% bovine serum albumin) solution with the pH of 8.2 to redissolve and precipitate, and storing at 4 ℃ for later use, and carrying out colloidal gold labeling on the novel coronavirus N antigen protein and the mouse IgG protein by the same method, wherein the diameter of colloidal gold particles in the colloidal gold solution is 20-50 nm;
(3) attaching a sample pad 4, a conjugate pad, a nitrocellulose membrane 2 and absorbent paper to a support plate in a manner of overlapping one another in sequence to form a test paper substrate;
(4) g, M and a line C are coated on the nitrocellulose membrane 2 on the paper substrate by a three-dimensional plane film scribing instrument, wherein the line C is goat anti-mouse IgG polyclonal antibody with the concentration of 1.5mg/M L, the line G is mouse anti-human IgG monoclonal antibody, the line M is mouse anti-human IgM monoclonal antibody with the concentration of 1mg/M L;
(5) and cutting the test paper substrate into test paper strips with the width of 3.5-4 mm.
In order to detect the novel coronavirus IgG/IgM antibodies, a suitable diluent formulation is selected, and in this example, physiological saline is used as the sample treatment solution.
50-100 ul of sample treatment solution to be detected is directly dripped into the sample pad 4 of the detection kit, at the moment, the sample moves to the direction of the water absorption filter paper 1 along each attachment on the reaction support 5 in sequence, and the experimental result is read within 15 minutes.
If the sample to be detected (serum, plasma or whole blood) contains the novel coronavirus IgG/IgM antibody, the antibody is specifically combined with the novel coronavirus S1 recombinant antigen protein and the novel coronavirus N1 recombinant antigen protein which are labeled by colloidal gold on the detection kit respectively to form a compound, and then the compound is continuously carried forward and is specifically combined with the anti-human IgG monoclonal antibody and the anti-human IgM monoclonal antibody which are coated on the nitrocellulose membrane 2, red or mauve appears at the first detection strip G and the second detection strip M, namely a detection line, which indicates that the sample to be detected contains the novel coronavirus IgG/IgM antibody and is positive, while the mouse IgG antigen protein labeled by colloidal gold is continuously carried forward and is normally combined with the goat anti-mouse IgG polyclonal antibody, and red or mauve appears at the quality control strip C, refer to fig. 3.
If the sample to be detected does not contain the novel coronavirus IgG/IgM antibody, a sufficient complex cannot be formed, the first detection strip G and the second detection strip M cannot be red or purple, and are negative, the colloidal gold-labeled mouse IgG antigen protein is normally combined with the goat anti-mouse IgG polyclonal antibody, and the quality control strip C is red or purple, as shown in FIG. 4.
No matter whether the sample contains the novel coronavirus IgG/IgM antibody or not, the mouse IgG antigen protein marked by the colloidal gold can continuously move to the quality control strip C and is combined with the goat anti-mouse IgG polyclonal antibody coated at the position to form a red or purple red strip, namely a quality control line. If the detection result does not appear in the quality control line, the colloidal gold is proved to be invalid or the operation is failed, the result is invalid, and the re-detection is needed, and the reference is made to FIG. 5.
In this embodiment, the novel coronavirus recombinant S1 antigen protein and the recombinant N antigen protein are produced by near-shore protein technologies limited; the goat anti-human IgG monoclonal antibody, the goat anti-human IgM monoclonal antibody, the mouse IgG and the goat anti-mouse IgG polyclonal antibody are produced by the Fenpeng biological corporation;
the novel coronavirus IgG/IgM antibody detection kit provided by the invention can simplify the detection process of the novel coronavirus IgG/IgM antibody in a blood sample. The detection kit is used for detecting the novel coronavirus IgG/IgM antibody, does not have dependence on any experimental instrument, environment or operator, and is simple and convenient to operate, short in detection period and easy to interpret; in addition, special instruments and equipment are not needed, professional training is not needed, the adaptability is strong, and the monitoring and the inspection at any time and any place are convenient.
The above description is only for the purpose of illustrating preferred embodiments of the present invention and is not to be construed as limiting the invention, and the present invention is not limited to the above examples, and those skilled in the art should also be able to make various changes, modifications, additions or substitutions within the spirit and scope of the present invention.
Claims (7)
1. The novel coronavirus IgG/IgM antibody detection kit is characterized by comprising a sample pad, a colloidal gold pad, a nitrocellulose membrane, absorbent filter paper and a reaction support;
the sample pad, the colloidal gold pad, the nitrocellulose membrane and the absorbent filter paper are sequentially stuck on the reaction support, one end of the sample pad is pressed and covered on one end of the colloidal gold pad, the other end of the colloidal gold pad is pressed and covered on one end of the nitrocellulose membrane, the absorbent filter paper is pressed and covered on the other end of the nitrocellulose membrane,
the colloidal gold pad is a colloidal gold pad containing a novel coronavirus S1 recombinant antigen protein, an N recombinant antigen protein and a mouse IgG antigen protein which are marked by colloidal gold.
2. The novel coronavirus IgG/IgM antibody detection kit according to claim 1, wherein the nitrocellulose membrane is provided with a first detection strip, a second detection strip and a quality control strip, the first detection strip and the second detection strip are positioned on one side of the nitrocellulose membrane close to the colloidal gold pad, the first detection strip contains an anti-human IgG monoclonal antibody, the second detection strip contains an anti-human IgM monoclonal antibody, and the quality control strip is positioned on one side of the nitrocellulose membrane away from the colloidal gold pad and contains a goat anti-mouse IgG polyclonal antibody.
3. The novel coronavirus IgG/IgM antibody detection kit according to claim 2, wherein the first detection strip, the second detection strip and the quality control strip are each 0.35-0.4cm in length and 0.5-1mm in width, and the distance between the first detection strip and the second detection strip and the distance between the second detection strip and the quality control strip are each 3.5-4 mm.
4. The novel coronavirus IgG/IgM antibody detection kit according to claim 1, wherein the diameter of the gold particles in the antigen-bound gold is 20 to 50 nm.
5. The kit of claim 1, wherein the reaction support is made of plastic.
6. The application method of the novel coronavirus IgG/IgM antibody detection kit is based on the novel coronavirus IgG/IgM antibody detection kit of any one of claims 1 to 5, and is characterized by comprising the following steps:
s1, carrying out diluent treatment on the sample to be detected to obtain a treated sample;
s2, dripping the processed sample in the step S1 onto the sample pad, wherein the processed sample moves to the direction of the absorbent filter paper along each attachment on the reaction support in turn;
s3, if the processed sample contains novel coronavirus IgG antibodies and IgM antibodies, in the process of passing through the colloidal gold pad, the novel coronavirus S1 recombinant antigen protein and N recombinant antigen protein which are marked by the colloidal gold pad are specifically combined to form a complex; when the complex passes through the nitrocellulose membrane, the complex is specifically combined with the anti-human IgG monoclonal antibody and the anti-human IgM monoclonal antibody in sequence, so that the first detection strip and the second detection strip are red or purple red, and the sample to be detected is positive; if the treated sample does not contain the IgG/IgM antibody of the novel coronavirus, the first detection strip and the second detection strip are not red or purple red, and the sample is negative; the colloidal gold labeled mouse IgG antigen protein continues to advance to be combined with the goat anti-mouse IgG polyclonal antibody, and the quality control strip appears red or purple red;
s4, if the quality control strip appears red or purple red, the result shows that the colloidal gold labeled mouse IgG antigen protein is normally combined with the goat anti-mouse IgG polyclonal antibody, and the experimental result is effective; if the red or purple red does not appear at the quality control band, the colloidal gold is invalid or the experimental operation is wrong, and the experimental result is invalid.
7. The method of claim 6, wherein in step S1, the sample to be tested is serum, plasma or whole blood; the sample diluent in the dilution process is normal saline.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010217588.1A CN111398599A (en) | 2020-03-25 | 2020-03-25 | Novel coronavirus IgG/IgM antibody detection kit and application method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010217588.1A CN111398599A (en) | 2020-03-25 | 2020-03-25 | Novel coronavirus IgG/IgM antibody detection kit and application method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111398599A true CN111398599A (en) | 2020-07-10 |
Family
ID=71429189
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010217588.1A Pending CN111398599A (en) | 2020-03-25 | 2020-03-25 | Novel coronavirus IgG/IgM antibody detection kit and application method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111398599A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111693714A (en) * | 2020-06-04 | 2020-09-22 | 华东理工大学 | Novel coronavirus IgM/IgG antibody detection kit and application method thereof |
| CN111781350A (en) * | 2020-07-15 | 2020-10-16 | 桂林电子科技大学 | Colloidal gold immunochromatographic test strip for detecting novel coronavirus and preparation method thereof |
| CN112114151A (en) * | 2020-08-20 | 2020-12-22 | 深圳爱信生物技术有限公司 | 2019-nCoV IgG, IgM and IgA antibody combined detection kit and detection method thereof |
| CN113970636A (en) * | 2020-07-23 | 2022-01-25 | 湖北金鉴生物有限公司 | Test paper for qualitatively detecting novel coronavirus IgG/IgM antibody and preparation method thereof |
| CN114113587A (en) * | 2020-08-26 | 2022-03-01 | 华中科技大学 | Novel coronavirus antibody colloidal gold immunochromatography detection card and application |
Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070092938A1 (en) * | 2003-07-15 | 2007-04-26 | Temasek Life Sciences Laboratory | Diagnostics for sars virus |
| CN102707057A (en) * | 2012-05-24 | 2012-10-03 | 蓝十字生物药业（北京）有限公司 | Colloidal gold test strip for fast detecting IgG and IgM antibodies of dengue fever virus |
| CN102854316A (en) * | 2012-07-31 | 2013-01-02 | 上海博沃生物科技有限公司 | Detecting kit for hepatitis a virus IgM and IgG antibodies through colloidal gold method |
| CN103412124A (en) * | 2013-07-25 | 2013-11-27 | 北京陆桥技术有限责任公司 | Aflatoxin M1 gold label quick detectiontest card and preparation method and application thereof |
| CN104634975A (en) * | 2013-11-08 | 2015-05-20 | 北京易斯威特生物医学科技有限公司 | Detection kit and detection method of Norwalk viruses |
| CN205910196U (en) * | 2016-05-26 | 2017-01-25 | 江苏雷森生物科技有限公司 | Colloidal gold test paper card of short -term test dog coronavirus antibody |
| WO2017121001A1 (en) * | 2016-01-16 | 2017-07-20 | 深圳市华科安测信息技术有限公司 | Gold-labeled antibody test strip for use in early-stage diabetic nephropathy test |
| CN108931654A (en) * | 2018-07-05 | 2018-12-04 | 重庆早柒天生物科技股份有限公司 | It is to detect early pregnancy detection kit, its production method and the purposes of sample with urine |
| CN109596824A (en) * | 2019-01-04 | 2019-04-09 | 杭州奥泰生物技术股份有限公司 | A kind of Test paper and preparation method thereof of quick diagnosis Lyme disease |
| CN110007096A (en) * | 2019-05-06 | 2019-07-12 | 江苏硕世生物科技股份有限公司 | A kind of dengue virus IgG/IgM antibody test strip, kit and preparation method thereof |
| CN110540578A (en) * | 2019-08-28 | 2019-12-06 | 中国农业科学院上海兽医研究所(中国动物卫生与流行病学中心上海分中心) | Preparation of main antigen epitope region recombinant protein of pseudorabies virus GE gene and colloidal gold immunochromatographic test strip |
-
2020
- 2020-03-25 CN CN202010217588.1A patent/CN111398599A/en active Pending
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070092938A1 (en) * | 2003-07-15 | 2007-04-26 | Temasek Life Sciences Laboratory | Diagnostics for sars virus |
| CN102707057A (en) * | 2012-05-24 | 2012-10-03 | 蓝十字生物药业（北京）有限公司 | Colloidal gold test strip for fast detecting IgG and IgM antibodies of dengue fever virus |
| CN102854316A (en) * | 2012-07-31 | 2013-01-02 | 上海博沃生物科技有限公司 | Detecting kit for hepatitis a virus IgM and IgG antibodies through colloidal gold method |
| CN103412124A (en) * | 2013-07-25 | 2013-11-27 | 北京陆桥技术有限责任公司 | Aflatoxin M1 gold label quick detectiontest card and preparation method and application thereof |
| CN104634975A (en) * | 2013-11-08 | 2015-05-20 | 北京易斯威特生物医学科技有限公司 | Detection kit and detection method of Norwalk viruses |
| WO2017121001A1 (en) * | 2016-01-16 | 2017-07-20 | 深圳市华科安测信息技术有限公司 | Gold-labeled antibody test strip for use in early-stage diabetic nephropathy test |
| CN205910196U (en) * | 2016-05-26 | 2017-01-25 | 江苏雷森生物科技有限公司 | Colloidal gold test paper card of short -term test dog coronavirus antibody |
| CN108931654A (en) * | 2018-07-05 | 2018-12-04 | 重庆早柒天生物科技股份有限公司 | It is to detect early pregnancy detection kit, its production method and the purposes of sample with urine |
| CN109596824A (en) * | 2019-01-04 | 2019-04-09 | 杭州奥泰生物技术股份有限公司 | A kind of Test paper and preparation method thereof of quick diagnosis Lyme disease |
| CN110007096A (en) * | 2019-05-06 | 2019-07-12 | 江苏硕世生物科技股份有限公司 | A kind of dengue virus IgG/IgM antibody test strip, kit and preparation method thereof |
| CN110540578A (en) * | 2019-08-28 | 2019-12-06 | 中国农业科学院上海兽医研究所(中国动物卫生与流行病学中心上海分中心) | Preparation of main antigen epitope region recombinant protein of pseudorabies virus GE gene and colloidal gold immunochromatographic test strip |
Non-Patent Citations (12)
| Title |
|---|
| ZHENGTU LI等: "Development and Clinical Application of A Rapid IgM-IgG Combined Antibody Test for SARS-CoV-2 Infection Diagnosis", 《JOURNAL OF MEDICAL VIROLOGY》 * |
| ZHENGTU LI等: "Development and Clinical Application of A Rapid IgM-IgG Combined Antibody Test for SARS-CoV-2 Infection Diagnosis", 《JOURNAL OF MEDICAL VIROLOGY》, vol. 92, no. 9, 27 February 2020 (2020-02-27), pages 1518 - 1524 * |
| 刘彬等: "新型冠状病毒基因组结构与蛋白功能", 《微生物与感染》 * |
| 刘彬等: "新型冠状病毒基因组结构与蛋白功能", 《微生物与感染》, vol. 15, no. 01, 25 February 2020 (2020-02-25), pages 52 - 57 * |
| 唐鹏等: "化学发光和胶体金法检测新型冠状病毒特异性抗体比较及其临床意义", 《武汉大学学报(医学版)》 * |
| 唐鹏等: "化学发光和胶体金法检测新型冠状病毒特异性抗体比较及其临床意义", 《武汉大学学报(医学版)》, vol. 41, no. 04, 8 March 2020 (2020-03-08) * |
| 张亚楠: "胶体金法检测新型冠状病毒特异性IgM和IgG抗体的假阳性结果分析", 《包头医学院学报》 * |
| 张亚楠: "胶体金法检测新型冠状病毒特异性IgM和IgG抗体的假阳性结果分析", 《包头医学院学报》, vol. 36, no. 03, 15 March 2020 (2020-03-15) * |
| 邱峰等: "新型冠状病毒SARS-CoV-2的实验室检测技术", 《南方医科大学学报》 * |
| 邱峰等: "新型冠状病毒SARS-CoV-2的实验室检测技术", 《南方医科大学学报》, vol. 40, no. 02, 22 February 2020 (2020-02-22) * |
| 陈福祥等: "新型冠状病毒感染肺炎的实验诊断", 《诊断学理论与实践》 * |
| 陈福祥等: "新型冠状病毒感染肺炎的实验诊断", 《诊断学理论与实践》, vol. 19, no. 01, 25 February 2020 (2020-02-25) * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111693714A (en) * | 2020-06-04 | 2020-09-22 | 华东理工大学 | Novel coronavirus IgM/IgG antibody detection kit and application method thereof |
| CN111781350A (en) * | 2020-07-15 | 2020-10-16 | 桂林电子科技大学 | Colloidal gold immunochromatographic test strip for detecting novel coronavirus and preparation method thereof |
| CN113970636A (en) * | 2020-07-23 | 2022-01-25 | 湖北金鉴生物有限公司 | Test paper for qualitatively detecting novel coronavirus IgG/IgM antibody and preparation method thereof |
| CN113970636B (en) * | 2020-07-23 | 2024-03-15 | 湖北金鉴生物有限公司 | Test paper for qualitatively detecting novel coronavirus IgG/IgM antibody and preparation method thereof |
| CN112114151A (en) * | 2020-08-20 | 2020-12-22 | 深圳爱信生物技术有限公司 | 2019-nCoV IgG, IgM and IgA antibody combined detection kit and detection method thereof |
| CN114113587A (en) * | 2020-08-26 | 2022-03-01 | 华中科技大学 | Novel coronavirus antibody colloidal gold immunochromatography detection card and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111398599A (en) | Novel coronavirus IgG/IgM antibody detection kit and application method thereof | |
| CN111366729B (en) | Novel coronavirus COVID-19 antigen fluorescence detection kit and preparation method thereof | |
| JP5567932B2 (en) | Reagent composition for immunochromatography and measuring method using the same | |
| CN111426844A (en) | Novel fluorescence immunochromatographic test strip for combined detection of coronavirus SARS-CoV-2 IgG-IgM antibody | |
| CN111912980A (en) | Rapid combined detection device for novel coronavirus antigen and antibody in saliva and preparation method thereof | |
| CN112485436A (en) | Colloidal gold immunochromatographic test strip for detecting in-vivo neutralizing antibody of neocorona vaccine after injection and preparation method thereof | |
| CN111537720A (en) | Preparation method of novel coronavirus 2019-nCoV IgM/IgG antibody combined detection kit | |
| CN111781350A (en) | Colloidal gold immunochromatographic test strip for detecting novel coronavirus and preparation method thereof | |
| CN107907679A (en) | A kind of immuno-chromatographic test paper strip and preparation method thereof and application | |
| US20230341398A1 (en) | SARS-CoV-2 DETECTION KIT AND SARS-CoV-2 DETECTION METHOD | |
| CN112326966A (en) | Novel rapid detection kit for coronavirus antigen, and preparation method and application thereof | |
| CN111624336A (en) | Novel coronavirus IgG/IgM antibody detection kit and application method thereof | |
| CN111693714A (en) | Novel coronavirus IgM/IgG antibody detection kit and application method thereof | |
| CN112485422A (en) | Latex microsphere immunochromatography test strip based on novel coronavirus antigen and preparation method thereof | |
| CN109212203B (en) | Quantum dot immunochromatography test strip for rapidly detecting brucella antibody | |
| CN208443852U (en) | Immunofluorescence for detecting dog C reactive protein chromatographs detection card | |
| CN110954695A (en) | Norovirus GI and GII type quantum dot joint inspection test strip and preparation method and application thereof | |
| CN109946457A (en) | A kind of colloidal gold strip detecting Porcine epidemic diarrhea virus | |
| CN213023175U (en) | Coronavirus antibody test strip and reagent card | |
| CN114264819A (en) | Quantum dot nanosphere immunochromatography test strip for rapidly detecting new coronavirus | |
| CN113970636B (en) | Test paper for qualitatively detecting novel coronavirus IgG/IgM antibody and preparation method thereof | |
| JP7498556B2 (en) | Immunoassay method and immunoassay device | |
| CN116194436A (en) | Test reagent with improved specificity by suppressing false negative | |
| CN111521787A (en) | New bunyavirus IgG and IgM combined detection reagent card and preparation method thereof | |
| WO2024005055A1 (en) | Test method, test reagent, and test kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200710 |