CN112341526A - Novel coronavirus nucleocapsid protein specific antigen polypeptide and application thereof - Google Patents
Novel coronavirus nucleocapsid protein specific antigen polypeptide and application thereof Download PDFInfo
- Publication number
- CN112341526A CN112341526A CN202011013668.1A CN202011013668A CN112341526A CN 112341526 A CN112341526 A CN 112341526A CN 202011013668 A CN202011013668 A CN 202011013668A CN 112341526 A CN112341526 A CN 112341526A
- Authority
- CN
- China
- Prior art keywords
- novel coronavirus
- antigen polypeptide
- nucleocapsid protein
- specific antigen
- kit
- 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
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 23
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 23
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 23
- 239000000427 antigen Substances 0.000 title claims abstract description 17
- 102000036639 antigens Human genes 0.000 title claims abstract description 17
- 108091007433 antigens Proteins 0.000 title claims abstract description 17
- 108700002099 Coronavirus Nucleocapsid Proteins Proteins 0.000 title claims abstract description 16
- 241000711573 Coronaviridae Species 0.000 claims abstract description 17
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 210000002966 serum Anatomy 0.000 abstract description 13
- 241001678559 COVID-19 virus Species 0.000 abstract description 6
- 208000015181 infectious disease Diseases 0.000 abstract description 4
- 208000025721 COVID-19 Diseases 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 102000004190 Enzymes Human genes 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 6
- 150000001413 amino acids Chemical group 0.000 description 6
- 239000000523 sample Substances 0.000 description 6
- 238000002835 absorbance Methods 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- 108090001074 Nucleocapsid Proteins Proteins 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 244000309467 Human Coronavirus Species 0.000 description 3
- 101710141454 Nucleoprotein Proteins 0.000 description 3
- 206010057190 Respiratory tract infections Diseases 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 239000013642 negative control Substances 0.000 description 3
- 241000008904 Betacoronavirus Species 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 2
- 241000711467 Human coronavirus 229E Species 0.000 description 2
- 241001109669 Human coronavirus HKU1 Species 0.000 description 2
- 241000482741 Human coronavirus NL63 Species 0.000 description 2
- 241001428935 Human coronavirus OC43 Species 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 230000000890 antigenic effect Effects 0.000 description 2
- 210000003719 b-lymphocyte Anatomy 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 229910000397 disodium phosphate Inorganic materials 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 230000000241 respiratory effect Effects 0.000 description 2
- 238000003757 reverse transcription PCR Methods 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 description 1
- FSBCNCKIQZZASN-GUBZILKMSA-N Ala-Arg-Met Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(O)=O FSBCNCKIQZZASN-GUBZILKMSA-N 0.000 description 1
- RTZCUEHYUQZIDE-WHFBIAKZSA-N Ala-Ser-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RTZCUEHYUQZIDE-WHFBIAKZSA-N 0.000 description 1
- 241000004176 Alphacoronavirus Species 0.000 description 1
- HPNDKUOLNRVRAY-BIIVOSGPSA-N Asn-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CC(=O)N)N)C(=O)O HPNDKUOLNRVRAY-BIIVOSGPSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 208000001528 Coronaviridae Infections Diseases 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 208000005577 Gastroenteritis Diseases 0.000 description 1
- GDOZQTNZPCUARW-YFKPBYRVSA-N Gly-Gly-Glu Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CCC(O)=O GDOZQTNZPCUARW-YFKPBYRVSA-N 0.000 description 1
- 101710114810 Glycoprotein Proteins 0.000 description 1
- 206010022678 Intestinal infections Diseases 0.000 description 1
- 241000127282 Middle East respiratory syndrome-related coronavirus Species 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 101000933967 Pseudomonas phage KPP25 Major capsid protein Proteins 0.000 description 1
- 208000004756 Respiratory Insufficiency Diseases 0.000 description 1
- 108091006197 SARS-CoV-2 Nucleocapsid Protein Proteins 0.000 description 1
- 241001678561 Sarbecovirus Species 0.000 description 1
- 101710147732 Small envelope protein Proteins 0.000 description 1
- 101710167605 Spike glycoprotein Proteins 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 208000012873 acute gastroenteritis Diseases 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000012470 diluted sample Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 108010067216 glycyl-glycyl-glycine Proteins 0.000 description 1
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 108010016686 methionyl-alanyl-serine Proteins 0.000 description 1
- 238000002887 multiple sequence alignment Methods 0.000 description 1
- 201000009240 nasopharyngitis Diseases 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000007918 pathogenicity Effects 0.000 description 1
- 238000013081 phylogenetic analysis Methods 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 201000004193 respiratory failure Diseases 0.000 description 1
- 208000020029 respiratory tract infectious disease Diseases 0.000 description 1
- 230000000405 serological effect Effects 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- CDBYLPFSWZWCQE-UHFFFAOYSA-L sodium carbonate Substances [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000012089 stop solution Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 108091005703 transmembrane proteins Proteins 0.000 description 1
- 102000035160 transmembrane proteins Human genes 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 239000011534 wash buffer Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
- C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA 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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/20011—Coronaviridae
- C12N2770/20022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Virology (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Gastroenterology & Hepatology (AREA)
- Food Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Peptides Or Proteins (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a novel coronavirus nucleocapsid protein specific antigen polypeptide and application thereof; the amino acid sequence of the antigen polypeptide is shown as SEQ ID NO.1 or SEQ ID NO. 2. The novel coronavirus nucleocapsid protein specific antigen polypeptide can be used for preparing a kit for detecting a novel coronavirus specific antibody, and the kit can detect a patient serum SARS-CoV-2 specific antibody. The kit has high specificity, good repeatability and simple and quick operation, and can be widely used for evaluating the infection condition of the novel coronavirus.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a novel coronavirus nucleocapsid protein specific antigen polypeptide and application thereof.
Background
The new type of coronavirus Disease (Corona Virus Disease 2019, COVID-19) is caused by infection with a new type of coronavirus (SARS-CoV-2). The virus is susceptible to all humans.
In the phylogenetic classification, coronaviruses (CoV) belong to the order nestvirales, the family coronaviridae, the subfamily orthocoronaviruses. The family is divided into four coronavirus genera of alpha, beta, gamma and delta according to genome characteristics[5]. At present, the WHO confirms seven human coronaviruses (HCoVs) in total, belonging to alpha-CoV (HCoV-229E and HCoV-NL63) and beta-CoV (HCoV-OC43, HCoV-HKU1, MERS-CoV, SARS-CoV-2), respectively. Human coronary viruses can cause respiratory and intestinal infections in humans, such as common cold in adults, upper respiratory infections in children, and acute gastroenteritis in infants and newborns, and severe individuals can develop acute respiratory syndrome, respiratory failure, and even death. The four human coronaviruses HCoV-229E, HCoV-OC43, HCoV-HKU1 and HCoV-NL63 are weak in pathogenicity, persist in the human population, are distributed in various regions of the world, and often cause respiratory tract infection in winter and early spring. SARS-CoV-2 belongs to the genus beta coronavirus, subgenus Sarbecovirus, based on phylogenetic analysis. Structurally, the coronavirus is spherical or elliptic, has diameter of 60-220nm, and has envelope with spinous processes on the surface and radial arrangement. The inside of the viral particle is composed of RNA and nucleocapsid protein. The viral nucleic acid is single-stranded positive-strand RNA, 27-33kb in length, is not fragmented, and is the longest of known RNA viruses. Coronaviruses contain four major structural proteins: nucleocapsid protein (N), transmembrane protein (M), small envelope protein (E) and spike glycoprotein (S). The N protein is a structural protein of coronavirus, combines with virus RNA to form nucleocapsid, and is a main antigen molecule. The N protein is abundantly expressed during viral infection and induces a strong immune response, often as a diagnostic target for coronavirus infection, and is used to assess the seroprevalence of different coronaviruses in human populations.
The development of a rapid diagnosis COVID-19 serological detection method has important significance for evaluating and controlling the epidemic of the infectious diseases. At present, COVID-19 patients are mainly diagnosed by reverse transcription polymerase chain reaction (RT-PCR) in laboratory diseases, but SARS-CoV-2 needs to be operated in a laboratory with biosafety level 2 or more, operators need to be trained professionally, and separation and identification need longer time.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a novel coronavirus nucleocapsid protein specific antigen polypeptide and application thereof.
The purpose of the invention is realized by the following technical scheme: a novel coronavirus nucleocapsid protein specific antigen polypeptide, wherein the amino acid sequence of the antigen polypeptide is shown as SEQ ID NO.1: NSPARMASGGGET or SEQ ID NO.2: TSPARMAGNGGDA, respectively.
An application of the novel coronavirus nucleocapsid protein specific antigen polypeptide in preparing a kit for detecting a novel coronavirus specific antibody.
The invention has the beneficial effects that the polypeptide-enzyme-linked immunosorbent assay kit for detecting the specific antibody of the novel coronavirus nucleocapsid protein is constructed by utilizing the antigen polypeptide, so that the infection condition of the novel coronavirus can be quickly and effectively evaluated. The kit can detect SARS-CoV-2 specific antibody in serum of a patient. The kit has high specificity, good repeatability and simple, convenient and quick operation, and can be widely used for evaluating the infection condition of the novel coronavirus.
Drawings
FIG. 1 is a graph of the predicted results of linear epitopes of the novel coronavirus nucleocapsid protein, wherein a and b are predicted epitope sites;
FIG. 2 is a diagram showing an alignment of amino acid sequences of 2 predicted antigenic polypeptides in the human coronavirus nucleocapsid protein;
FIG. 3 is a graph showing the analysis of the results of detecting antibodies specific to the serum nucleocapsid protein of a patient having COVID-19 using the kit of the present invention.
Detailed Description
Example 1
The amino acid sequence of SARS-Cov-2N protein (GenBank accession number YP-009724397) was analyzed for antigenicity, hydrophilicity, and surface accessibility by DNAStar Lasergene's Protean. Selecting amino acid segments with antigenicity index not less than 0, hydrophilicity index not less than 0 and protein surface accessibility index not less than 1, comprehensively analyzing the results, selecting polypeptide segments with all 2 factors being accorded, judging the polypeptide segments as potential linear B cell epitope, obtaining 2 segments of polypeptide in total, and respectively locating 25-37(a) and 205-217(B) segments (table 1) of each prediction site at the N end.
Table 1: prediction of linear B cell epitopes of novel coronavirus N protein
Is a weighted average of the indices corresponding to each amino acid of the polypeptide sequence.
Example 2
The new coronavirus N protein was subjected to multiple sequence alignments with the amino acid sequences of the N proteins of 6 other human-infected coronaviruses using ClustalX2.1 software, and as a result, the two antigenic polypeptides predicted to have specificity in example 1 were obtained (FIG. 2).
Example 3
1. Preparing a pre-coated enzyme standard plate: artificially synthesizing antigen polypeptides SEQ ID NO.1: NSPARMASGGGET and SEQ ID NO.2: TSPARMAGNGGDA, and dissolving and diluting the polypeptides to 10 mu g/mL by using a coating buffer solution; adding 100 mu L of enzyme label plate with 96 holes, reacting for 12h at 4 ℃, and washing the plate for 3 times by using buffer solution; adding 200 μ L of blocking solution into each well, incubating at 37 deg.C for 1h, washing plate for 3 times, freeze drying, sealing with sealing film, and storing at 4 deg.C.
2. Preparation of a reagent:
(1) coating buffer (pH 9.6): so as to contain 1.59g/L of Na2CO3And 2.93g/L NaHCO3An aqueous solution of (a).
(2) PBS buffer (pH 7.4): 8.0g NaCl, 0.2g KH2PO4、2.9g Na2HPO4 .12H2O and 0.2g KCl in 1000mL distilled water.
(3) Blocking solution, sample diluent and enzyme diluent: 1g of bovine serum albumin was dissolved in 100mL of PBS buffer.
(4) Enzyme-labeled antibody at working concentration: mu.L of a commercially available horseradish peroxidase-labeled goat anti-human IgG antibody was dissolved in 5000. mu.L of the enzyme dilution.
(5) 25-fold concentrated washing solution: 6g/L KH2PO4、5g/L KCl、36g/L Na2HPO4200g/L NaCl and 1.25% Tween 20 by volume fraction, and diluting with purified water 25 times before use to obtain washing buffer solution for use.
(6) Positive control serum: blood from patients with COVID-19 was obtained, separated to obtain serum, and diluted 20-fold.
(7) Negative control serum: blood of normal people is obtained, serum is obtained by separation, and the serum is diluted by 20 times.
(8) The TMB color development liquid and the stop solution adopt commercial reagents, such as a Kangji TMB substrate color development kit.
And (3) forming a detection kit by the pre-coated enzyme standard plate prepared in the step (1) and the reagent prepared in the step (2).
3. Sample detection: the sample was diluted 20-fold with sample diluent. Add 100. mu.L of pre-diluted sample to each well of the pre-coated plate, and set up negative, positive and blank wells simultaneously, incubate for 1 hour at 37 ℃. The plate was washed 5 times with washing solution. mu.L of horseradish peroxidase-labeled goat anti-human IgG (1:5000) was added to each well and incubated at 37 ℃ for 30 min. The plate was washed 5 times with washing solution. Adding substrate TMB developing solution for 15min, adding 50 μ L2M sulfuric acid to terminate reaction, and detecting absorbance A (A) at 450nm with microplate reader450). And judging the result by using a P/N ratio method, wherein the sample serum absorbance value/negative control absorbance value is positive when being more than or equal to 2.1, and the sample serum absorbance value/negative control absorbance value is negative when not being more than or equal to 2.1.
Example 4
Serum was obtained from 1 patient with COVID-19 and 1 normal person at 3/23 days 2020. Using the detection kit of the present invention, the detection was performed in the same manner as in example 3, and all the results were expressed as mean. + -. standard deviation, and the obtained results are shown in FIG. 3, in which OD is negative serum OD of normal person450< Cutoff value, while patient serum OD450Cutoff value.
Sequence listing
<110> Hangzhou college of medicine
<120> novel coronavirus nucleocapsid protein specific antigen polypeptide and application thereof
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 1
Asn Ser Pro Ala Arg Met Ala Ser Gly Gly Gly Glu Thr
1 5 10
<210> 2
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 2
Thr Ser Pro Ala Arg Met Ala Gly Asn Gly Gly Asp Ala
1 5 10
Claims (2)
1. A novel coronavirus nucleocapsid protein specific antigen polypeptide, wherein the amino acid sequence of the antigen polypeptide is shown as SEQ ID NO.1 or SEQ ID NO. 2.
2. Use of the novel coronavirus nucleocapsid protein-specific antigen polypeptide of claim 1 in the preparation of a kit for detecting a novel coronavirus-specific antibody.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011013668.1A CN112341526A (en) | 2020-09-24 | 2020-09-24 | Novel coronavirus nucleocapsid protein specific antigen polypeptide and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011013668.1A CN112341526A (en) | 2020-09-24 | 2020-09-24 | Novel coronavirus nucleocapsid protein specific antigen polypeptide and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112341526A true CN112341526A (en) | 2021-02-09 |
Family
ID=74358083
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011013668.1A Pending CN112341526A (en) | 2020-09-24 | 2020-09-24 | Novel coronavirus nucleocapsid protein specific antigen polypeptide and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112341526A (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005012337A2 (en) * | 2003-07-15 | 2005-02-10 | Crucell Holland B.V. | Antigenic peptides of sars coronavirus and uses thereof |
| WO2005018538A2 (en) * | 2003-06-04 | 2005-03-03 | Vaxim, Inc. | Severe acute respiratory syndrome (sars) polypeptides, antibodies to sars polypeptides and the use thereof in diagnostic, vaccination and therapeutic applications |
| CN1609119A (en) * | 2003-10-23 | 2005-04-27 | 中国医学科学院药物研究所 | Peptide library, its synthesis process and active segment screened from the peptide library |
| WO2005103259A1 (en) * | 2004-04-26 | 2005-11-03 | University Health Network | Sars-cov nucleocapsid protein epitopes and uses thereof |
| CN1826356A (en) * | 2003-07-22 | 2006-08-30 | 克鲁塞尔荷兰公司 | Binding molecules against sars-coronavirus and uses thereof |
| US20090017069A1 (en) * | 2000-06-29 | 2009-01-15 | Lipid Sciences, Inc. | SARS Vaccine Compositions and Methods of Making and Using Them |
| CN111089962A (en) * | 2020-03-25 | 2020-05-01 | 中山生物工程有限公司 | Colloidal gold kit for joint detection of novel coronavirus IgM/IgG antibody and preparation method thereof |
| CN111239392A (en) * | 2020-02-26 | 2020-06-05 | 浙江诺迦生物科技有限公司 | Novel coronavirus pneumonia (COVID-19) serological diagnosis kit |
| CN111308072A (en) * | 2020-03-25 | 2020-06-19 | 中山生物工程有限公司 | Colloidal gold immunochromatography kit for rapidly detecting novel coronavirus IgG antibody and preparation method thereof |
-
2020
- 2020-09-24 CN CN202011013668.1A patent/CN112341526A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090017069A1 (en) * | 2000-06-29 | 2009-01-15 | Lipid Sciences, Inc. | SARS Vaccine Compositions and Methods of Making and Using Them |
| WO2005018538A2 (en) * | 2003-06-04 | 2005-03-03 | Vaxim, Inc. | Severe acute respiratory syndrome (sars) polypeptides, antibodies to sars polypeptides and the use thereof in diagnostic, vaccination and therapeutic applications |
| WO2005012337A2 (en) * | 2003-07-15 | 2005-02-10 | Crucell Holland B.V. | Antigenic peptides of sars coronavirus and uses thereof |
| CN1826356A (en) * | 2003-07-22 | 2006-08-30 | 克鲁塞尔荷兰公司 | Binding molecules against sars-coronavirus and uses thereof |
| CN1609119A (en) * | 2003-10-23 | 2005-04-27 | 中国医学科学院药物研究所 | Peptide library, its synthesis process and active segment screened from the peptide library |
| WO2005103259A1 (en) * | 2004-04-26 | 2005-11-03 | University Health Network | Sars-cov nucleocapsid protein epitopes and uses thereof |
| CN111239392A (en) * | 2020-02-26 | 2020-06-05 | 浙江诺迦生物科技有限公司 | Novel coronavirus pneumonia (COVID-19) serological diagnosis kit |
| CN111089962A (en) * | 2020-03-25 | 2020-05-01 | 中山生物工程有限公司 | Colloidal gold kit for joint detection of novel coronavirus IgM/IgG antibody and preparation method thereof |
| CN111308072A (en) * | 2020-03-25 | 2020-06-19 | 中山生物工程有限公司 | Colloidal gold immunochromatography kit for rapidly detecting novel coronavirus IgG antibody and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| MAOHUA LI ET AL: "Generation of antibodies against COVID-19 virus for development of diagnostic tools", 《MEDRXIV》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ripperger et al. | Orthogonal SARS-CoV-2 serological assays enable surveillance of low-prevalence communities and reveal durable humoral immunity | |
| Sheikhzadeh et al. | Diagnostic techniques for COVID-19 and new developments | |
| CN111693712B (en) | Method for detecting SARS-CoV-2N protein of new coronavirus by using nucleic acid aptamer | |
| CN113009153B (en) | New coronavirus neutralizing antibody detection kit based on magnetic particle chemiluminescence and application thereof | |
| CN111426840A (en) | Novel coronavirus detection test strip and preparation method and application thereof | |
| CN113009154A (en) | One-step method novel magnetic microsphere detection kit for coronavirus neutralizing antibody and application thereof | |
| WO2021169664A1 (en) | Antigen for 2019 novel coronavirus and detection use thereof | |
| Liu et al. | Comparative research on nucleocapsid and spike glycoprotein as the rapid immunodetection targets of COVID-19 and establishment of immunoassay strips | |
| US20210263031A1 (en) | Immunoassay methods and compositions for detecting infection involving use of test antigens as cross-reactive control antigens | |
| TW201300421A (en) | Reagents and methods for PRRSV detection | |
| CN112462061A (en) | Kit for detecting H1N1, RSV-A and ADV3 and application thereof | |
| KR102648716B1 (en) | Antigen protein composition for diagnosing infection of African Swan Fever and use thereof | |
| EP4154007A1 (en) | Method for inactivating sars-cov-2 and its use for detecting antibodies | |
| WO2023040026A1 (en) | Immunochromatographic detection reagent strip and kit comprising same, and applications of both | |
| CN113238056A (en) | Method for detecting cystic echinococcosis | |
| CN108956986B (en) | Newcastle disease virus antibody detection kit | |
| CN112341526A (en) | Novel coronavirus nucleocapsid protein specific antigen polypeptide and application thereof | |
| Direksin et al. | An immunoperoxidase monolayer assay for the detection of antibodies against swine influenza virus | |
| EP2023142A1 (en) | Immunoassay utilizing recombinant nucleocapsid-proteins for detection of antibodies to human coronaviruses | |
| CN112912394B (en) | Monoclonal antibodies directed against the N antigen of HRSV useful for the treatment of infections, detection and diagnosis thereof | |
| JP6357425B2 (en) | Interfering peptide and method for detecting microorganisms | |
| TWI767434B (en) | Protein microarray, detection method thereof, use thereof and kit containing the same | |
| CN116519944B (en) | Indirect ELISA (enzyme-Linked immuno sorbent assay) detection method for newcastle disease virus antibody and kit thereof | |
| CN113624965B (en) | Application of N-protein specific IgG4 in screening novel coronavirus infected person and vaccinated person | |
| CN112444626B (en) | African swine fever virus antibody ELISA detection kit and preparation method thereof |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20210209 |
|
| WD01 | Invention patent application deemed withdrawn after publication |