CN111273006A - Novel coronavirus SARS-CoV-2S protein detection method - Google Patents
Novel coronavirus SARS-CoV-2S protein detection method Download PDFInfo
- Publication number
- CN111273006A CN111273006A CN202010161075.3A CN202010161075A CN111273006A CN 111273006 A CN111273006 A CN 111273006A CN 202010161075 A CN202010161075 A CN 202010161075A CN 111273006 A CN111273006 A CN 111273006A
- Authority
- CN
- China
- Prior art keywords
- protein
- cov
- ace2
- recombinant protein
- alkaline phosphatase
- 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
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
- G01N33/53—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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
-
- 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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
-
- 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
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
-
- 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
- G01N2469/10—Detection of antigens from microorganism in sample from host
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a novel method for detecting SARS-CoV-2S protein of coronavirus, which utilizes spike protein on the dependent surface of SARS-CoV-2 virus to combine with angiotensin converting enzyme 2 on the cell surface to enter into cells, the method uses the raw materials commonly used in chemiluminescence immunoassay, the raw materials and ACE2 are easily purchased from the market, the biotin mark and alkaline phosphatase mark are rapid, the method can rapidly prepare the reagent for detecting SARS-CoV-2S protein, the method compares with the fluorescence quantitative PCR result of SARS-CoV-2, and the positive coincidence rate, the negative coincidence rate and the total coincidence rate are all higher.
Description
Technical Field
The invention relates to a virus detection method, in particular to a novel detection method of coronavirus SARS-CoV-2S protein.
Background
The virus antigen detection immunological analysis method usually adopts double antibody sandwich method, taking SARS-CoV N protein antigen detection as an example, SARS-CoV N protein specific antibody is at the solid phase end, SARS-CoV N protein specific antibody is at the mark end, adding the sample and incubating, SARS-CoV N protein in the sample and SARS-CoV N protein specific antibody at the solid phase end combine to form immune complex, SARS-CoV N protein specific antibody at the mark end combine with different antigen sites on SARS-CoV N protein in the sample to form marked immune complex, and removing the uncombined enzyme conjugate and other substances by washing. The marker is detected by means of color development or luminescence, etc., so that whether the SARS-CoV N protein is contained in the matter to be detected can be judged.
The key to the detection of the antigen by the method is to prepare a specific antibody aiming at the antigen. Whether the monoclonal antibody or the polyclonal antibody is prepared, the antigen is recombined and expressed through genetic engineering, then an animal is immunized to prepare the antibody, and the prepared antibody needs to be screened out for detection with high affinity. To complete the series of tasks, it usually takes much time, material and labor. Some inherent disadvantages of the conventional double antibody sandwich method are that reagents for detecting antigen have not appeared since the outbreak of SARS-CoV-2 virus.
Disclosure of Invention
The invention provides a novel method for detecting SARS-CoV-2S protein of coronavirus, which solves the problem that the prior art can not effectively detect SARS-CoV-2S protein of coronavirus.
In order to solve the technical problem, the invention provides the following technical scheme:
a novel method for detecting SARS-CoV-2S protein of coronavirus is characterized by comprising the following steps:
s1, labeling the recombinant protein of angiotensin converting enzyme 2ACE2 with biotin ester, and coating the protein on streptavidin magnetic beads;
s2, activating ACE2, and then uniformly mixing and reacting with activated alkaline phosphatase ALP to prepare ACE2 marked by alkaline phosphatase ALP;
s3, uniformly mixing the reagent in the steps 1 and 2 with a sample to be detected, and incubating;
s4, washing, and removing the unbound enzyme conjugate and other substances;
s5, adding luminescent substrate AMPPD, catalyzing the luminescent substrate by the enzyme conjugate to emit photons, and judging whether SARS-CoV-2S protein exists in the object to be detected according to the number of the photons.
Preferably, the S1 is specifically: the angiotensin converting enzyme 2ACE2 recombinant protein and Biotin ester labeling reagent NHS-LC-Biotin are mixed uniformly at room temperature according to the proportion of 1:20(mol/mol) and react for 30min, and the ACE2 recombinant protein labeled by Biotin and streptavidin magnetic beads are mixed uniformly at the proportion of 5 mu g/mg and react at room temperature for 30min to prepare the ACE2 recombinant protein coated magnetic particles.
Preferably, the process for preparing the alkaline phosphatase ALP activated in S2 is as follows: activated alkaline phosphatase ALP was prepared by mixing (N-maleimidomethyl) cyclohexane-1-carboxylic acid succinimidyl ester SMCC and alkaline phosphatase ALP at a ratio of 1:38(w/w) at room temperature for 30 min.
Preferably, the ACE2 activation process in S2 is as follows: 2-iminosulfane hydrochloride 2-IT and ACE2 recombinant protein are mixed evenly according to the proportion of 1:12(w/w) at room temperature for 30min to prepare the activated ACE2 recombinant protein.
Preferably, the preparation process of the ALP-labeled ACE2 in S2 is as follows: the activated alkaline phosphatase and the activated ACE2 recombinant protein are mixed uniformly for 2h at room temperature according to the ratio of 1:1.2(w/w) to prepare the ACE2 recombinant protein marked by the alkaline phosphatase ALP.
Preferably, the S3 is specifically: and (3) putting 10 mu L of a sample to be detected into a reaction tube, adding 50 mu L of ACE2 recombinant protein coated magnetic particles, adding 50 mu L of ACE2 recombinant protein marked by alkaline phosphatase ALP, uniformly mixing, and reacting at 37 ℃ for 20 min.
Preferably, the S3 is specifically: adding a luminescent substrate AMPPD, catalyzing the luminescent substrate by an enzyme conjugate to emit photons, wherein the number of the photons is in direct proportion to the concentration of S protein in a sample; the SARS-CoV-2 recombinant S protein is diluted in gradient to be used as the calibrator of the detection system, and the concentration of the S protein in the sample is calculated by a calibration curve of concentration-relative luminescence value (RLU).
The invention also provides a kit of a novel coronavirus SARS-CoV-2S protein detection method, the kit comprises: magnetic particles coated by biotin ester labeled ACE2 recombinant protein, alkaline phosphatase ALP labeled ACE2 recombinant protein and luminescent substrate AMPPD.
Compared with the prior art, the invention has the following advantages:
SARS-CoV-2 virus relies on the binding of the surface spike protein (S protein) to angiotensin converting enzyme 2(ACE2) on the cell surface to gain entry into the cell, and both have previously been of high affinity. The present invention designs a method for detecting SARS-CoV-2S protein by using said characteristic.
The method uses the raw materials commonly used in chemiluminescence immunoassay, and the raw materials and ACE2 are easily purchased from the market. And biotin labeling and alkaline phosphatase labeling were rapid. Can rapidly prepare a reagent for detecting SARS-CoV-2S protein.
Compared with the result of fluorescence quantitative PCR of SARS-CoV-2, the method has higher positive coincidence rate, negative coincidence rate and total coincidence rate.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following examples, and the exemplary embodiments and descriptions thereof are only used for explaining the present invention and are not used as limitations of the present invention.
Example 1
S1, mixing the recombinant protein of angiotensin converting enzyme 2ACE2 and Biotin ester labeling reagent NHS-LC-Biotin uniformly at room temperature according to the proportion of 1:20(mol/mol) and reacting for 30 min;
uniformly mixing biotin ester-labeled ACE2 recombinant protein and streptavidin magnetic beads at the ratio of 5 mu g/mg, and reacting at room temperature for 30min to prepare ACE2 recombinant protein-coated magnetic particles;
s2, mixing (N-maleimidomethyl) cyclohexane-1-carboxylic acid succinimidyl ester SMCC and alkaline phosphatase ALP at a ratio of 1:38(w/w) at room temperature for 30min to prepare activated alkaline phosphatase ALP;
uniformly mixing the 2-IT and ACE2 recombinant proteins at room temperature for 30min according to the proportion of 1:12(w/w) to prepare activated ACE2 recombinant proteins;
uniformly mixing the activated alkaline phosphatase and the activated ACE2 recombinant protein at room temperature for 2h according to the ratio of 1:1.2(w/w) to prepare the ACE2 recombinant protein marked by alkaline phosphatase ALP;
s3, putting 10 mu L of a sample to be detected into a reaction tube, adding 50 mu L of ACE2 recombinant protein coated magnetic particles, adding 50 mu L of ACE2 recombinant protein marked by alkaline phosphatase ALP, mixing uniformly, and reacting for 20min at 37 ℃;
s4, washing, and removing the unbound enzyme conjugate and other substances.
S5, adding a luminescent substrate AMPPD, and catalyzing the luminescent substrate to emit photons by the enzyme conjugate, wherein the number of the photons is in direct proportion to the concentration of the S protein in the sample. The SARS-CoV-2 recombinant S protein is diluted in gradient to be used as the calibrator of the detection system, and the concentration of the S protein in the sample is calculated by a calibration curve of concentration-relative luminescence value (RLU).
Example 2
A kit for a novel method for detecting a SARS-CoV-2S protein of a coronavirus, the kit comprising: the kit comprises magnetic particles coated by biotin ester labeled ACE2 recombinant protein, alkaline phosphatase ALP labeled ACE2 recombinant protein and a luminescent substrate AMPPD. The preparation process of the magnetic particle coated by the biotin ester labeled ACE2 recombinant protein comprises the following steps: evenly mixing angiotensin converting enzyme 2ACE2 recombinant protein and Biotin ester labeling reagent NHS-LC-Biotin according to a ratio of 1:20(mol/mol) at room temperature for reaction for 30min, evenly mixing Biotin-labeled ACE2 recombinant protein and streptavidin magnetic beads at a ratio of 5 mu g/mg at room temperature for reaction for 30min to prepare ACE2 recombinant protein coated magnetic particles; the preparation process of the ACE2 recombinant protein marked by alkaline phosphatase ALP comprises the following steps: mixing (N-maleimidomethyl) cyclohexane-1-carboxylic acid succinimidyl ester SMCC and alkaline phosphatase ALP at a ratio of 1:38(w/w) at room temperature for 30min to prepare activated alkaline phosphatase ALP; uniformly mixing 2-IT and ACE2 recombinant protein according to a ratio of 1:12(w/w) at room temperature for 30min to prepare activated ACE2 recombinant protein; and uniformly mixing the activated alkaline phosphatase and the activated ACE2 recombinant protein at room temperature for 2h according to the ratio of 1:1.2(w/w) to prepare the ACE2 recombinant protein marked by the alkaline phosphatase ALP.
The above-mentioned embodiments, objects, technical solutions and advantages of the present invention are further described in detail, it should be understood that the above-mentioned embodiments are only exemplary embodiments of the present invention, and are not intended to limit the scope of the present invention, and any modifications, equivalent substitutions, improvements and the like made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (8)
1. A novel method for detecting SARS-CoV-2S protein of coronavirus is characterized by comprising the following steps:
s1, labeling the recombinant protein of angiotensin converting enzyme 2ACE2 with biotin ester, and coating the protein on streptavidin magnetic beads;
s2, activating ACE2, and then uniformly mixing and reacting with activated alkaline phosphatase ALP to prepare ACE2 marked by alkaline phosphatase ALP;
s3, uniformly mixing the reagent in the steps 1 and 2 with a sample to be detected, and incubating;
s4, washing, and removing the unbound enzyme conjugate and other substances;
s5, adding luminescent substrate AMPPD, catalyzing the luminescent substrate by the enzyme conjugate to emit photons, and judging whether SARS-CoV-2S protein exists in the object to be detected according to the number of the photons.
2. The method for detecting SARS-CoV-2S protein as claimed in claim 1, wherein S1 is specifically: the angiotensin converting enzyme 2ACE2 recombinant protein and Biotin ester labeling reagent NHS-LC-Biotin are mixed uniformly at room temperature according to the proportion of 1:20(mol/mol) and react for 30min, and the ACE2 recombinant protein labeled by Biotin and streptavidin magnetic beads are mixed uniformly at the proportion of 5 mu g/mg and react at room temperature for 30min to prepare the ACE2 recombinant protein coated magnetic particles.
3. The method for detecting SARS-CoV-2S protein of coronavirus as claimed in claim 1, wherein the ALP activated alkaline phosphatase in S2 is prepared by: activated alkaline phosphatase ALP was prepared by mixing (N-maleimidomethyl) cyclohexane-1-carboxylic acid succinimidyl ester SMCC and alkaline phosphatase ALP at a ratio of 1:38(w/w) at room temperature for 30 min.
4. The method for detecting SARS-CoV-2S coronavirus protein as claimed in claim 1, wherein the ACE2 activation process in S2 is as follows: 2-iminosulfane hydrochloride 2-IT and ACE2 recombinant protein are mixed evenly according to the proportion of 1:12(w/w) at room temperature for 30min to prepare the activated ACE2 recombinant protein.
5. The method for detecting SARS-CoV-2S protein of coronavirus as claimed in any one of claims 1, 3 or 4, wherein the ALP-labeled ACE2 in S2 is prepared by: the activated alkaline phosphatase and the activated ACE2 recombinant protein are mixed uniformly for 2h at room temperature according to the ratio of 1:1.2(w/w) to prepare the ACE2 recombinant protein marked by the alkaline phosphatase ALP.
6. The method for detecting SARS-CoV-2S protein as claimed in claim 1, wherein S3 is specifically: and (3) putting 10 mu L of a sample to be detected into a reaction tube, adding 50 mu L of ACE2 recombinant protein coated magnetic particles, adding 50 mu L of ACE2 recombinant protein marked by alkaline phosphatase ALP, uniformly mixing, and reacting at 37 ℃ for 20 min.
7. The method for detecting SARS-CoV-2S protein as claimed in claim 1, wherein S3 is specifically: adding a luminescent substrate AMPPD, catalyzing the luminescent substrate by an enzyme conjugate to emit photons, wherein the number of the photons is in direct proportion to the concentration of S protein in a sample; the SARS-CoV-2 recombinant S protein is diluted in gradient to be used as the calibrator of the detection system, and the concentration of the S protein in the sample is calculated by a calibration curve of concentration-relative luminescence value (RLU).
8. A kit for using the novel method for detecting SARS-CoV-2S protein of coronavirus according to any one of claims 1 to 7, wherein the kit comprises: the kit comprises magnetic particles coated by biotin ester labeled ACE2 recombinant protein, alkaline phosphatase ALP labeled ACE2 recombinant protein and a luminescent substrate AMPPD.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010161075.3A CN111273006A (en) | 2020-03-10 | 2020-03-10 | Novel coronavirus SARS-CoV-2S protein detection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010161075.3A CN111273006A (en) | 2020-03-10 | 2020-03-10 | Novel coronavirus SARS-CoV-2S protein detection method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111273006A true CN111273006A (en) | 2020-06-12 |
Family
ID=71000575
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010161075.3A Pending CN111273006A (en) | 2020-03-10 | 2020-03-10 | Novel coronavirus SARS-CoV-2S protein detection method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111273006A (en) |
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112043838A (en) * | 2020-07-28 | 2020-12-08 | 北京肿瘤医院(北京大学肿瘤医院) | ACE2 receptor targeted nuclide polypeptide probe, and preparation method and application thereof |
| CN112098643A (en) * | 2020-08-20 | 2020-12-18 | 广东省农业科学院农业生物基因研究中心 | Method for evaluating cross-species infection risk of coronavirus, test strip and application of test strip |
| CN112213497A (en) * | 2020-09-24 | 2021-01-12 | 杭州医学院 | polypeptide-ELISA kit for detecting novel coronavirus S protein idiotypic antibody |
| CN112285348A (en) * | 2020-12-29 | 2021-01-29 | 北京百普赛斯生物科技股份有限公司 | Electrochemical luminescence immunoassay kit for antigen protein expressed by new coronavirus vaccine |
| CN112415189A (en) * | 2021-01-22 | 2021-02-26 | 北京百普赛斯生物科技股份有限公司 | Magnetic bead coupled with novel coronavirus S2 protein, and preparation method and application thereof |
| CN112782401A (en) * | 2021-02-08 | 2021-05-11 | 聊城大学 | Method for rapidly detecting novel coronavirus in vitro and application |
| CN113009154A (en) * | 2021-02-25 | 2021-06-22 | 山东莱博生物科技有限公司 | One-step method novel magnetic microsphere detection kit for coronavirus neutralizing antibody and application thereof |
| CN113030483A (en) * | 2021-02-25 | 2021-06-25 | 山东省大健康精准医疗产业技术研究院 | Novel coronavirus neutralizing antibody enzyme-linked immunosorbent assay detection kit and application thereof |
| CN113156119A (en) * | 2020-05-09 | 2021-07-23 | 深圳安赛诊断技术有限公司 | Method for detecting coronavirus by adopting angiotensin converting enzyme II (ACE2) |
| CN113419061A (en) * | 2020-06-19 | 2021-09-21 | 南京金斯瑞生物科技有限公司 | Magnetic particle chemiluminescence kit for detecting SARS-CoV-2 virus neutralizing antibody and application thereof |
| WO2021189058A1 (en) * | 2020-03-20 | 2021-09-23 | Victor Manneh | Coronavirus assay |
| WO2021224493A1 (en) * | 2020-05-07 | 2021-11-11 | Senseutics Limited | Method for detecting corona virus |
| WO2022003192A1 (en) * | 2020-07-02 | 2022-01-06 | Leibniz-Institut Für Photonische Technologien E.V. | Agents and method for enriching or depleting and for detecting corona viruses |
| EP4012044A1 (en) * | 2020-12-09 | 2022-06-15 | Qualizyme Diagnostics GmbH & Co KG | Method for detecting a virus |
| US11740240B2 (en) | 2020-07-20 | 2023-08-29 | Bio-Rad Laboratories, Inc. | Immunoassay for SARS-CoV-2 neutralizing antibodies and materials therefor |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103901203A (en) * | 2014-04-11 | 2014-07-02 | 苏州浩欧博生物医药有限公司 | Chemiluminescence quantitative detection kit for procalcitonin, and preparation method and detection method thereof |
| CN109182281A (en) * | 2018-08-31 | 2019-01-11 | 首都医科大学 | Handle the method and kit of adeno-associated virus |
| CN111273016A (en) * | 2020-02-26 | 2020-06-12 | 浙江诺迦生物科技有限公司 | Kit for rapidly detecting coronavirus based on S protein ligand and ACE2 receptor competition chromatography |
-
2020
- 2020-03-10 CN CN202010161075.3A patent/CN111273006A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103901203A (en) * | 2014-04-11 | 2014-07-02 | 苏州浩欧博生物医药有限公司 | Chemiluminescence quantitative detection kit for procalcitonin, and preparation method and detection method thereof |
| CN109182281A (en) * | 2018-08-31 | 2019-01-11 | 首都医科大学 | Handle the method and kit of adeno-associated virus |
| CN111273016A (en) * | 2020-02-26 | 2020-06-12 | 浙江诺迦生物科技有限公司 | Kit for rapidly detecting coronavirus based on S protein ligand and ACE2 receptor competition chromatography |
Non-Patent Citations (4)
| Title |
|---|
| ILONA GLOWACKA ET AL: "Differential Downregulation of ACE2 by the Spike Proteins of Severe Acute Respiratory Syndrome Coronavirus and Human Coronavirus NL63", 《JOURNAL OF VIROLOGY》 * |
| TIN-YUN HO ET AL: "Design and biological activities of novel inhibitory peptides for SARS-CoV spike protein and angiotensin-converting enzyme 2 interaction", 《ANTIVIRAL RES.》 * |
| XUE-FEI CAI ET AL: "A Peptide-Based Magnetic Chemiluminescence Enzyme Immunoassay for Serological Diagnosis of Coronavirus Disease 2019", 《THE JOURNAL OF INFECTIOUS DISEASES》 * |
| 孙琰等: "SARS-CoV S666蛋白与眼部ACE2受体的结合作用", 《眼科新进展》 * |
Cited By (22)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021189058A1 (en) * | 2020-03-20 | 2021-09-23 | Victor Manneh | Coronavirus assay |
| WO2021224493A1 (en) * | 2020-05-07 | 2021-11-11 | Senseutics Limited | Method for detecting corona virus |
| CN113156119A (en) * | 2020-05-09 | 2021-07-23 | 深圳安赛诊断技术有限公司 | Method for detecting coronavirus by adopting angiotensin converting enzyme II (ACE2) |
| WO2021254476A1 (en) * | 2020-06-19 | 2021-12-23 | 南京金斯瑞生物科技有限公司 | Magnetic microparticle chemiluminescence reagent kit for detecting sars-cov-2 virus neutralising antibodies and application therefor |
| CN113419061A (en) * | 2020-06-19 | 2021-09-21 | 南京金斯瑞生物科技有限公司 | Magnetic particle chemiluminescence kit for detecting SARS-CoV-2 virus neutralizing antibody and application thereof |
| WO2022003192A1 (en) * | 2020-07-02 | 2022-01-06 | Leibniz-Institut Für Photonische Technologien E.V. | Agents and method for enriching or depleting and for detecting corona viruses |
| US11740240B2 (en) | 2020-07-20 | 2023-08-29 | Bio-Rad Laboratories, Inc. | Immunoassay for SARS-CoV-2 neutralizing antibodies and materials therefor |
| CN112043838B (en) * | 2020-07-28 | 2022-07-26 | 北京肿瘤医院(北京大学肿瘤医院) | ACE2 receptor targeted nuclide polypeptide probe, and preparation method and application thereof |
| CN112043838A (en) * | 2020-07-28 | 2020-12-08 | 北京肿瘤医院(北京大学肿瘤医院) | ACE2 receptor targeted nuclide polypeptide probe, and preparation method and application thereof |
| CN112098643B (en) * | 2020-08-20 | 2022-02-15 | 广东省农业科学院农业生物基因研究中心 | Method for evaluating cross-species infection risk of coronavirus, test strip and application of test strip |
| CN112098643A (en) * | 2020-08-20 | 2020-12-18 | 广东省农业科学院农业生物基因研究中心 | Method for evaluating cross-species infection risk of coronavirus, test strip and application of test strip |
| CN112213497A (en) * | 2020-09-24 | 2021-01-12 | 杭州医学院 | polypeptide-ELISA kit for detecting novel coronavirus S protein idiotypic antibody |
| CN112213497B (en) * | 2020-09-24 | 2023-10-20 | 杭州医学院 | polypeptide-ELISA kit for detecting novel coronavirus S protein unique antibody |
| EP4012044A1 (en) * | 2020-12-09 | 2022-06-15 | Qualizyme Diagnostics GmbH & Co KG | Method for detecting a virus |
| CN112285348A (en) * | 2020-12-29 | 2021-01-29 | 北京百普赛斯生物科技股份有限公司 | Electrochemical luminescence immunoassay kit for antigen protein expressed by new coronavirus vaccine |
| CN112415189B (en) * | 2021-01-22 | 2021-03-30 | 北京百普赛斯生物科技股份有限公司 | Magnetic bead coupled with novel coronavirus S2 protein, and preparation method and application thereof |
| CN112415189A (en) * | 2021-01-22 | 2021-02-26 | 北京百普赛斯生物科技股份有限公司 | Magnetic bead coupled with novel coronavirus S2 protein, and preparation method and application thereof |
| CN112782401A (en) * | 2021-02-08 | 2021-05-11 | 聊城大学 | Method for rapidly detecting novel coronavirus in vitro and application |
| CN113030483A (en) * | 2021-02-25 | 2021-06-25 | 山东省大健康精准医疗产业技术研究院 | Novel coronavirus neutralizing antibody enzyme-linked immunosorbent assay detection kit and application thereof |
| CN113009154A (en) * | 2021-02-25 | 2021-06-22 | 山东莱博生物科技有限公司 | One-step method novel magnetic microsphere detection kit for coronavirus neutralizing antibody and application thereof |
| CN113009154B (en) * | 2021-02-25 | 2024-01-30 | 山东莱博生物科技有限公司 | Novel one-step method coronavirus neutralizing antibody magnetic microsphere detection kit and application thereof |
| CN113030483B (en) * | 2021-02-25 | 2024-04-02 | 山东省大健康精准医疗产业技术研究院 | Novel coronavirus neutralizing antibody ELISA detection kit and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111273006A (en) | Novel coronavirus SARS-CoV-2S protein detection method | |
| Porcheron et al. | Development of an enzyme immunoassay for ecdysteroids using acetylcholinesterase as label | |
| AU777123B2 (en) | Chemiluminescent assay | |
| Kricka | Ultrasensitive immunoassay techniques | |
| EP2817423B1 (en) | Methods and systems for signal amplification of bioassays | |
| WO2012122929A1 (en) | Electrochemiluminescence immunoassay method | |
| KR100905300B1 (en) | Cascade enzyme-linked immunosorbent assay | |
| US20110136256A1 (en) | Methods and kits for decreasing interferences in plasma or serum containing assay samples of specific binding assays | |
| CN112285353A (en) | Method for improving anti-biotin interference capability and sensitivity of chemiluminescence kit of streptavidin-biotin reaction system | |
| WO2023087942A1 (en) | Delisa sample capable of increasing proportion of effective microbeads, and preparation and detection method therefor | |
| CN108444988B (en) | Thyroglobulin chemiluminescence detection kit | |
| CN113640511A (en) | Magnetic particle electrochemiluminescence kit | |
| CN111693721A (en) | Preparation method and application of enzyme-linked immunosorbent assay based on prussian blue nano enzyme label | |
| EP1347301A1 (en) | Method for ultra-rapid and ultra-sensitive measurement | |
| US20080227111A1 (en) | Reagent kit and method for measuring hcv antibody | |
| CN116106559A (en) | Biotin-antibody coupling ratio detection kit and application thereof | |
| CN114152740A (en) | Enzymatic chemiluminescence kit, preparation method and detection method | |
| Härmä et al. | Immunoassay on a single microparticle: the effect of particle size and number on a miniaturized time-resolved fluorometric assay of free prostate-specific antigen | |
| CN112098651A (en) | High-sensitivity chemiluminescence immunodiagnosis reagent and method for eliminating HOOK effect of two-step method immunoreagent | |
| CN112595852A (en) | Reagent kit for detecting SARS-CoV-2 virus antibody and its preparing method | |
| WO2023182520A1 (en) | Immunological analysis method, complex, method for producing complex, and reagent for immunological analysis | |
| CN110862438B (en) | Compound and preparation method and application thereof | |
| US20220145353A1 (en) | Sensitive glucose assay | |
| CN112834742A (en) | Non-cleaning magnetic particle chemiluminescence immunoassay method | |
| CN115950877A (en) | Magnetic particle chemiluminescence kit for detecting heavy metals |
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 |
Application publication date: 20200612 |
|
| RJ01 | Rejection of invention patent application after publication |