CN111272721A - Novel coronavirus fluorescent quantitative rapid detection kit - Google Patents
Novel coronavirus fluorescent quantitative rapid detection kit Download PDFInfo
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- CN111272721A CN111272721A CN202010185370.2A CN202010185370A CN111272721A CN 111272721 A CN111272721 A CN 111272721A CN 202010185370 A CN202010185370 A CN 202010185370A CN 111272721 A CN111272721 A CN 111272721A
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- novel coronavirus
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- sars
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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- 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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks
Abstract
The invention relates to a novel coronavirus (2019-nCoV) fluorescent quantitative rapid detection kit. The kit is characterized in that fluorescent microspheres (202 nm) are used as antibody markers, and a test line T line and a control line C line are shared. The test paper confirms the positive of the novel coronavirus by diagnosing the positive of SARS-CoV NP protein, is convenient and quick to use, and is suitable for quickly diagnosing diseases caused by the novel coronavirus in various medical institutions.
Description
Technical Field
The invention belongs to the technical field of immunology in-vitro diagnosis products in the technical field of biology.
Background
The 2019-nCoV is similar to SARS coronavirus and MERS coronavirus, which are SARS coronavirus and respiratory syndrome in middle east of 2003, belonging to β genus coronavirus, in the experiment, through the gene sequence alignment of the three viruses, the 2019-nCoV has 85% similarity with SARS-CoV and 40% similarity with MERS-CoV.
The immunological diagnostic method of 2019-nCoV is similar to SARS-CoV and MERS-CoV, and mainly depends on two structural proteins encoded by the genome, namely spike protein (S) and Nucleocapsid Protein (NP).
Therefore, the SARS-CoV NP recombinant protein can be applied to the rapid diagnosis and detection of coronavirus immunology.
Disclosure of Invention
The invention provides a novel coronavirus (2019-nCoV) fluorescent quantitative rapid detection kit. The invention confirms the positive of the novel coronavirus by simultaneously diagnosing the positive of the SARS-CoV NP protein, is convenient and quick to use, and is suitable for quickly diagnosing diseases caused by the novel coronavirus in various medical institutions.
The invention adopts the following technical scheme:
a novel coronavirus (2019-nCoV) fluorescent quantitative rapid detection kit adopts fluorescent microspheres (202 nm) as antibody markers, and has a test line T and a control line C. The kit confirms the positive diagnosis of the novel coronavirus by diagnosing the positive of SARS-CoV NP protein.
Further, the SARS-CoV NP protein antibody marker is fluorescent microsphere (202 nm).
Further, the T-line is coated with a monoclonal antibody to SARS-CoV NP.
Further, the fluorescent microsphere is marked by a monoclonal antibody which can recognize SARS-CoV NP.
The invention provides a novel coronavirus (2019-nCoV) fluorescent quantitative rapid detection kit, which is used for determining the positivity of the novel coronavirus by diagnosing the protein positivity of SARS-CoV NP, is convenient and rapid to use, and is suitable for rapid diagnosis of diseases caused by the novel coronavirus in various medical institutions.
The test paper prepared by the invention has the characteristics of high sensitivity, short detection time and high accuracy.
Description of the drawings:
FIG. 1 is a schematic diagram of a fluorescent quantitative rapid detection kit for a novel coronavirus (2019-nCoV).
The specific implementation mode is as follows:
for the purpose of making the objects, technical solutions and advantages of the present invention more apparent, the present invention will be further described in detail with reference to the accompanying drawings and embodiments, it being understood that the specific embodiments described herein are for the purpose of illustration only and are not to be construed as limiting the invention.
Preparation of the test paper:
the first embodiment is as follows:
preparing 0.02M PB solution, adding 0.05% Tween 20 and 0.01% bovine serum albumin, soaking a sample pad of 300mm x 150mm for 30min, and drying at 60 ℃ for later use.
Coating liquid is prepared, 0.1 percent of sucrose and 0.2 percent of bovine serum albumin are added into 0.02M PB solution.
Preparing a complex solution, adding 0.5% of sucrose, 0.1% of Tween 20 and 0.5% of bovine serum albumin into 0.01M PB solution.
Coating goat anti-mouse IgG and SARS-CoV NP monoclonal antibody each 10-30ul, scribing C, T, drying for use.
Marking 10-50ul of SARS-CoV NP monoclonal antibody with 202nm fluorescent microsphere, subpackaging into 96-hole non-adsorption enzyme label plate, and freeze-drying for use.
And assembling the fluorescent test strip, namely sequentially assembling the base plate, the nc membrane, the absorbent paper and the sample pad, and cutting the base plate, the nc membrane, the absorbent paper and the sample pad into strips.
The second embodiment is as follows:
preparing 0.02M tris solution, adding 0.05% Tween 20 and 0.01% bovine serum albumin, soaking a sample pad of 300mm x 150mm for 30min, and drying at 60 ℃ for later use.
Coating liquid is prepared, 0.02M tris solution is added with 0.1 percent of sucrose and 0.2 percent of bovine serum albumin.
Preparing a complex solution, 0.01M tris solution, and adding 0.5% of sucrose, 0.1% of Tween 20 and 0.5% of bovine serum albumin.
Coating goat anti-mouse IgG and SARS-CoV NP monoclonal antibody each 10-30ul, scribing C, T, drying for use.
Marking 10-50ul of SARS-CoV NP monoclonal antibody with 202nm fluorescent microsphere, subpackaging into 96-hole non-adsorption enzyme label plate, and freeze-drying for use.
And assembling the fluorescent test strip, namely sequentially assembling the base plate, the nc membrane, the absorbent paper and the sample pad, and cutting the base plate, the nc membrane, the absorbent paper and the sample pad into strips.
The use method of the test paper comprises the following steps:
diluting the quality control product or sample to 50 times, sucking 50ul into the freeze-dried fluorescent marker hole, blowing and beating uniformly, reacting for 5min, sucking the liquid in the hole and dripping onto the sample pad of the test strip, waiting for 10min, and then putting into a fluorescent reading instrument to read data, as shown in figure 1.
The test paper prepared by the invention has the characteristics of high sensitivity, short detection time and high accuracy, and the kit is a novel coronavirus (2019-nCoV) fluorescent quantitative rapid detection kit.
Claims (4)
1. A novel coronavirus fluorescent quantitative rapid detection kit is characterized in that: the detection kit adopts fluorescent microspheres (202 nm) as antibody markers, has a test line T line and a control line C line, and confirms the positive diagnosis of the novel coronavirus by diagnosing the positive of SARS-CoV NP protein.
2. The kit for fluorescent quantitative rapid detection of coronavirus according to claim 1, wherein: the SARS-CoV NP protein antibody marker adopts fluorescent microspheres (202 nm).
3. The kit for fluorescent quantitative rapid detection of coronavirus according to claim 1, wherein: the T line is coated with a monoclonal antibody to SARS-CoV NP.
4. The kit for fluorescent quantitative rapid detection of coronavirus according to claim 1, wherein: the fluorescent microsphere is marked by a monoclonal antibody for recognizing SARS-CoV NP.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010185370.2A CN111272721A (en) | 2020-03-17 | 2020-03-17 | Novel coronavirus fluorescent quantitative rapid detection kit |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010185370.2A CN111272721A (en) | 2020-03-17 | 2020-03-17 | Novel coronavirus fluorescent quantitative rapid detection kit |
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| CN111272721A true CN111272721A (en) | 2020-06-12 |
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| CN202010185370.2A Pending CN111272721A (en) | 2020-03-17 | 2020-03-17 | Novel coronavirus fluorescent quantitative rapid detection kit |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114539393A (en) * | 2020-11-25 | 2022-05-27 | 北京纽安博新逸生物科技有限公司 | 2019-novel coronavirus N protein single domain antibody and application thereof |
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| CN106872420A (en) * | 2016-12-27 | 2017-06-20 | 厦门奥德生物科技有限公司 | The kit and method of a kind of time-resolved fluorescence quantitative determination microdose urine protein |
| CN106957247A (en) * | 2017-03-16 | 2017-07-18 | 长春万成生物电子工程有限公司 | A kind of purification process of micro protein indicator |
| CN107942061A (en) * | 2017-11-29 | 2018-04-20 | 洛阳现代生物技术研究院有限公司 | A kind of test card, preparation and its detection method for detecting transmissible gastro-enteritis virus antibody |
| CN108982828A (en) * | 2018-06-28 | 2018-12-11 | 长春万成生物电子工程有限公司 | A kind of quantum dot examines the preparation and application of test paper fastly |
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2020
- 2020-03-17 CN CN202010185370.2A patent/CN111272721A/en active Pending
Patent Citations (5)
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| CN102539766A (en) * | 2010-12-29 | 2012-07-04 | 北京宝瑞源科技孵化有限公司 | Beta-bungarotoxin detection kit and preparation method thereof |
| CN106872420A (en) * | 2016-12-27 | 2017-06-20 | 厦门奥德生物科技有限公司 | The kit and method of a kind of time-resolved fluorescence quantitative determination microdose urine protein |
| CN106957247A (en) * | 2017-03-16 | 2017-07-18 | 长春万成生物电子工程有限公司 | A kind of purification process of micro protein indicator |
| CN107942061A (en) * | 2017-11-29 | 2018-04-20 | 洛阳现代生物技术研究院有限公司 | A kind of test card, preparation and its detection method for detecting transmissible gastro-enteritis virus antibody |
| CN108982828A (en) * | 2018-06-28 | 2018-12-11 | 长春万成生物电子工程有限公司 | A kind of quantum dot examines the preparation and application of test paper fastly |
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| BO DIAO ET AL: "Human Kidney is a Target for Novel Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Infection", 《MEDRXIV》, 4 March 2020 (2020-03-04), pages 1 - 13 * |
| XINTIAN XU ET AL: "Evolution of the novel coronavirus from the ongoing Wuhan outbreak and modeling of its spike protein for risk of human transmission", 《SCIENCE CHINA LIFE SCIENCES》, 21 January 2020 (2020-01-21), pages 457 - 460 * |
| 何长民等: "《生物制品基础》", 31 July 1986, 甘肃人民出版社, pages: 369 - 370 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114539393A (en) * | 2020-11-25 | 2022-05-27 | 北京纽安博新逸生物科技有限公司 | 2019-novel coronavirus N protein single domain antibody and application thereof |
| CN114539393B (en) * | 2020-11-25 | 2023-06-09 | 北京纽安博新逸生物科技有限公司 | 2019-novel coronavirus N protein single domain antibody and application thereof |
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Application publication date: 20200612 |