CN111272722A - Novel fluorescent quantitative rapid detection bigeminy card kit for coronavirus - Google Patents
Novel fluorescent quantitative rapid detection bigeminy card kit for coronavirus Download PDFInfo
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- CN111272722A CN111272722A CN202010185501.7A CN202010185501A CN111272722A CN 111272722 A CN111272722 A CN 111272722A CN 202010185501 A CN202010185501 A CN 202010185501A CN 111272722 A CN111272722 A CN 111272722A
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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
- 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
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- Molecular Biology (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention relates to a novel fluorescent quantitative rapid detection bivalent card kit for coronavirus (2019-nCoV). The kit is characterized in that fluorescent microspheres (202 nm) are used as antibody markers, two cards are combined into one, and the two cards are respectively provided with a C line and a T line. The dual card can respectively diagnose SARS-CoV NP protein positive and SARS-CoV protein negative to confirm the positive of the novel coronavirus, is convenient and quick to use, and is suitable for quick diagnosis of 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 fluorescent quantitative rapid detection bigeminal kit for coronavirus (2019-nCoV), which is convenient and rapid to use and is suitable for rapid diagnosis of 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 bivalent kit adopts fluorescent microspheres (202 nm) as an antibody marker, is divided into a 1 card and a 2 card which are integrated into a whole, and is respectively provided with a C line and a T line which are divided into a 1 card T1 line and a 2 card T2 line, and confirms that the novel coronavirus is positive by respectively diagnosing SARS-CoV NP protein and SARS-CoV protein negative.
Further, the SARS-CoV NP protein antibody marker is fluorescent microsphere (202 nm).
Further, the 1 card T1 line is coated with a monoclonal antibody of SARS-CoV NP.
Further, the 2-card T2 line is coated with a monoclonal antibody of SARS-CoV.
Further, the dual-card fluorescent microsphere is marked by a monoclonal antibody which can recognize SARS-CoV NP and SARS-CoV.
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 fluorescent quantitative rapid detection of new coronavirus (2019-nCoV) bivalent card.
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, SARS-CoV monoclonal antibody and SARS-CoV NP monoclonal antibody each 10-30ul, drawing line length C, T1 and T2, respectively, oven drying for use.
Marking SARS-CoV monoclonal antibody and SARS-CoV NP monoclonal antibody with 202nm fluorescent microsphere 10-50ul, packing into 96-hole non-adsorption enzyme label plate, and freeze drying. And (5) standby.
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, SARS-CoV monoclonal antibody and SARS-CoV NP monoclonal antibody each 10-30ul, drawing line length C, T1 and T2, respectively, oven drying for use.
Marking SARS-CoV monoclonal antibody and SARS-CoV NP monoclonal antibody with 202nm fluorescent microsphere 10-50ul, packing into 96-hole non-adsorption enzyme label plate, and freeze drying. And (5) standby.
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, respectively sucking 50ul to 2 freeze-dried fluorescent marker holes, blowing and beating uniformly, reacting for 5min, respectively sucking the liquid in the holes and dripping the liquid on sample pads of 1 card test strip and 2 card test strips, waiting for 10min, and then putting the sample 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 disclosed herein is a novel coronavirus (2019-nCoV) fluorescence quantitative rapid detection bivalent kit.
Claims (5)
1. A novel fluorescent quantitative rapid detection bigeminy kit for coronavirus is characterized in that: the bivalent kit adopts fluorescent microspheres (202 nm) as antibody markers, is divided into two unifications of 1 card and 2 cards, is respectively provided with a C line and a T line, and is divided into a 1 card T1 line and a 2 card T2 line, and can be used for determining the positivity of the novel coronavirus by respectively diagnosing the positivity of SARS-CoV NP protein and the negative of SARS-CoV protein.
2. The novel fluorescent quantitative rapid detection bivalent kit for coronavirus according to claim 1, which is characterized in that: the SARS-CoV NP protein antibody marker adopts fluorescent microspheres (202 nm).
3. The novel fluorescent quantitative rapid detection bivalent kit for coronavirus according to claim 1, which is characterized in that: the 1 card T1 line is coated with a monoclonal antibody of SARS-CoV NP.
4. The novel fluorescent quantitative rapid detection bivalent kit for coronavirus according to claim 1, which is characterized in that: the 2-card T2 coated monoclonal antibody of SARS-CoV.
5. The novel fluorescent quantitative rapid detection bivalent kit for coronavirus according to claim 1, which is characterized in that: the dual-card fluorescent microsphere is marked by a monoclonal antibody for identifying SARS-CoV NP and SARS-CoV.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010185501.7A CN111272722A (en) | 2020-03-17 | 2020-03-17 | Novel fluorescent quantitative rapid detection bigeminy card kit for coronavirus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010185501.7A CN111272722A (en) | 2020-03-17 | 2020-03-17 | Novel fluorescent quantitative rapid detection bigeminy card kit for coronavirus |
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| Publication Number | Publication Date |
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| CN111272722A true CN111272722A (en) | 2020-06-12 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202010185501.7A Pending CN111272722A (en) | 2020-03-17 | 2020-03-17 | Novel fluorescent quantitative rapid detection bigeminy card kit for coronavirus |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101825636A (en) * | 2010-05-19 | 2010-09-08 | 厦门大学附属中山医院 | Reagent strip for joint detection of syphilis specific IgM antibody and specific total antibody and preparation method thereof |
| 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 |
-
2020
- 2020-03-17 CN CN202010185501.7A patent/CN111272722A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101825636A (en) * | 2010-05-19 | 2010-09-08 | 厦门大学附属中山医院 | Reagent strip for joint detection of syphilis specific IgM antibody and specific total antibody and preparation method thereof |
| 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 |
Non-Patent Citations (3)
| Title |
|---|
| BO DIAO ET AL: "Human Kidney is a Target for Novel Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Infection", 《MEDRXIV》 * |
| 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》 * |
| 何长民等: "《生物制品基础》", 31 July 1986, 甘肃人民出版社 * |
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Application publication date: 20200612 |
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