CN111308078A - Colloidal gold bigeminy card for detecting novel coronavirus - Google Patents
Colloidal gold bigeminy card for detecting novel coronavirus Download PDFInfo
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- CN111308078A CN111308078A CN202010185500.2A CN202010185500A CN111308078A CN 111308078 A CN111308078 A CN 111308078A CN 202010185500 A CN202010185500 A CN 202010185500A CN 111308078 A CN111308078 A CN 111308078A
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- novel coronavirus
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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
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
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- 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
Abstract
The invention relates to a colloidal gold two-link card for detecting novel coronavirus. The test paper is characterized in that two cards are integrated, 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 present invention provides a colloidal gold two-joint card for detecting new type coronavirus, and said two-joint card can be used for respectively diagnosing SARS-CoV NP protein positive and SARS-CoV protein negative to define that the new type coronavirus is positive, and its application is convenient and quick, and is suitable for quickly diagnosing diseases resulted from new type coronavirus of various medical institutions.
The invention adopts the following technical scheme:
the colloidal gold duplex card for detecting the novel coronavirus is divided into 1 card and 2 cards which are combined into one card, the two cards are respectively provided with a C line and a T line, the T line is divided into a 1 card T1 line and a 2 card T2 line, and the colloidal gold duplex card is used for respectively diagnosing SARS-CoV NP protein positive and SARS-CoV protein negative to confirm the novel coronavirus positive.
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.
Furthermore, the colloidal gold test paper consists of a bottom plate, an nc membrane, absorbent paper, a gold label pad and a sample pad, wherein the dual-card gold label pad marks monoclonal antibodies for respectively identifying SARS-CoV NP and SARS-CoV.
The colloidal gold duplex card for detecting the novel coronavirus determines the positive of the novel coronavirus by respectively diagnosing the positive of SARS-CoV NP protein and the negative of SARS-CoV protein, is convenient and quick to use, and is suitable for quickly diagnosing 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 colloidal gold bivalent DNA of the novel coronavirus.
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 0.01M PB solution, adding 0.2% Tween 20 and 0.1% bovine serum albumin, soaking 300mm × 150mm gold label pad for 30min, and oven drying at 60 deg.C.
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, respectively marking C, T1 and T2, oven drying for use.
Marking SARS-CoV NP monoclonal antibody 10-50ul with four ten-thousandth chloroauric acid solution, spreading gold mark pad 5-10 strips for standby.
And assembling the colloidal gold test strip, namely sequentially assembling the base plate, the nc membrane, the absorbent paper, the gold label pad and the sample pad, and cutting the test strip.
The second embodiment is as follows:
preparing 0.01M tris solution, adding 0.2% Tween 20 and 0.1% bovine serum albumin, soaking 300mm × 150mm gold label pad for 30min, and oven drying at 60 deg.C for use.
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, respectively marking C, T1 and T2, oven drying for use.
Marking SARS-CoV NP monoclonal antibody 10-50ul with four ten-thousandth chloroauric acid solution, spreading gold mark pad
5-10 strips for standby.
And assembling the colloidal gold test strip, namely sequentially assembling the base plate, the nc membrane, the absorbent paper, the gold label pad and the sample pad, and cutting the test strip.
Application method of test paper
Diluting the quality control product or sample to 50 times, taking the test strip, respectively sucking 50ul of sample, dripping the sample on the sample pads of the card 1 and the card 2, waiting for 15min, and observing the outlet condition of the line c and the line t, 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 test paper is the colloidal gold binary card for detecting the novel coronavirus.
Claims (4)
1. A novel coronavirus colloidal gold two-link card is characterized in that: the colloidal gold bivalent card is divided into a 1 card and a 2 card which are integrated into a whole, the two cards are respectively provided with a C line and a T line, the T line is divided into a 1 card T1 line and a 2 card T2 line, and the colloidal gold bivalent card is used for respectively diagnosing SARS-CoV NP protein positivity and SARS-CoV protein negativity to determine novel coronavirus positivity.
2. The novel coronavirus detection colloidal gold test strip according to claim 1, wherein the test strip comprises: the 1 card T1 line is coated with a monoclonal antibody of SARS-CoV NP.
3. The novel coronavirus detection colloidal gold test strip according to claim 1, wherein the test strip comprises: the 2-card T2 coated monoclonal antibody of SARS-CoV.
4. The novel coronavirus colloidal gold-based two-part card according to claim 1, wherein: the colloidal gold duplex card consists of a bottom plate, an nc membrane, absorbent paper, a gold label pad and a sample pad, wherein the colloidal gold duplex card gold label pad is marked with monoclonal antibodies for respectively identifying SARS-CoV NP and SARS-CoV.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010185500.2A CN111308078A (en) | 2020-03-17 | 2020-03-17 | Colloidal gold bigeminy card for detecting novel coronavirus |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010185500.2A CN111308078A (en) | 2020-03-17 | 2020-03-17 | Colloidal gold bigeminy card for detecting novel coronavirus |
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| Publication Number | Publication Date |
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| CN111308078A true CN111308078A (en) | 2020-06-19 |
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| CN202010185500.2A Pending CN111308078A (en) | 2020-03-17 | 2020-03-17 | Colloidal gold bigeminy card for detecting novel coronavirus |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111518176A (en) * | 2020-07-06 | 2020-08-11 | 北京金智准科技有限公司 | Paired antigen for novel coronavirus antibody double-antigen sandwich detection, detection test paper and preparation method thereof |
Citations (5)
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| 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 |
| CN103983776A (en) * | 2014-05-27 | 2014-08-13 | 国家纳米科学中心 | Colloidal gold immunochromatography test strip for simultaneously detecting viral antigens and antibodies |
| CN105606801A (en) * | 2015-12-19 | 2016-05-25 | 中国科学院寒区旱区环境与工程研究所 | Lily X virus semi-quantitative detection gold-labeled card and preparation method thereof |
| CN108845149A (en) * | 2018-07-05 | 2018-11-20 | 郑州大学 | The colloidal gold duplex test strips and preparation method thereof of porcine reproductive and respiratory syndrome virus and swine fever virus are detected simultaneously |
| CN109596824A (en) * | 2019-01-04 | 2019-04-09 | 杭州奥泰生物技术股份有限公司 | A kind of Test paper and preparation method thereof of quick diagnosis Lyme disease |
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2020
- 2020-03-17 CN CN202010185500.2A patent/CN111308078A/en active Pending
Patent Citations (5)
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| 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 |
| CN103983776A (en) * | 2014-05-27 | 2014-08-13 | 国家纳米科学中心 | Colloidal gold immunochromatography test strip for simultaneously detecting viral antigens and antibodies |
| CN105606801A (en) * | 2015-12-19 | 2016-05-25 | 中国科学院寒区旱区环境与工程研究所 | Lily X virus semi-quantitative detection gold-labeled card and preparation method thereof |
| CN108845149A (en) * | 2018-07-05 | 2018-11-20 | 郑州大学 | The colloidal gold duplex test strips and preparation method thereof of porcine reproductive and respiratory syndrome virus and swine fever virus are detected simultaneously |
| CN109596824A (en) * | 2019-01-04 | 2019-04-09 | 杭州奥泰生物技术股份有限公司 | A kind of Test paper and preparation method thereof of quick diagnosis Lyme disease |
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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》 * |
| 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》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111518176A (en) * | 2020-07-06 | 2020-08-11 | 北京金智准科技有限公司 | Paired antigen for novel coronavirus antibody double-antigen sandwich detection, detection test paper and preparation method thereof |
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Application publication date: 20200619 |
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