CN113985028A - Diluent for colloidal gold method antigen detection - Google Patents
Diluent for colloidal gold method antigen detection Download PDFInfo
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- CN113985028A CN113985028A CN202111412878.2A CN202111412878A CN113985028A CN 113985028 A CN113985028 A CN 113985028A CN 202111412878 A CN202111412878 A CN 202111412878A CN 113985028 A CN113985028 A CN 113985028A
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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
- 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
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
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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
- G01N33/543—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
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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
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Abstract
The invention discloses a diluent for colloidal gold antigen detection, which is characterized in that: the method comprises the following raw materials: Tris-HCl buffer solution, NaCl, trehalose, Triton-X100, TCEP and BSA; wherein the Tris-HCl buffer solution has the molar concentration of 30-60mM, the molar concentration of NaCl is 100-200mM, the weight concentration of trehalose is 15-20%, the weight concentration of Triton-X100 is 0.1-3%, the weight concentration of TCEP is 0.1-3%, and the weight concentration of BSA is 0.05-3%. The immune colloidal gold test card diluent for the new coronavirus can promote the combination of virus antigen and colloidal gold, and has an important effect on antigen detection sensitivity. The diluent can improve the dilution multiple of the antigen and improve the sensitivity of antigen detection.
Description
Technical Field
The invention relates to the field of virus colloidal gold antigen detection, in particular to a diluent for detecting a new coronavirus by a colloidal gold antigen detection method.
Background
Coronaviruses are a large family of viruses that cause colds and more severe diseases such as Middle East Respiratory Syndrome (MERS) and Severe Acute Respiratory Syndrome (SARS). The novel coronavirus is a newly discovered new strain of coronavirus. At present, the detection method of the new coronavirus mainly comprises a nucleic acid method, an antigen and antibody detection method, wherein the nucleic acid detection method requires the steps of sampling by professional personnel, extracting and detecting nucleic acid in a laboratory and the like, and has the defects of easy exposure of workers, high risk, long time and the like. The antigen colloidal gold detection method is a method capable of quickly detecting the new coronavirus.
The immune colloidal gold technology is a novel immune marking technology which takes colloidal gold as a tracer marker or a color developing agent and is applied to antigen-antibody reaction. The use is convenient and quick, and the basic use and the field use are convenient; the cost is low, the technology does not need a special detection instrument, the application range is wide, and the technology can be suitable for various detection environments; it is widely used in the fields of immunology, histology, pathology and cell biology.
Disclosure of Invention
In order to make up for the defects of the prior art, the invention provides the antigen detection diluent for the colloidal gold method, which has sensitive reaction and convenient use.
The invention is realized by the following technical scheme:
the invention discloses a diluent for colloidal gold antigen detection, which is characterized in that: the method comprises the following raw materials:
Tris-HCl buffer solution, NaCl, trehalose, Triton-X100, TCEP and BSA;
wherein the Tris-HCl buffer solution has a Tris molar concentration of 30-60mM (millimole per liter),
the molar concentration of NaCl was 100-,
the weight concentration of the trehalose is 15-20%,
the weight concentration of Triton-X100 is 0.1-3%,
the weight concentration of TCEP is 0.1-3%,
the weight concentration of BSA is 0.05% -3%.
Furthermore, the diluent for detecting the antigen by the colloidal gold method also contains EDTA.2Na with the weight concentration of 0.1-3 percent and CA-630 with the weight concentration of 0.5-3 percent.
Further, the pH of the diluent for detecting the antigen by the colloidal gold method is adjusted to 7.0-8.0 by Hcl.
The invention relates to a diluent for colloidal gold antigen detection, which is prepared by the following steps: the raw materials are sequentially added into water and mixed together, water is added to prepare 1 liter of solution, and the solution is stirred uniformly and adjusted to the concentration.
NaCl provides inorganic salts, trehalose is a protectant, Triton-X100 (polyethylene glycol octylphenyl ether) is a surfactant, TCEP (trichloroethyl phosphate) is for interference removal, Ca-630 (nonionic surfactant Igepal CA-630) is a surfactant, and BSa (bovine serum albumin) is a protectant. Edta.2na (disodium edetate) is used as a chelating agent. Tris-HCl buffer (amino tri-butanol) was an aqueous solution.
The immune colloidal gold test card diluent for the new coronavirus can promote the combination of virus antigen and colloidal gold, and has an important effect on antigen detection sensitivity. The diluent can improve the dilution multiple of the antigen and improve the sensitivity of antigen detection.
Detailed Description
Example 1
The invention relates to a diluent for colloidal gold antigen detection, which comprises the following raw materials:
Tris-HCl buffer solution, NaCl, trehalose, Triton-X100, TCEP and BSA;
wherein Tris-HCl buffer solution has a Tris molar concentration of 30mM (millimole per liter), hydrochloric acid for adjusting the pH value to 7.0,
the weight concentration of EDTA.2Na is 0.1 percent,
the molar concentration of Nacl was 100mM (millimoles per liter),
the weight concentration of the trehalose is 15 percent,
the weight concentration of Triton-X100 is 0.1 percent,
the weight concentration of TCEP is 0.1%,
the weight concentration of CA-630 is 0.5%,
the weight concentration of BSA was 0.05%.
The invention relates to a diluent for colloidal gold antigen detection, which is prepared by the following steps: the raw materials are sequentially added into water and mixed together, water is added to prepare 1 liter of solution, and the solution is stirred uniformly and adjusted to the concentration.
Example 2
The invention relates to a diluent for colloidal gold antigen detection, which comprises the following raw materials:
Tris-HCl buffer solution, NaCl, trehalose, Triton-X100, TCEP and BSA;
wherein Tris-HCl buffer solution has a Tris molar concentration of 60mM (millimole per liter), hydrochloric acid for adjusting the pH value to 8.0,
the weight concentration of EDTA.2Na is 3 percent,
the molar concentration of Nacl was 200mM (millimoles per liter),
the weight concentration of the trehalose is 20 percent,
the weight concentration of Triton-X100 is 3 percent,
the TCEP concentration by weight was 3%,
the weight concentration of CA-630 is 3%,
the weight concentration of BSA was 3%.
The invention relates to a diluent for colloidal gold antigen detection, which is prepared by the following steps: the raw materials are sequentially added into water and mixed together, water is added to prepare 1 liter of solution, and the solution is stirred uniformly and adjusted to the concentration.
Example 3
The invention relates to a diluent for colloidal gold antigen detection, which comprises the following raw materials:
Tris-HCl buffer solution, NaCl, trehalose, Triton-X100, TCEP and BSA;
wherein Tris-HCl buffer solution has a Tris molar concentration of 40mM (millimole per liter), hydrochloric acid for adjusting the pH value to 7.5,
the weight concentration of EDTA.2Na is 2 percent,
the molar concentration of Nacl was 150mM (millimoles per liter),
the weight concentration of the trehalose is 17 percent,
the weight concentration of Triton-X100 is 2 percent,
the TCEP concentration by weight was 2%,
the weight concentration of CA-630 is 2%,
the weight concentration of BSA was 2%.
The invention relates to a diluent for colloidal gold antigen detection, which is prepared by the following steps: the raw materials are sequentially added into water and mixed together, water is added to prepare 1 liter of solution, and the solution is stirred uniformly and adjusted to the concentration.
Example 4
The invention relates to a diluent for colloidal gold antigen detection, which comprises the following raw materials:
Tris-HCl buffer solution, NaCl, trehalose, Triton-X100, TCEP and BSA;
wherein Tris-HCl buffer solution has a Tris molar concentration of 50mM (millimole per liter), hydrochloric acid for adjusting the pH value to 7.7,
the weight concentration of EDTA.2Na is 1 percent,
the molar concentration of Nacl was 180mM (millimoles per liter),
the weight concentration of the trehalose is 16 percent,
the weight concentration of Triton-X100 is 1 percent,
the TCEP concentration by weight was 1%,
the weight concentration of CA-630 is 1%,
the weight concentration of BSA was 1%.
The invention relates to a diluent for colloidal gold antigen detection, which is prepared by the following steps: the raw materials are sequentially added into water and mixed together, water is added to prepare 1 liter of solution, and the solution is stirred uniformly and adjusted to the concentration.
Example 5
The invention relates to a diluent for colloidal gold antigen detection, which comprises the following raw materials:
Tris-HCl buffer solution, NaCl, trehalose, Triton-X100, TCEP and BSA;
wherein Tris-HCl buffer solution has a Tris molar concentration of 50mM (millimole per liter), hydrochloric acid for adjusting the pH value to 7.8,
the molar concentration of Nacl was 150mM (millimoles per liter),
the weight concentration of the trehalose is 17 percent,
the weight concentration of Triton-X100 is 2 percent,
the TCEP concentration by weight was 2%,
the weight concentration of BSA was 2%.
The invention relates to a diluent for colloidal gold antigen detection, which is prepared by the following steps: the raw materials are sequentially added into water and mixed together, water is added to prepare 1 liter of solution, and the solution is stirred uniformly and adjusted to the concentration.
Example 6
The invention relates to a diluent for colloidal gold antigen detection, which comprises the following raw materials:
Tris-HCl buffer solution, NaCl, trehalose, Triton-X100, TCEP and BSA;
wherein the molar concentration of the Tris-HCl buffer solution is 40mM (millimole per liter), the PH value is adjusted to be 7.3 by hydrochloric acid,
the molarity of Nacl was 130mM (millimoles per liter),
the weight concentration of the trehalose is 16 percent,
the weight concentration of Triton-X100 is 1 percent,
the TCEP concentration by weight was 1%,
the weight concentration of BSA was 1%.
The invention relates to a diluent for colloidal gold antigen detection, which is prepared by the following steps: the raw materials are sequentially added into water and mixed together, water is added to prepare 1 liter of solution, and the solution is stirred uniformly and adjusted to the concentration.
Examples 7, 8: effect verification
(1) Preparing an antigen detection diluent A solution and an antigen detection diluent B solution, preparing according to the following components and concentrations, and adjusting the pH value to 7.5.
The preparation method is the same as that of example 1.
(2) The purchased new coronavirus N protein is diluted into different concentrations by A, B two diluents and dropped on a detection card for detection. If the detection card has C lines and T lines, the detection result shows that the N protein is detected to be positive, if only the C lines appear, the detection result shows that the N protein cannot be detected, if only the T lines appear, the detection result shows that the C lines do not appear, the result is invalid, and the detection is carried out again. The results are as follows:
note: + indicates the depth of color of the T-line, and the greater the + the darker the color, indicating a higher concentration of N protein. The C line appears each time.
From the result, the B diluent belongs to the conventional diluent, under the same condition, the N protein antigen cannot be detected to be positive after being diluted to a certain concentration, and the A diluent can be detected to be positive. The result shows that the dilution A can improve the minimum detection limit concentration and has higher sensitivity.
Claims (3)
1. A diluent for colloidal gold antigen detection is characterized in that: the method comprises the following raw materials:
Tris-HCl buffer solution, NaCl, trehalose, Triton-X100, TCEP and BSA;
wherein the molar concentration of the Tris-HCl buffer solution is 30-60mM,
the molar concentration of NaCl is 100-200mM,
the weight concentration of the trehalose is 15-20%,
the weight concentration of Triton-X100 is 0.1-3%,
the weight concentration of TCEP is 0.1-3%,
the weight concentration of BSA is 0.05% -3%.
2. The dilution for detecting antigen by colloidal gold method according to claim 1, which is characterized in that: also contains EDTA.2Na with weight concentration of 0.1-3%, and also contains CA-630 with weight concentration of 0.5-3%.
3. The dilution for detecting antigen by colloidal gold method according to claim 1, which is characterized in that: the pH value of the Tris-HCl buffer solution is adjusted to 7.0-8.0 by HCl.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202111412878.2A CN113985028A (en) | 2021-11-25 | 2021-11-25 | Diluent for colloidal gold method antigen detection |
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| CN202111412878.2A CN113985028A (en) | 2021-11-25 | 2021-11-25 | Diluent for colloidal gold method antigen detection |
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Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4931385A (en) * | 1985-06-24 | 1990-06-05 | Hygeia Sciences, Incorporated | Enzyme immunoassays and immunologic reagents |
| CN1675550A (en) * | 2002-08-01 | 2005-09-28 | Mtm实验室公司 | Method for solution based diagnosis |
| US20120295279A1 (en) * | 2010-02-05 | 2012-11-22 | Nichirei Biosciences Inc. | Pretreatment solution for immunohistochemical staining and condensed solution thereof |
| CN110954692A (en) * | 2019-12-20 | 2020-04-03 | 蓝怡科技集团股份有限公司 | Reducing agent buffer solution and preparation method and application thereof |
| CN111983217A (en) * | 2020-09-03 | 2020-11-24 | 菲鹏生物股份有限公司 | Sample treatment fluid and application thereof |
| CN112255421A (en) * | 2020-10-13 | 2021-01-22 | 黎法飓 | Lipoprotein a detection kit and detection method |
| CN112362869A (en) * | 2021-01-14 | 2021-02-12 | 山东康华生物医疗科技股份有限公司 | Multi-respiratory tract antigen detection card and kit |
| CN112557655A (en) * | 2021-01-06 | 2021-03-26 | 深圳容金科技有限公司 | Novel coronavirus, influenza A virus and influenza B virus antigen three-in-one detection kit and preparation and use methods thereof |
| CN112979810A (en) * | 2019-12-16 | 2021-06-18 | 广东菲鹏生物有限公司 | TAG-72 antibody-HRP (horse radish peroxidase) diluent and application thereof |
| WO2021202929A1 (en) * | 2020-04-02 | 2021-10-07 | Genalyte, Inc. | Rapid multiplexed serological test |
| WO2021205042A1 (en) * | 2020-04-10 | 2021-10-14 | RNAssist Limited | Composition and method for inactivating a virus |
| CN113533721A (en) * | 2021-07-15 | 2021-10-22 | 上海伯杰医疗科技有限公司北京分公司 | Colloidal gold method detection test strip for influenza A/B virus antigen and preparation method thereof |
-
2021
- 2021-11-25 CN CN202111412878.2A patent/CN113985028A/en active Pending
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4931385A (en) * | 1985-06-24 | 1990-06-05 | Hygeia Sciences, Incorporated | Enzyme immunoassays and immunologic reagents |
| CN1675550A (en) * | 2002-08-01 | 2005-09-28 | Mtm实验室公司 | Method for solution based diagnosis |
| US20120295279A1 (en) * | 2010-02-05 | 2012-11-22 | Nichirei Biosciences Inc. | Pretreatment solution for immunohistochemical staining and condensed solution thereof |
| CN112979810A (en) * | 2019-12-16 | 2021-06-18 | 广东菲鹏生物有限公司 | TAG-72 antibody-HRP (horse radish peroxidase) diluent and application thereof |
| CN110954692A (en) * | 2019-12-20 | 2020-04-03 | 蓝怡科技集团股份有限公司 | Reducing agent buffer solution and preparation method and application thereof |
| WO2021202929A1 (en) * | 2020-04-02 | 2021-10-07 | Genalyte, Inc. | Rapid multiplexed serological test |
| WO2021205042A1 (en) * | 2020-04-10 | 2021-10-14 | RNAssist Limited | Composition and method for inactivating a virus |
| CN111983217A (en) * | 2020-09-03 | 2020-11-24 | 菲鹏生物股份有限公司 | Sample treatment fluid and application thereof |
| CN112255421A (en) * | 2020-10-13 | 2021-01-22 | 黎法飓 | Lipoprotein a detection kit and detection method |
| CN112557655A (en) * | 2021-01-06 | 2021-03-26 | 深圳容金科技有限公司 | Novel coronavirus, influenza A virus and influenza B virus antigen three-in-one detection kit and preparation and use methods thereof |
| CN112362869A (en) * | 2021-01-14 | 2021-02-12 | 山东康华生物医疗科技股份有限公司 | Multi-respiratory tract antigen detection card and kit |
| CN113533721A (en) * | 2021-07-15 | 2021-10-22 | 上海伯杰医疗科技有限公司北京分公司 | Colloidal gold method detection test strip for influenza A/B virus antigen and preparation method thereof |
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