CN113820484A - Preparation method of household test strip for rapidly detecting dermatophytes - Google Patents
Preparation method of household test strip for rapidly detecting dermatophytes Download PDFInfo
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- CN113820484A CN113820484A CN202110992364.2A CN202110992364A CN113820484A CN 113820484 A CN113820484 A CN 113820484A CN 202110992364 A CN202110992364 A CN 202110992364A CN 113820484 A CN113820484 A CN 113820484A
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- dermatophyte
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- 241001480043 Arthrodermataceae Species 0.000 title claims abstract description 44
- 230000037304 dermatophytes Effects 0.000 title claims abstract description 44
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 238000012360 testing method Methods 0.000 title claims abstract description 10
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 24
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 16
- 239000000427 antigen Substances 0.000 claims abstract description 9
- 102000036639 antigens Human genes 0.000 claims abstract description 9
- 108091007433 antigens Proteins 0.000 claims abstract description 9
- 238000006243 chemical reaction Methods 0.000 claims abstract description 8
- 239000012528 membrane Substances 0.000 claims description 15
- 239000002245 particle Substances 0.000 claims description 9
- 238000001514 detection method Methods 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 239000001963 growth medium Substances 0.000 claims description 5
- 239000002250 absorbent Substances 0.000 claims description 3
- 230000002745 absorbent Effects 0.000 claims description 3
- 238000010521 absorption reaction Methods 0.000 claims description 3
- 238000004140 cleaning Methods 0.000 claims description 3
- 238000005238 degreasing Methods 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 238000000227 grinding Methods 0.000 claims description 3
- 230000036046 immunoreaction Effects 0.000 claims description 3
- 239000012535 impurity Substances 0.000 claims description 3
- 230000000415 inactivating effect Effects 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 238000003908 quality control method Methods 0.000 claims description 3
- 238000012795 verification Methods 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 241000223238 Trichophyton Species 0.000 claims 1
- 206010012504 Dermatophytosis Diseases 0.000 abstract description 4
- 241001460074 Microsporum distortum Species 0.000 abstract description 4
- 208000002474 Tinea Diseases 0.000 abstract description 4
- 239000003814 drug Substances 0.000 abstract description 3
- 206010017533 Fungal infection Diseases 0.000 abstract description 2
- 208000031888 Mycoses Diseases 0.000 abstract description 2
- 208000010195 Onychomycosis Diseases 0.000 abstract description 2
- 230000000843 anti-fungal effect Effects 0.000 abstract description 2
- 229940121375 antifungal agent Drugs 0.000 abstract description 2
- 229940079593 drug Drugs 0.000 abstract description 2
- 201000005882 tinea unguium Diseases 0.000 abstract description 2
- 238000003745 diagnosis Methods 0.000 abstract 1
- 208000015181 infectious disease Diseases 0.000 abstract 1
- 201000004624 Dermatitis Diseases 0.000 description 2
- 241000223229 Trichophyton rubrum Species 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- 208000022535 Infectious Skin disease Diseases 0.000 description 1
- 201000004681 Psoriasis Diseases 0.000 description 1
- 206010047601 Vitamin B1 deficiency Diseases 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 208000002894 beriberi Diseases 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000004092 self-diagnosis Methods 0.000 description 1
- 208000017520 skin disease Diseases 0.000 description 1
Classifications
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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
- G01N33/56961—Plant cells or fungi
-
- 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
-
- 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
-
- 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
- G01N2333/37—Assays involving biological materials from specific organisms or of a specific nature from fungi
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- General Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Pathology (AREA)
- Botany (AREA)
- Mycology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a preparation method of a household test strip for rapidly detecting dermatophyte, which mainly comprises the following technical key points: obtaining dermatophyte antigens; preparing dermatophyte monoclonal antibody; preparing a colloidal gold reagent strip for antigen-antibody reaction; preparing a clinical sample treatment solution; and preparing a complete reagent strip set and an instruction for use. The invention adopts the mode of the colloidal gold reagent strip to rapidly detect dermatophytes, is suitable for household or basic unit use, self-checks whether dermatophytes infection such as dermatophytosis or onychomycosis is suffered, and then goes to an internet hospital for inquiry or goes to a drugstore or purchases antifungal medicines on the internet, thereby effectively reducing the occurrence of misdiagnosis and mistreatment of superficial fungal infection and the economic and time cost of visiting a hospital for diagnosis.
Description
Technical Field
The invention relates to a preparation method of a household test strip for rapidly detecting dermatophyte, belonging to the technical field of biological medicine.
Background
Dermatophytosis is a high-incidence infectious skin disease, can occur in healthy people and people with immune insufficiency, and has high recurrence rate; since the clinical manifestations of dermatophytosis are sometimes similar to those of other skin diseases such as dermatitis, psoriasis, eczema, etc., the detection of persons with immune insufficiency in particular is more difficult; the existing dermatophyte detection mode is complex and is not suitable for home use, so that the personal detection cost is high, and time and labor are wasted.
Disclosure of Invention
In view of the above technical problems, the present invention aims to: provides a preparation method of a household test strip for rapidly detecting dermatophyte.
The technical solution of the invention is realized as follows: a preparation method of a household test strip for rapidly detecting dermatophyte comprises the following steps:
firstly, obtaining dermatophyte antigens;
② preparing dermatophyte monoclonal antibody;
preparing a colloidal gold reagent strip for antigen-antibody reaction;
preparing a clinical sample treatment solution;
preparing complete reagent strip set and instruction.
Preferably, in the step (i), the process for obtaining dermatophyte antigens comprises the following steps:
inoculating dermatophyte into a culture medium, cleaning and inactivating the dermatophyte in an aseptic environment after the dermatophyte grows up, drying the dermatophyte, grinding the dermatophyte into powder, extracting by using an alkaline extracting solution after degreasing, filtering by using filter paper, detecting and correcting a pH value, performing aseptic filtration and verification, inoculating the dermatophyte into the culture medium for culture, and finally obtaining the dermatophyte antigen.
Preferably, in the third step, the colloidal gold reagent strip comprises a supporting pad, and a sample pad, a colloidal gold pad, an NC membrane and a water absorption pad which are arranged on the supporting pad;
the sample pad is used for slowing down the sample permeation speed, removing sample impurities and adjusting the pH value and viscosity of the sample;
the colloidal gold pad is used for adsorbing gold-labeled conjugate particles, adsorbing and continuously transferring a sample to an NC membrane, maintaining the stability of the gold-labeled conjugate particles and ensuring the quantitative and complete release of the gold-labeled conjugate particles;
the NC membrane is provided with a detection line and a quality control line, a sample flows on the NC membrane and is mixed with a reagent to generate immunoreaction, and the reaction is developed on the NC membrane;
the absorbent pad is used to control the flow rate of the sample and promote the siphoning action.
Due to the application of the technical scheme, compared with the prior art, the invention has the following advantages:
the invention adopts the mode of the colloidal gold reagent strip to rapidly detect dermatophytosis, is suitable for household or basic unit use, self-checks whether beriberi or onychomycosis is suffered, then automatically goes to a drugstore or purchases antifungal medicines on the internet, and can effectively reduce misdiagnosis and mistreatment of superficial fungal infection.
Detailed Description
The present invention will be described with reference to examples.
The invention discloses a preparation method of a household test strip for rapidly detecting dermatophyte, which comprises the following steps:
firstly, obtaining general and specific antigens of dermatophytes; inoculating dermatophyte (Trichophyton rubrum) into a culture medium, cleaning and inactivating the dermatophyte in an aseptic environment after the dermatophyte grows up, drying the dermatophyte, grinding the dermatophyte into powder, extracting with an alkaline extract after degreasing, filtering with filter paper, detecting and correcting a pH value, performing aseptic filtration and verification, inoculating the dermatophyte in the culture medium, and culturing to obtain the dermatophyte antigen.
② preparing dermatophyte monoclonal antibody;
the research literature is attached with a standard monoclonal antibody preparation process, and after the preparation of the monoclonal antibody, the monoclonal antibody is subjected to antigen-antibody reaction with other common dermatophytes except trichophyton rubrum to verify the genus specificity of the monoclonal antibody.
Preparing a colloidal gold reagent strip for antigen-antibody reaction;
the colloidal gold reagent strip comprises a supporting pad, and a sample pad, a colloidal gold pad, an NC membrane and a water absorption pad which are arranged on the supporting pad;
the sample pad is used for slowing down the sample permeation speed, removing sample impurities and adjusting the pH value and viscosity of the sample;
the colloidal gold pad is used for adsorbing gold-labeled conjugate particles, adsorbing and continuously transferring a sample to an NC membrane, maintaining the stability of the gold-labeled conjugate particles and ensuring the quantitative and complete release of the gold-labeled conjugate particles;
the NC membrane is provided with a detection line and a quality control line, a sample flows on the NC membrane and is mixed with a reagent to generate immunoreaction, and the reaction is developed on the NC membrane;
the absorbent pad is used to control the flow rate of the sample and promote the siphoning action.
Preparing a clinical sample treatment solution;
preparing complete reagent strip set and instruction, packing into box and packing to obtain the product suitable for household self diagnosis and detection.
The above-mentioned embodiments are merely illustrative of the technical idea and features of the present invention, and the purpose thereof is to enable those skilled in the art to understand the contents of the present invention and implement the present invention, and not to limit the scope of the present invention, and all equivalent changes or modifications made according to the spirit of the present invention should be covered in the scope of the present invention.
Claims (3)
1. A preparation method of a household test strip for rapidly detecting dermatophyte is characterized by comprising the following steps:
firstly, obtaining dermatophyte antigens;
② preparing dermatophyte monoclonal antibody;
preparing a colloidal gold reagent strip for antigen-antibody reaction;
preparing a clinical sample treatment solution;
preparing complete reagent strip set and instruction.
2. The method for preparing a household test strip for rapidly detecting dermatophytes according to claim 1, wherein the method comprises the following steps:
in the step I, the process for obtaining the dermatophyte antigen is as follows:
inoculating dermatophyte into a constant-temperature incubator at 27 ℃ for culture, cleaning and inactivating a dermatophyte colony in an aseptic environment after the dermatophyte colony grows up, drying the dermatophyte, grinding the dermatophyte into powder, extracting with an alkaline extract after degreasing, filtering with filter paper, detecting and correcting a pH value, performing aseptic filtration and verification, inoculating the trichophyton in the culture medium for culture, and finally obtaining the dermatophyte antigen.
3. The method for preparing a household test strip for rapidly detecting dermatophytes according to claim 1, wherein the method comprises the following steps:
in the third step, the colloidal gold reagent strip comprises a supporting pad, and a sample pad, a colloidal gold pad, an NC membrane and a water absorption pad which are arranged on the supporting pad;
the sample pad is used for slowing down the sample permeation speed, removing sample impurities and adjusting the pH value and viscosity of the sample;
the colloidal gold pad is used for adsorbing gold-labeled conjugate particles, adsorbing and continuously transferring a sample to an NC membrane, maintaining the stability of the gold-labeled conjugate particles and ensuring the quantitative and complete release of the gold-labeled conjugate particles;
the NC membrane is provided with a detection line and a quality control line, a sample flows on the NC membrane and is mixed with a reagent to generate immunoreaction, and the reaction is developed on the NC membrane;
the absorbent pad is used to control the flow rate of the sample and promote the siphoning action.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110992364.2A CN113820484A (en) | 2021-08-27 | 2021-08-27 | Preparation method of household test strip for rapidly detecting dermatophytes |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110992364.2A CN113820484A (en) | 2021-08-27 | 2021-08-27 | Preparation method of household test strip for rapidly detecting dermatophytes |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113820484A true CN113820484A (en) | 2021-12-21 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110992364.2A Pending CN113820484A (en) | 2021-08-27 | 2021-08-27 | Preparation method of household test strip for rapidly detecting dermatophytes |
Country Status (1)
| Country | Link |
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| CN (1) | CN113820484A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116284366A (en) * | 2023-05-25 | 2023-06-23 | 南京黎明生物制品有限公司 | Quick detection antibody and kit for dermatophytosis and preparation method thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004159593A (en) * | 2002-11-14 | 2004-06-10 | Toyobo Co Ltd | Monoclonal antibody against dermatophyte, hybridoma producing the antibody and method for producing the antibody |
| JP2004258024A (en) * | 2003-02-03 | 2004-09-16 | Toyobo Co Ltd | Dermatophyte detection method, reagent for dermatophyte detection, and antigenicity activation method |
| CN106279349A (en) * | 2016-08-31 | 2017-01-04 | 广东工业大学 | For preparing extracting method and the preparation method of yolk antibody thereof of the dermatophytosis suppressor proteins antigen of yolk antibody |
| CN108690881A (en) * | 2017-03-30 | 2018-10-23 | 欧蒙医学诊断技术有限公司 | Measuring method for diagnosing dermatophytosis |
| CN112014565A (en) * | 2020-10-31 | 2020-12-01 | 南京黎明生物制品有限公司 | Test strip and kit for detecting trichophyton rubrum and preparation method of test strip |
-
2021
- 2021-08-27 CN CN202110992364.2A patent/CN113820484A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004159593A (en) * | 2002-11-14 | 2004-06-10 | Toyobo Co Ltd | Monoclonal antibody against dermatophyte, hybridoma producing the antibody and method for producing the antibody |
| JP2004258024A (en) * | 2003-02-03 | 2004-09-16 | Toyobo Co Ltd | Dermatophyte detection method, reagent for dermatophyte detection, and antigenicity activation method |
| CN106279349A (en) * | 2016-08-31 | 2017-01-04 | 广东工业大学 | For preparing extracting method and the preparation method of yolk antibody thereof of the dermatophytosis suppressor proteins antigen of yolk antibody |
| CN108690881A (en) * | 2017-03-30 | 2018-10-23 | 欧蒙医学诊断技术有限公司 | Measuring method for diagnosing dermatophytosis |
| CN112014565A (en) * | 2020-10-31 | 2020-12-01 | 南京黎明生物制品有限公司 | Test strip and kit for detecting trichophyton rubrum and preparation method of test strip |
Non-Patent Citations (2)
| Title |
|---|
| Y. TSUNEMI等: "Screening for tinea unguium by Dermatophyte Test Strip", 《BRITISH JOURNAL OF DERMATOLOGY》 * |
| 李东霞 等: "皮肤癣菌抗原制备的研究及其临床应用", 《中国皮肤性病学杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116284366A (en) * | 2023-05-25 | 2023-06-23 | 南京黎明生物制品有限公司 | Quick detection antibody and kit for dermatophytosis and preparation method thereof |
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Application publication date: 20211221 |