CN113317572A - Visual rapid detection mask for exhaled coronavirus and preparation method thereof - Google Patents
Visual rapid detection mask for exhaled coronavirus and preparation method thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A41—WEARING APPAREL
- A41D—OUTERWEAR; PROTECTIVE GARMENTS; ACCESSORIES
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- 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
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- 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
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Abstract
The invention relates to a visual rapid detection mask for exhaled coronavirus, which comprises a mask main body, a breather valve and a test strip, wherein the breather valve is arranged on the mask main body; the test strip comprises a filter disc, and a sample pad, a combination pad, a color development pad and a water absorption pad which are sequentially overlapped; the binding pad comprises colloidal gold coated with coronavirus N protein antibody; the color development pad is provided with a detection line and a quality control line, the detection line is coated with a coronavirus E protein antibody, and the quality control line is coated with an anti-coronavirus N protein antibody IgG. The test subject can realize the rapid and accurate detection of the coronavirus within 0.5-1h by wearing the mask. The mask takes the exhaled air as a detection object, and the exhaled air carries larger virus amount relative to body fluids such as sputum and saliva, so that suspected patients who cannot be diagnosed temporarily and other people (susceptible people, old people, children and the like) who are inconvenient to go to a hospital for detection can be rapidly preliminarily diagnosed by using the mask.
Description
Technical Field
The invention relates to the technical field of medical supplies, in particular to a visual rapid detection mask for exhaled breath coronavirus and a preparation method thereof.
Background
Coronaviruses are a group of pathogens that cause mainly respiratory and intestinal diseases. These viruses have many protrusions on their surface, like a crown of royal hat, and are therefore named coronaviruses. The coronavirus is a positive strand single strand RNA virus with an outer mantle (envelope), the diameter of the coronavirus is about 80-120 nm, and the genetic material of the coronavirus is the largest of all RNA viruses and infects human, mice, pigs, cats, dogs, wolfs, chickens, cows and poultry vertebrates. A variant of coronavirus, SARS-CoV, is a causative agent of Severe Acute Respiratory Syndrome (SARS) and belongs to the RNA virus.
The coronavirus was originally isolated from chicken in 1937, and the virus particles had a diameter of 60-200 nm, an average diameter of 100nm, a spherical or elliptical shape, and a polymorphism. Coronaviruses have envelopes, spinous processes exist on the envelopes, the whole virus is like coronas, and the spinous processes of different coronaviruses have obvious differences. Tubular inclusions are sometimes visible when the coronavirus infects the cell. To date, 7 types of coronaviruses known to infect humans are: 2019 the novel coronavirus (2019-nCoV) is the 7 th coronavirus which is known to infect human, and the other 6 are HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV and MERS-CoV, respectively. After people are infected with coronavirus, the common signs of the person are respiratory symptoms, fever, cough, shortness of breath, dyspnea and the like. In more severe cases, the infection can lead to pneumonia, severe acute respiratory syndrome, renal failure, and even death. There is currently no specific treatment for 2019 new coronavirus-induced diseases.
Currently, the detection methods for 2019 novel coronaviruses mainly include chest CT, reverse transcription polymerase chain reaction (RT-PCR), and the like, but these methods require the help of professionals and professional equipment, and can only be generally detected in hospitals or professional institutions, which results in the application of these detection methods being greatly limited. Secondly, a large number of suspected patients enter a hospital for detection, which greatly increases the risk of cross-infection and also leads to a shortage of medical resources. In order to deal with sudden epidemic situations, a rapid diagnosis method is urgently needed for home screening and preliminary screening of suspected patients, so that the suspected patients which cannot be diagnosed for a while and other people (susceptible people, old people, children and the like) which cannot go to hospital for detection can be diagnosed preliminarily and rapidly.
Disclosure of Invention
The invention aims to provide a visual rapid detection mask for exhaled breath coronavirus and a preparation method thereof, wherein the mask can be used for conveniently and rapidly detecting whether the exhaled breath contains the coronavirus, is a supplement to the existing coronavirus detection method, and can be used for remarkably improving the diagnosis efficiency and the detection accuracy particularly for 2019 novel coronavirus.
To this end, the invention provides a visual rapid detection mask for the exhaled coronary viruses, which comprises a mask body, a breather valve and a test strip;
the test strip is arranged on the breather valve and used for receiving the exhaled breath to be tested;
the test strip comprises a filter disc, a sample pad, a combination pad, a color development pad and a water absorption pad;
the sample pad, the combination pad, the color development pad and the water absorption pad are sequentially overlapped and are arranged on the filter sheet in a fitting manner;
the sample pad is positioned on the upper surface of the filter disc and is provided with a water absorption coating; the upper surface of the filter sheet faces the inner side of the mask main body;
the binding pad comprises colloidal gold coated with coronavirus N protein antibody;
the color development pad is positioned on the lower surface of the filter disc and is provided with a detection line and a quality control line, the detection line is close to one side of the combination pad, and the quality control line is close to one side of the water absorption pad; the detection line is coated with a coronavirus E protein antibody, and the quality control line is coated with an anti-coronavirus N protein antibody IgG;
the water absorption pad is positioned on the lower surface of the filter disc.
When a testee wears the visual rapid detection mask for breathing, the breathing valve is a one-way air outlet valve, the exhaled air is mainly discharged through the breathing valve, the sample pad of the test strip arranged on the breathing valve receives a large amount of exhaled air, and under the adsorption action of the water absorption pad, water vapor in the exhaled air sequentially passes through the sample pad, the combination pad and the color development pad and reaches the water absorption pad. In the process, coronavirus (if any) in the exhaled breath is specifically combined with a corresponding antibody, and chromogenic detection is realized according to the immune reaction principle.
Further, the mask body is selected from a disposable medical mask, a medical surgical mask, an industrial protective mask or a civil protective mask. In a specific embodiment, an N95 mask, such as a 3M9501V mask, is selected.
Further, the material of the water absorption coating of the sample pad comprises one or more of MOFs-HKUST-1, Cr-soc-MOF-1, NU-1500-Cr, activated carbon and porous silicon.
Further, the particle size of the colloidal gold is 10-50 nm.
Further, the coronavirus N protein antibody comprises rabbit anti-SARS Nsp3 antibody and rabbit MERS & SARS-CoV NP antibody.
Further, the binding concentration of the coronavirus N protein antibody coated colloidal gold is 30-60 mu g/ml.
Further, the coronavirus E protein antibody comprises a rabbit anti-SARS E protein antibody.
Further, the coating concentration of the coronavirus E protein antibody is 1-5 mu g/cm.
Furthermore, the anti-coronavirus E protein antibody IgG is a goat anti-rabbit IgG monoclonal antibody.
Further, the coating concentration of the anti-coronavirus E protein antibody IgG is 1-5 mu g/cm.
The second aspect of the present invention provides a preparation method of the mask for visual rapid detection of exhaled breath coronavirus, comprising:
(1) coating the surface of the sample pad with the water-absorbing coating, wherein the thickness is 1-2 mm;
(2) coating the coronavirus N protein antibody on the colloidal gold, uniformly coating the colloidal gold coated with the coronavirus N protein antibody on the surface of the binding pad, and airing at room temperature;
(3) respectively engraving a detection line and a quality control line on the developing pad, coating a coronavirus E protein antibody on the detection line, coating an anti-coronavirus N protein antibody IgG on the quality control line, and drying at room temperature;
(4) sequentially adhering the sample pad prepared in the step (1), the bonding pad prepared in the step (2), the color development pad prepared in the step (3) and the water absorption pad on a filter sheet to prepare the test strip;
(5) taking the mask body provided with the breather valve, fixing the test strip prepared in the step (4) on the breather valve, and making the sample pad face the inner side of the mask, namely preparing the visual rapid detection mask for the exhaled coronary artery disease.
In a specific embodiment, the material of the water-absorbing coating is MOFs-HKUST-1, and the step (1) comprises the following steps: mixing a copper nitrate solution and an ethanolamine solution, standing at room temperature for 24-36h, and performing suction filtration through an organic phase filter membrane to obtain a first film material; then reacting the first film material with an ethanol water solution of trimesic acid at normal temperature for 0.5-2h to obtain a second film material; a second thin film material is applied to the surface of the sample pad.
Further, in the step (2), the preparation method of the colloidal gold comprises: taking chloroauric acid as a main raw material, and preparing the colloidal gold through a reduction reaction; the reducing agent is one or more selected from sodium citrate, tannic acid, ascorbic acid, white phosphorus and sodium borohydride; sodium citrate is preferred.
A third aspect of the present invention provides a method for detecting an exhaled breath coronavirus, comprising the steps of:
the visual rapid detection mask is worn by a subject for 0.5 to 1 hour; and observing the color change of the detection line and the quality control line.
The result determination method is as follows:
red strips appear on the quality control line and the detection line, and the result is judged to be positive;
only a red strip appears on the quality control line, and the result is judged to be negative;
and the quality control line has no red strip, and the result is judged to be invalid no matter whether the detection line has the red strip or not.
The invention takes the exhaled breath of the subject as the starting point, compared with sputum and saliva, the exhaled breath has large carrying amount and high accuracy of the exhaled breath virus, and the diagnosis rate can be obviously enhanced particularly for suspected patients in early stage or latent stage. In order to enhance the sensitivity and convenience of exhaled breath detection, the invention designs a brand-new visual detection test strip which can be directly inlaid at a mask air valve to achieve the functions of capturing and enriching water and viruses in real time. The visual rapid detection mask and the corresponding detection method provided by the invention are important supplements to the existing detection means, greatly improve the rapid detection of the novel coronavirus, and particularly can obviously enhance the diagnosis rate for early or latent suspected patients.
Compared with the prior art, the invention has the following advantages:
(1) the visual rapid detection mask provided by the invention integrates a nano MOF coating preparation technology, a monoclonal antibody preparation technology, a nano gold labeling technology, an immunodetection technology and a membrane chromatography technology; the water absorption coating of the sample pad is utilized to efficiently adsorb and capture water vapor and viruses in the exhaled breath; the characteristic of highly specific combination of antigen-antibody reaction is utilized to efficiently, specifically, sensitively and rapidly identify viruses; and the qualitative detection result can be obtained by visual inspection through an immune gold labeling technology without the help of professional instruments and equipment.
(2) The invention takes the expired air of the subject as a detection object, and compared with body fluids such as sputum, saliva and the like, the virus carrying capacity of the expired air is larger, so that the visual rapid detection mask provided by the invention can realize accurate diagnosis on early or latent suspected patients.
(3) Compared with the detection methods such as the current commonly used electron microscope technology, enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR) and the like, the visual rapid detection mask provided by the invention is simple to operate, convenient to carry and low in cost, can realize rapid detection only by wearing for 0.5h, and is suitable for home screening.
(4) When an Anti-SARS E protein antibody, an Anti-SARS N protein antibody and an Anti-SARS N protein antibody goat IgG polyclonal antibody are specifically selected, the visual rapid detection mask provided by the invention can rapidly diagnose whether a subject is infected with 2019 novel coronavirus or SARS virus, and particularly can significantly enhance the diagnosis rate for early or latent suspected patients.
Drawings
Various other advantages and benefits will become apparent to those of ordinary skill in the art upon reading the following detailed description of the preferred embodiments. The drawings are only for purposes of illustrating the preferred embodiments and are not to be construed as limiting the invention. Also, like reference numerals are used to refer to like parts throughout the drawings. In the drawings:
FIG. 1 is a schematic view of a visual rapid-detection mask for exhaled breath coronaviruses of the present invention;
FIG. 2 is a schematic front view of a test strip;
FIG. 3 is a schematic reverse side view of the test strip;
FIG. 4 is an electron micrograph representation of the MOF coated upper surface of the sample pad;
FIG. 5 is an electron micrograph of the MOF coated side of the sample pad;
FIG. 6 is a comparison between before and after coating of a conjugate pad with colloidal gold, FIG. 6A shows a photograph of a conjugate pad without coated with colloidal gold, and FIG. 6B shows a photograph of a conjugate pad coated with colloidal gold coated with coronavirus N-protein antibody;
1-mask body, 2-breather valve, 3-test paper strip, 31-filter disc, 32-sample pad, 33-combination pad, 34-color pad, 341-detection line, 342-quality control line and 35-water absorption pad.
Detailed Description
Exemplary embodiments of the present disclosure will be described in more detail below with reference to the accompanying drawings. While exemplary embodiments of the present disclosure are shown in the drawings, it should be understood that the present disclosure may be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.
Example 1
The embodiment provides a visual rapid detection mask for an exhaled breath coronavirus, and referring to fig. 1 and 2, the visual rapid detection mask comprises a mask main body 1, a breather valve 2 and a test strip 3, wherein the test strip 3 is arranged on the breather valve 2 and is used for receiving an exhaled breath to be detected; the test strip 3 comprises a filter disc 31, a sample pad 32, a combination pad 33, a color development pad 34 and a water absorption pad 35; the sample pad 32, the combination pad 33, the color development pad 34 and the water absorption pad 35 are sequentially overlapped and are arranged on the filter disc 31 in a fitting manner; the sample pad 32 is located on the upper surface of the filter 31 and has a water-absorbing coating; the upper surface of the filter 31 faces the inner side of the mask body 1; the conjugate pad 33 comprises colloidal gold coated with coronavirus N protein antibody; the color development pad 34 is positioned on the lower surface of the filter disc 31 and is provided with a detection line 341 and a quality control line 342, the detection line 341 is close to one side of the combination pad 33, and the quality control line 342 is close to one side of the water absorption pad 35; the detection line 341 is coated with an antibody of coronavirus E protein, and the quality control line 342 is coated with an antibody IgG of anti-coronavirus N protein; the absorbent pad 35 is located on the lower surface of the filter 31.
When the testee wears the visual quick detection gauze mask is breathed, because breather valve 2 is the one-way air outlet valve, the expiratory gas is mainly discharged through breather valve 2, locates the sample pad 32 of the test paper strip 3 on breather valve 2 and receives a large amount of expiratory gases, and under the adsorption of water absorption pad 35, the steam in the expiratory gas passes through sample pad 32, combination pad 33, colour development pad 34 in proper order, reaches water absorption pad 35. In the process, coronavirus (if any) in the exhaled breath is specifically combined with a corresponding antibody, and chromogenic detection is realized according to the immune reaction principle.
Example 2
This embodiment has prepared a visual quick detection gauze mask for exhaled breath coronavirus, includes:
1 materials and reagents
1.1 materials
The 3M9501V mask was purchased from beijing yutong hong ye scientific and technological development ltd. The filter, sample pad, absorbent paper, conjugate pad, color pad, and absorbent pad were purchased from Beijing Aidekang scientific Co.
1.2 antibodies
Anti-SARS E antibody, Anti-SARS N protein antibody, Anti-SARS N protein antibody goat IgG polyclonal antibody was purchased from Abcam, UK.
2 method of experiment
2.1 preparation of absorbent MOF sample pad
The same volume of copper nitrate solution (4mM) and ethanolamine solution were mixed rapidly and allowed to stand at room temperature for 36 h. 60mL of the reaction solution was filtered through a nylon 66 organic phase filter (0.22 μm) to obtain a pale blue thin film material. The resulting film material was then reacted with 15mL of trimesic acid (5mM) in aqueous ethanol (v: v ═ 1:1) at room temperature for 1h to give a dark blue HKUST-1 film. A cellulose membrane strip was selected as a sample pad, sprayed with PBS buffer containing 5% BSA, 0.1% Tween 20, and air-dried at room temperature. And coating the MOFs-HKUST-1 film on the surface of the sample pad, standing and air-drying at room temperature.
2.2 preparation of conjugate pads
Selecting a glass cellulose band as a solid phase material of the binding pad, spraying colloidal gold coated with an N protein antibody on the band, and air-drying at room temperature.
2.3 preparation of color pads
Drawing a quality control line and a detection line on the color pad, wherein the distance between the quality control line and the detection line is 0.5-0.8cm, respectively coating anti-SARS N protein antibody goat IgG and E protein antibody on the quality control line and the detection line, and air-drying at room temperature.
2.4 assembling and detecting mask
The sample pad, the combination pad, the color development pad, the water absorption pad and the filter disc are installed well as shown in figure 2, then the sample pad, the combination pad, the color development pad, the water absorption pad and the filter disc are cut into test strips by a cutting machine, the test strips are embedded into the air valve of the respirator, one surface of the sample pad faces the inner side of the respirator, and the visual rapid detection respirator for the exhaled coronavirus is prepared. The masks prepared in this example were used in examples 3 to 6.
Example 3 qualitative assay
Blowing the sample pad with the expired air of the patient infected with 2019 novel coronavirus for 0.5h under aseptic condition, standing for 5min, and observing color changes of the detection line and the quality control line; the comparison was performed with the stock solution of the gasified 2019 novel coronavirus as a control group.
The experimental results show that the quality control lines of the gasified coronavirus stock solution group and the patient exhaled breath group are red, which indicates that the data are valid. Red bands are found on the detection lines, which shows that the reagent strip provided by the invention can be used for qualitatively detecting 2019 novel coronaviruses.
Example 4 specific assay
The visual rapid detection mask prepared in example 1 was worn by healthy persons, influenza patients and new coronary pneumonia patients (infected with 2019 novel coronavirus), respectively, and the wearing time was 0.5 h. And observing the color change of the detection line and the quality control line.
The experimental results show that the quality control lines of the three groups of samples are all red, which indicates that the data is valid. The detection lines of the healthy people group and the influenza patient group have no color change, which indicates that the new coronavirus infection of 2019 does not exist. In the new coronary pneumonia patient group, the detection line is obviously red, which indicates that the patients in the group are infected with 2019 novel coronavirus.
Example 5 sensitivity measurement
Setting the concentration gradient to 10 in sequence-15M、10-14M、10-13M、10-12M、10-11M and 10-10M's 2019 novel coronavirus solution, with solution gasification simulation patient's exhale, blow exhale on the test paper strip of visual short-term test gauze mask pneumatic valve, the time is 0.5h, observes under the different concentration conditions, the colour change of chromogenic pad detection line. The results show that when the solution concentration is 10 or more-14When M is higher than M, the color change can be observed on the detection line, which indicates that the detection interval of the mask is more than or equal to 10-14M。
Example 6 clinical patient testing
Since the virus content in the exhaled breath varies per unit time for patients with different degrees of infection, the present example determines the detection time required for patients with different degrees of infection based on the detection results of clinical patients.
Patients infected with coronavirus were set as early stage, middle stage and late stage patient groups, respectively, according to the severity of infection, and the visual rapid detection masks prepared in example 2 were worn separately, and significant color change could be observed on the detection line for how long it took to observe. The result shows that when the mask wearing time of the early patient reaches 55-60min, the color change of the detection line can be obviously observed; for middle and late stage patients, the color change of the detection line can be obviously observed when the mask is worn for 25-30 min.
According to the results, in order to achieve the purpose of effective detection, the wearing time of the mask is not less than 60min for the early-stage patient; for patients in middle and late stages, the mask is worn for no less than 30 min.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.
Claims (10)
1. A visual rapid detection mask for exhaled breath coronavirus is characterized by comprising a mask main body, a breather valve and a test strip;
the test strip is arranged on the breather valve and used for receiving the exhaled breath to be tested;
the test strip comprises a filter disc, a sample pad, a combination pad, a color development pad and a water absorption pad;
the sample pad, the combination pad, the color development pad and the water absorption pad are sequentially overlapped and are arranged on the filter sheet in a fitting manner;
the sample pad is positioned on the upper surface of the filter disc and is provided with a water absorption coating; the upper surface of the filter sheet faces the inner side of the mask main body;
the binding pad comprises colloidal gold coated with coronavirus N protein antibody;
the color development pad is positioned on the lower surface of the filter disc and is provided with a detection line and a quality control line, the detection line is close to one side of the combination pad, and the quality control line is close to one side of the water absorption pad; the detection line is coated with a coronavirus E protein antibody, and the quality control line is coated with an anti-coronavirus N protein antibody IgG;
the water absorption pad is positioned on the lower surface of the filter disc.
2. The visual rapid-detection mask of claim 1, wherein the mask body is selected from the group consisting of a disposable medical mask, a medical surgical mask, an industrial protective mask, and a civil protective mask.
3. The visual rapid-detection mask as claimed in claim 1, wherein the material of the water-absorbing coating of the sample pad comprises one or more of MOFs-HKUST-1, Cr-soc-MOF-1, NU-1500-Cr, activated carbon, porous silicon.
4. The visual rapid-detection mask according to claim 1, wherein the colloidal gold has a particle size of 10 to 50 nm.
5. The visual rapid-detection mask according to claim 1, wherein the coronavirus N protein antibody comprises rabbit anti-SARS Nsp3 antibody, rabbit MERS & SARS-CoV NP antibody;
the binding concentration of the coronavirus N protein antibody coated colloidal gold is 30-60 mu g/ml.
6. The visual rapid-detection mask according to claim 1, wherein said coronavirus E protein antibody comprises rabbit anti-SARS E protein antibody;
the coating concentration of the coronavirus E protein antibody is 1-5 mu g/cm.
7. The visual rapid-detection mask according to claim 1, wherein the anti-coronavirus E protein antibody IgG is goat anti-rabbit IgG monoclonal antibody;
the coating concentration of the anti-coronavirus E protein antibody IgG is 1-5 mu g/cm.
8. The method for preparing a visual rapid-detection mask according to any one of claims 1 to 7, comprising the steps of:
(1) coating the surface of the sample pad with the water-absorbing coating, wherein the thickness is 1-2 mm;
(2) coating the coronavirus N protein antibody on the colloidal gold, uniformly coating the colloidal gold coated with the coronavirus N protein antibody on the surface of the binding pad, and airing at room temperature;
(3) respectively engraving a detection line and a quality control line on the developing pad, coating a coronavirus E protein antibody on the detection line, coating an anti-coronavirus N protein antibody IgG on the quality control line, and drying at room temperature;
(4) sequentially adhering the sample pad prepared in the step (1), the bonding pad prepared in the step (2), the color development pad prepared in the step (3) and the water absorption pad on a filter sheet to prepare the test strip;
(5) taking the mask body provided with the breather valve, fixing the test strip prepared in the step (4) on the breather valve, and making the sample pad face the inner side of the mask, namely preparing the visual rapid detection mask for the exhaled coronary artery disease.
9. The method of claim 8, wherein step (1) comprises: mixing a copper nitrate solution and an ethanolamine solution, standing at room temperature for 24-36h, and performing suction filtration through an organic phase filter membrane to obtain a first film material; then reacting the first film material with an ethanol water solution of trimesic acid at normal temperature for 0.5-2h to obtain a second film material; a second thin film material is applied to the surface of the sample pad.
10. The method according to claim 8, wherein in the step (2), the method for preparing the colloidal gold comprises: taking chloroauric acid as a main raw material, and preparing the colloidal gold through a reduction reaction; the reducing agent is one or more selected from sodium citrate, tannic acid, ascorbic acid, white phosphorus, and sodium borohydride.
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| CN114223997A (en) * | 2021-12-28 | 2022-03-25 | 武汉大学 | Mask |
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