A highly safe novel coronavirus antigen detection kit

Abstract

The present invention discloses a highly secure novel coronavirus antigen detection kit comprising a test card, a swab, and a sample buffer. The sample buffer comprises the following components, measured by mass: 0.5-4 parts realgar, 0.5-4 parts borneol, 3-8 parts peppermint oil, 3-8 parts alum, 5-10 parts rosin, 5-10 parts tannic acid, 1-20 parts sucrose, 3-20 parts sodium chloride, 0.5-1.5 parts casein, 1-10 parts anionic surfactant, 0.5-3 parts sodium benzoate, 0.5-3 parts preservative, 0.01-0.03 parts chelating agent, 10-40 parts rice wine, and 700-1500 parts sterile water. This kit enhances safety while reducing the risk of virus transmission, protects low-level structural proteins, improves detection accuracy, and accelerates the chromatography speed of the sample buffer in the test card, shortening detection time.

Description

Technical Field

[0001] The present invention relates to the medical field, and in particular to a high-safety novel coronavirus antigen detection kit that reduces the risk of viral infection and effectively improves the safety of the detection process. Background Art

[0002] During the nucleic acid testing process, people usually need to put the throat swab or nasal swab into a buffer solution for preservation after use, and this process usually needs to be completed immediately. However, if the cotton swab contains the virus and enters the buffer solution, the splashing water can easily cause contamination to subsequent testing personnel and surrounding personnel and the environment. Currently, this buffer solution does not have the function of preventing this situation, and this route cannot be ruled out as one of the transmission routes.

[0003] Common technical means in the process of new coronavirus antigen testing include the latex method for new coronavirus antigen testing and the colloidal gold method for new coronavirus antigen testing. The latex method uses latex particles instead of colloidal gold particles. Although it reduces the production cost, there is no obvious difference in detection accuracy, and the color development speed of the latex method is still faster than that of the colloidal gold method.

[0004] Therefore, there is an urgent need for a high-safety novel coronavirus antigen detection kit that reduces the risk of viral transmission and effectively improves the safety of the detection process. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide a high-safety novel coronavirus antigen detection kit that reduces the risk of virus transmission and effectively improves the safety of the detection process.

[0006] The present invention provides a highly safe novel coronavirus antigen detection kit comprising a detection card, a swab, and a sample buffer;

[0007] The test card includes a sample pad, a conjugate pad, a reaction membrane, and absorbent paper;

[0008] The detection card is connected in sequence along the direction of sample chromatography, wherein the sample pad, conjugate pad, reaction membrane and absorbent paper are connected in sequence;

[0009] The reaction membrane is provided with a T line and a C line, wherein the T line contains a novel coronavirus nucleocapsid protein antibody or antigen-binding fragment, and the C line contains a ligand that specifically binds to a colloid-labeled quality control marker;

[0010] The sample buffer comprises the following components, calculated by mass fraction: 0.5-4 parts of realgar, 0.5-4 parts of borneol, 3-8 parts of peppermint oil, 3-8 parts of alum, 5-10 parts of rosin, 5-10 parts of tannic acid, 1-20 parts of sucrose, 3-20 parts of sodium chloride, 0.5-1.5 parts of casein, 1-10 parts of anionic surfactant, 0.5-3 parts of sodium benzoate, 0.5-3 parts of preservative, 0.01-0.03 parts of chelating agent, 10-40 parts of rice wine, and 700-1500 parts of sterile water.

[0011] The present invention provides a high-safety novel coronavirus antigen detection kit, wherein the sample buffer is composed of the following materials in parts by weight: 1-3 parts of realgar, 1-3 parts of borneol, 4-6 parts of peppermint oil, 4-6 parts of alum, 6-8 parts of rosin, 6-8 parts of tannic acid, 5-15 parts of sucrose, 5-15 parts of sodium chloride, 0.8-1.2 parts of casein, 3-7 parts of anionic surfactant, 1-2 parts of sodium benzoate, 1-2 parts of preservative, 0.015-0.025 part of chelating agent, 15-30 parts of rice wine, and 950-1200 parts of sterile water.

[0012] The present invention provides a high-safety novel coronavirus antigen detection kit, wherein the sample buffer is composed of the following materials in parts by weight: 2 parts of realgar, 2 parts of borneol, 5 parts of peppermint oil, 5 parts of alum, 7 parts of rosin, 7 parts of tannic acid, 10 parts of sucrose, 10 parts of sodium chloride, 1.1 parts of casein, 4 parts of anionic surfactant, 1.5 parts of sodium benzoate, 1.5 parts of preservative, 0.02 part of chelating agent, 20 parts of rice wine, and 1100 parts of sterile water.

[0013] The present invention provides a high-safety novel coronavirus antigen detection kit, wherein the sample buffer of the high-safety novel coronavirus antigen detection kit is configured as follows:

[0014] Step 1, preparing the first agent: crushing borneol and realgar, passing through a 200-mesh sieve, adding rice wine and continuing to grind, adding 20 parts of sterile water after each 1 minute of grinding and continuing to grind, and continuing to grind for 15 minutes until the fine powder completely floats in the liquid to form a suspension, and no more substances are dissolved or precipitated, thereby preparing the first agent;

[0015] Step 2, preparing a second agent: crushing alum, adding peppermint oil and the remaining sterile water, mixing, decocting the mixture over a low heat for 8 minutes, stirring 8 times clockwise and 8 times counterclockwise every 2 minutes, each stirring time not exceeding 16 seconds, passing through a 200-mesh sieve, and standing at 25 degrees for 2 hours to prepare the second agent;

[0016] Step 3, preparing the third agent: adding crushed rosin and tannic acid to the second agent, and concentrating the mixture under vacuum. The specific reaction conditions are: a concentration vacuum degree of -0.08 to -0.04 MPa, a concentration temperature of 68 to 84 degrees Celsius, and a relative density of the resulting concentrate at 60 degrees Celsius of 1.03 to 1.18. After standing for 4 hours, the first agent is added and the mixture is thoroughly stirred. The mixture is sieved through a 200-mesh sieve to finally prepare the third agent.

[0017] Step 4, preparing the fourth agent: adding sucrose, sodium chloride, casein, sodium benzoate, anionic surfactant, preservative, and chelating agent to the third agent and stirring thoroughly to prepare the fourth agent;

[0018] Step 5: Fill the fourth agent into a buffer sample tube and seal it.

[0019] The high-safety novel coronavirus antigen detection kit of the present invention is different from the prior art in that the present invention adds traditional Chinese medicine ingredients: realgar, alum, rosin, borneol, and peppermint oil to reduce the number of miscellaneous bacteria in the buffer solution and improve the accuracy of detection. Realgar inhibits skin fungi such as Staphylococcus aureus and human tuberculosis bacteria, thereby reducing the contamination of the buffer solution by the sample; alum can absorb certain pigments, protecting the characteristics of low-level structural proteins while not interfering with the cleavage of high-level structural proteins by anionic surfactants, thereby improving the success rate of antibody and antigen binding and making the detection results more accurate; rosin thickens the buffer solution while reducing the area of ​​splashing of the buffer solution after being impacted, thereby effectively preventing the overflow of the buffer solution containing the virus during monitoring, and at the same time, the rosin polyol esters are used to increase the speed of virus cleavage by anionic surfactants; borneol and peppermint oil can improve the pungent odor of realgar, alum and other organic solvents, avoiding coughing or sneezing in patients after being stimulated by the odor during the detection process, thereby reducing the possibility of virus transmission through droplets during the detection process.

[0020] The following is a further description of a high-safety novel coronavirus antigen detection kit of the present invention with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a schematic diagram of the experimental paper box of a high-safety novel coronavirus antigen detection kit;

[0022] Figure 2 This is a schematic diagram of the unfolding of the experimental paper box of a high-safety novel coronavirus antigen detection kit;

[0023] Figure 3 It is a statistical graph of the staining area of ​​each group;

[0024] Figure 4 It is a statistical chart of the waiting time of each group. DETAILED DESCRIPTION

[0025] like Figures 1 to 4 As shown, the present invention provides a highly safe novel coronavirus antigen detection kit, comprising a detection card, a swab, and a sample buffer;

[0026] The test card includes a sample pad, a conjugate pad, a reaction membrane, and absorbent paper;

[0027] The detection card is connected in sequence along the direction of sample chromatography, wherein the sample pad, conjugate pad, reaction membrane and absorbent paper are connected in sequence;

[0028] The reaction membrane is provided with a T line and a C line, wherein the T line contains a novel coronavirus nucleocapsid protein antibody or antigen-binding fragment, and the C line contains a ligand that specifically binds to a colloid-labeled quality control marker;

[0029] The sample buffer comprises the following materials in parts by weight: 0.5-4 parts of realgar, 0.5-4 parts of borneol, 3-8 parts of peppermint oil, 3-8 parts of alum, 5-10 parts of rosin, 5-10 parts of tannic acid, 1-20 parts of sucrose, 3-20 parts of sodium chloride, 0.5-1.5 parts of casein, 1-10 parts of anionic surfactant, 0.5-3 parts of sodium benzoate, 0.5-3 parts of preservative, 0.01-0.03 parts of chelating agent, 10-40 parts of rice wine and 700-1500 parts of sterile water.

[0030] The present invention provides a high-safety novel coronavirus antigen detection kit. Traditional Chinese medicine ingredients, including realgar, alum, rosin, borneol, and peppermint oil, are added to the buffer to reduce the number of miscellaneous bacteria in the buffer and improve the accuracy of detection. Realgar inhibits skin fungi such as Staphylococcus aureus and Mycobacterium tuberculosis, thereby reducing contamination of the buffer by the sample. Alum can absorb certain pigments, protecting the characteristics of low-level structural proteins while not interfering with the cleavage of high-level structural proteins by anionic surfactants, thereby improving the success rate of antibody and antigen binding and making the detection results more accurate. Rosin thickens the buffer and reduces the area of ​​splashing of the buffer after being impacted, thereby effectively preventing the overflow of the buffer containing the virus during monitoring. At the same time, the rosin polyol esters increase the speed of virus cleavage by the anionic surfactant. Borneol and peppermint oil can improve the pungent odor of realgar, alum, and other organic solvents, thereby preventing patients from coughing or sneezing due to odor stimulation during the detection process, thereby reducing the possibility of virus transmission through droplets during the detection process.

[0031] The interval between the T line (detection line) and the C line (quality control line) is 4 to 8 mm.

[0032] Wherein, the reaction membrane may be a nitrocellulose membrane.

[0033] Among them, the radius of the latex is 200nm.

[0034] The conjugate pad includes latex microspheres of goat anti-rabbit IgG, and the coating concentration is 0.5 mg / ml to 1.5 mg / ml.

[0035] Among them, the preparation method of the coupled goat anti-rabbit IgG latex microspheres is: using EDC and suflo-NHS to activate latex microspheres, mixing with goat anti-rabbit IgG, and covalently coupling. After sufficient reaction, blocking buffer is added for blocking. This technology is existing technology and will not be repeated here.

[0036] Pharmacological Description

[0037] Realgar: It has an inhibitory effect on common pathogenic skin fungi, such as common pyogenic cocci, intestinal pathogens, human and bovine tuberculosis bacteria, Clostridium smegmatis, and common fungi. Its water extract has varying degrees of inhibitory effect on Staphylococcus aureus, human tuberculosis bacteria, Proteus, Pseudomonas aeruginosa, and various skin fungi, and can effectively screen and control viruses and bacteria in buffer solutions.

[0038] Borneol: Its smell has strong penetrability and is beneficial for dispersing fire poison, removing wind and reducing phlegm. Combined with mint, it can effectively mask the strong irritating smell of realgar and alum. Especially for those with upper respiratory tract infections, it can effectively soothe throat pain, avoid stimulating patients to cough during nucleic acid testing, and reduce the risk of virus transmission.

[0039] Peppermint oil: It has a pungent and cool nature. Its volatile fragrance can dispel filth, clear the head, and has a strong sweating ability. It can treat wind-heat attacking the surface. The menthone it contains has a cool fragrance with a woody background that is not very long-lasting. As a neutral substance, it can effectively promote the dissolution of organic or fat-soluble substances. Together with borneol, it can improve the odor of the buffer solution, reduce coughing caused by odor stimulation during use, and thus reduce the probability of droplet transmission.

[0040] Alum: It has inhibitory effects on a variety of Gram-negative bacteria, Gram-positive bacteria, cocci and bacilli. At the same time, it can stabilize low-level structural proteins with tannic acid, preventing the anionic surfactant from continuing to destroy the low-level structural proteins after destroying the high-level structure of the viral protein. This can effectively improve the success rate of antibody capture and antigen binding, making the test data more accurate. At the same time, after alum is hydrolyzed, it has a certain adsorption effect on the pigment produced during the decoction process, thereby preventing the pigment from contaminating the test paper during the detection process.

[0041] Rosin: As one of the thickening components of the buffer solution, it can not only reduce and increase the viscosity of the buffer solution but also make the liquid surface of the buffer solution less likely to splash liquid droplets, thereby reducing the virus spillage caused by liquid droplets during the sample input process. At the same time, the rosin polyol esters contained in rosin can enhance the effect of anionic surfactants, thereby promoting faster virus lysis and allowing the buffer solution to reach a safe level more quickly.

[0042] Tannic acid: It has significant antibacterial properties, specifically by effectively inhibiting the formation of Staphylococcus aureus biofilms, thereby assisting realgar in controlling the number of bacteria in the buffer solution. At the same time, tannic acid can effectively inhibit the activity of structural proteins and promote the coagulation of structural proteins, which allows the viral antigens that are cleaved by anionic surfactants to maintain a certain state, avoiding the decrease in the number of antigens that can bind to antibodies due to the fragmentation of the viral structural proteins. This is not only beneficial to improving the accuracy of the test kit, but also can effectively control the number of viruses with transmission capabilities and improve the safety of the buffer solution.

[0043] Sodium benzoate: It is used to assist and enhance the antiseptic effect of Proclin 300, thereby facilitating the long-term preservation of the buffer solution. At the same time, sodium benzoate, as a cosolvent for low-molecular compounds, can effectively enhance the solubility of anionic surfactants in water, thereby improving the molecular movement in the buffer solution, thereby accelerating the chromatography speed of the buffer solution in the test paper, and accelerating the movement speed of the buffer solution in the test card under the action of the absorbent paper, thereby reducing the detection time and improving the detection efficiency.

[0044] As a preferred ratio of the sample buffer: the sample buffer of the high-safety novel coronavirus antigen detection kit of the present invention is composed of the following materials by weight:

[0045] 1-3 parts of realgar, 1-3 parts of borneol, 4-6 parts of peppermint oil, 4-6 parts of alum, 6-8 parts of rosin, 6-8 parts of tannic acid, 5-15 parts of sucrose, 5-15 parts of sodium chloride, 0.8-1.2 parts of casein, 3-7 parts of anionic surfactant, 1-2 parts of sodium benzoate, 1-2 parts of preservative, 0.015-0.025 parts of chelating agent, 15-30 parts of rice wine, and 950-1200 parts of sterile water.

[0046] The present invention can further enhance the formulation effect by optimizing the above-mentioned component ratio, improve the viscosity of the sample buffer, and optimize the proportions of anionic surfactant and sodium benzoate, so that the buffer improves detection safety while accelerating detection speed.

[0047] As a preferred ratio of the sample buffer: the sample buffer of the high-safety novel coronavirus antigen detection kit of the present invention is composed of the following materials by weight:

[0048] 2 parts of realgar, 2 parts of borneol, 5 parts of peppermint oil, 5 parts of alum, 7 parts of rosin, 7 parts of tannic acid, 10 parts of sucrose, 10 parts of sodium chloride, 1.1 parts of casein, 4 parts of anionic surfactant, 1.5 parts of sodium benzoate, 1.5 parts of preservative, 0.02 part of chelating agent, 20 parts of yellow rice wine, and 1100 parts of sterile water.

[0049] The present invention optimizes the ratio of the above components to achieve the most balanced ratio of alum and rosin, thereby improving the viscosity and safety of the sample buffer to a higher level and further improving the solubility of the sample buffer. In addition, the speed of the chromatography and diffusion process of the sample buffer on the test paper is accelerated, thereby reducing the waiting time required for detection.

[0050] The present invention provides a high-safety novel coronavirus antigen detection kit, wherein the sample buffer of the high-safety novel coronavirus antigen detection kit is configured as follows:

[0051] Step 1, preparing the first agent: crushing borneol and realgar, passing through a 200-mesh sieve, adding rice wine and continuing to grind, adding 20 parts of sterile water after each 1 minute of grinding and continuing to grind, and continuing to grind for 15 minutes until the fine powder completely floats in the liquid to form a suspension, and no more substances are dissolved or precipitated, thereby preparing the first agent;

[0052] The present invention uses the above-mentioned operation to add yellow rice wine and sterile water to the crushed borneol, realgar and licorice and continue to grind them, which can reduce the thermal changes and oxidation generated by the mineral medicine during grinding. For finer powders, the flying of powders can be effectively reduced as the grinding intensity increases. At the same time, since test paper needs to be used, reducing the size of the particles is conducive to improving the detection speed. Since the active ingredient of realgar is a mineral, only high fineness can be used to improve the medicinal properties. At the same time, yellow rice wine is added to help dissolve the medicinal components of fat-soluble substances, thereby maximizing the utilization of the medicinal value.

[0053] The grinding speed may be specifically 40 revolutions per minute, 20 revolutions clockwise and 20 revolutions counterclockwise, and the cycle repeats.

[0054] Step 2, preparing a second agent: crushing alum, adding peppermint oil and the remaining sterile water, mixing, decocting the mixture over a low heat for 8 minutes, stirring 8 times clockwise and 8 times counterclockwise every 2 minutes, each stirring time not exceeding 16 seconds, passing through a 200-mesh sieve, and standing at 25 degrees for 2 hours to prepare the second agent;

[0055] The present invention crushes mint and alum, the medicinal properties of which are difficult to extract, and blends them with sterile water before decocting, and helps dissolve the effective ingredients in the medicine during the stirring and decocting process to prepare the second medicament. In order to avoid caramelization of sugars in the traditional Chinese medicine caused by high temperature, which causes the color of the mixed liquid to darken, slow fire is used for heating to slow down the progress and degree of caramelization. At the same time, alum adsorbs pigments generated during the decocting process, and controls the generation of pigments in the traditional Chinese medicine during the decocting process and the adsorption and recovery after the generation, so that the color of the second medicament is more transparent, which is more convenient to display on a test paper later.

[0056] The temperature of the slow fire can be 80-120 degrees, preferably 90 degrees.

[0057] The invention sets the temperature of the slow fire to avoid caramelization of sugars and darkening the color of the solution.

[0058] Step 3, preparing the third agent: adding crushed rosin and tannic acid to the second agent, and concentrating the mixture under vacuum. The specific reaction conditions are: a concentration vacuum degree of -0.08 to -0.04 MPa, a concentration temperature of 68 to 84 degrees Celsius, and a relative density of the resulting concentrate at 60 degrees Celsius of 1.03 to 1.18. After standing for 4 hours, the first agent is added and the mixture is thoroughly stirred. The mixture is sieved through a 200-mesh sieve to finally prepare the third agent.

[0059] The present invention makes the solvent vapor discharge faster by means of reduced pressure concentration, while effectively reducing the decomposition of heat-sensitive substances while ensuring that the drug substance in the second agent is further dissolved. In particular, for drugs with large viscosity such as rosin, low-temperature evaporation can effectively prevent drug coking, thereby balancing the viscosity of the mixed solution and the content of the active ingredient of the drug in the solution to the greatest extent, further promoting the exudation of other medicinal materials while extracting the active ingredient of rosin, and effectively reducing the Maillard reaction of sugars and proteins in the mixed solution by lower temperature, and can also control the caramelization of sugars in the medicinal materials, thereby effectively reducing the content of brown macromolecular substances such as melanoidins or pseudo-melanin in the mixed solution, so that the color of the mixed solution is maintained at a lighter level, avoiding the unclear display of the test line of the test kit due to the dark color. Not only can the overall viscosity of the buffer solution be maintained at a high viscosity level, but also the solution ratio inside the buffer solution can be ensured to be uniform, avoiding insufficient local reaction after the sample is put into the buffer solution, thereby increasing the flow rate of the buffer solution and further reducing the detection time.

[0060] Step 4, preparing the fourth agent: adding sucrose, sodium chloride, casein, sodium benzoate, anionic surfactant, preservative, and chelating agent to the third agent and stirring thoroughly to prepare the fourth agent;

[0061] The present invention ensures the storage time of the buffer solution while improving the safety of the buffer solution after the sample is added by adding the necessary components required for the buffer solution. The necessary living environment for the organism is provided by adding sucrose, sodium chloride and casein, the virus is lysed by anionic surfactants to slow down its transmission ability, and the stability of the components in the buffer solution is improved by using chelating agents.

[0062] Among them, the chelating agent can be ethylenediaminetetraacetic acid. Since most nucleases and some proteases require Mg2+, it can also be used to remove the inhibitory effect of heavy metal ions on enzymes.

[0063] Wherein, the preservative may be Proclin 300.

[0064] Wherein, the anionic surfactant can be sodium lauryl sulfate.

[0065] Wherein, the sodium benzoate may be 0.1-0.3 g / L, preferably 0.2 g / L.

[0066] Step 5: Fill the fourth agent into a buffer sample tube and seal it.

[0067] Experimental data

[0068] Take the buffer solution and add red colorant, see Figure 1 Place the sampling tube of the mixed oil colorant in a white paper box with a side length of 10 cm. The middle of the paper box and the sampling tube can be detachably fixed. Put a metal ball with a mass of 4g into the buffer solution from 100mm above the sampling tube, then take out the sampling tube and unfold the paper box. Figure 2 The staining area was calculated after acquisition using an image instrument. Each group was repeated 50 times and the average staining area of ​​each group was calculated.

[0069] After adding the same sample, drop the buffer solution on the test card and start timing until the color of the C line and T line no longer changes, and record the waiting time. After 50 times for each group, statistics and calculate the average waiting time for each group.

[0070] The staining area and waiting time were recorded for the comparison group, control groups 1 and 2, and examples 1 to 3. It should be noted that the test card specifications and sizes of the control group and the example group are the same as those of the comparison group. The experimental data are as follows:

[0071] Comparison group:

[0072] This group used the Novel Coronavirus (2019-nCoV) Antigen Detection Kit (latex method), developed by Beijing Kingwolf Bioengineering Technology Co., Ltd. and manufactured by Aikon Biotechnology (Hangzhou) Co., Ltd., with national device registration number 20203400831. After unfolding the paper box, the image was collected and calculated using an imaging instrument, and the average staining area was 0.324 cm 2 At the same time, the recommended waiting time marked on this product is 15 to 20 minutes, and the average waiting time is 14.3 minutes.

[0073] The following is the control group. It should be noted that the preparation method of the buffer solution of the control group 1 is:

[0074] Step 1, preparing the first agent: crushing borneol and realgar, passing through a 200-mesh sieve, and adding yellow rice wine and 300 parts of sterile water to prepare the first agent;

[0075] Step 2, preparing a second agent: crushing alum, adding peppermint oil and the remaining sterile water, mixing, decocting the mixture over a low heat for 8 minutes, stirring 8 times clockwise and 8 times counterclockwise every 2 minutes, each stirring time not exceeding 16 seconds, passing through a 200-mesh sieve, and standing at 25 degrees for 2 hours to prepare the second agent;

[0076] Step 3, preparing the third agent: adding crushed rosin and tannic acid to the second agent, and concentrating the mixture under vacuum. The specific reaction conditions are: a concentration vacuum degree of -0.08 to -0.04 MPa, a concentration temperature of 68 to 84 degrees Celsius, and a relative density of the resulting concentrate at 60 degrees Celsius of 1.03 to 1.18. After standing for 4 hours, the first agent is added and the mixture is thoroughly stirred. The mixture is sieved through a 200-mesh sieve to finally prepare the third agent.

[0077] Step 4, preparing the fourth agent: adding sucrose, sodium chloride, casein, sodium benzoate, anionic surfactant, preservative, and chelating agent to the third agent and stirring thoroughly to prepare the fourth agent;

[0078] Step 5: Fill the fourth agent into a buffer sample tube and seal it.

[0079] The preparation method of the buffer solution of control group 2 is:

[0080] Step 1, preparing the first agent: crushing borneol and realgar, passing through a 200-mesh sieve, adding rice wine and continuing to grind, adding 20 parts of sterile water after each 1 minute of grinding and continuing to grind, and continuing to grind for 15 minutes until the fine powder completely floats in the liquid to form a suspension, and no more substances are dissolved or precipitated, thereby preparing the first agent;

[0081] Step 2, preparing a second agent: crushing alum, adding peppermint oil and the remaining sterile water, mixing, decocting the mixture over a low heat for 8 minutes, stirring 8 times clockwise and 8 times counterclockwise every 2 minutes, each stirring time not exceeding 16 seconds, passing through a 200-mesh sieve, and standing at 25 degrees for 2 hours to prepare the second agent;

[0082] Step 3, preparing the third agent: adding crushed tannic acid to the second agent, and concentrating the mixture under vacuum. The specific reaction conditions are: a concentration vacuum degree of -0.08 to -0.04 MPa, a concentration temperature of 68 to 84 degrees Celsius, and a relative density of the resulting concentrate at 60 degrees Celsius of 1.03 to 1.18. After standing for 4 hours, the first agent is added and stirred thoroughly. The mixture is sieved through a 200-mesh sieve to finally obtain the third agent.

[0083] Step 4, preparing the fourth agent: adding sucrose, sodium chloride, casein, sodium benzoate, anionic surfactant, preservative, and chelating agent to the third agent and stirring thoroughly to prepare the fourth agent;

[0084] Step 5: Fill the fourth agent into a buffer sample tube and seal it.

[0085] The following are examples. It should be noted that the preparation methods used for the buffer solutions of Examples 1-3 are:

[0086] Step 1, preparing the first agent: crushing borneol and realgar, passing through a 200-mesh sieve, adding rice wine and continuing to grind, adding 20 parts of sterile water after each 1 minute of grinding and continuing to grind, and continuing to grind for 15 minutes until the fine powder completely floats in the liquid to form a suspension, and no more substances are dissolved or precipitated, thereby preparing the first agent;

[0087] Step 2, preparing a second agent: crushing alum, adding peppermint oil and the remaining sterile water, mixing, decocting the mixture over a low heat for 8 minutes, stirring 8 times clockwise and 8 times counterclockwise every 2 minutes, each stirring time not exceeding 16 seconds, passing through a 200-mesh sieve, and standing at 25 degrees for 2 hours to prepare the second agent;

[0088] Step 3, preparing the third agent: adding crushed rosin and tannic acid to the second agent, and concentrating the mixture under vacuum. The specific reaction conditions are: a concentration vacuum degree of -0.08 to -0.04 MPa, a concentration temperature of 68 to 84 degrees Celsius, and a relative density of the resulting concentrate at 60 degrees Celsius of 1.03 to 1.18. After standing for 4 hours, the first agent is added and the mixture is thoroughly stirred. The mixture is sieved through a 200-mesh sieve to finally prepare the third agent.

[0089] Step 4, preparing the fourth agent: adding sucrose, sodium chloride, casein, sodium benzoate, anionic surfactant, preservative, and chelating agent to the third agent and stirring thoroughly to prepare the fourth agent;

[0090] Step 5: Fill the fourth agent into a buffer sample tube and seal it.

[0091] Control group 1: 1 part of realgar, 1 part of borneol, 4 parts of peppermint oil, 4 parts of alum, 6 parts of rosin, 6 parts of tannic acid, 5 parts of sucrose, 5 parts of sodium chloride, 0.8 parts of casein, 3 parts of anionic surfactant, 1 part of sodium benzoate, 1 part of preservative, 0.015 parts of chelating agent, 15 parts of rice wine, and 950 parts of sterile water.

[0092] After unfolding the carton, the image was collected and calculated by an imaging instrument, and the average staining area was 0.396 cm 2 The average waiting time for this group from the time the buffer solution was added until the color on the C line and T line no longer changed was 13.3 minutes.

[0093] Control group 2: 1 part of realgar, 1 part of borneol, 4 parts of peppermint oil, 4 parts of alum, 6 parts of tannic acid, 5 parts of sucrose, 5 parts of sodium chloride, 0.8 parts of casein, 3 parts of anionic surfactant, 1 part of sodium benzoate, 1 part of preservative, 0.015 parts of chelating agent, 15 parts of rice wine, and 950 parts of sterile water.

[0094] After unfolding the carton, the image was collected and calculated by an imaging instrument, and the average staining area was 0.712 cm 2 The average waiting time for this group from the time the buffer solution was added until the color on the C line and T line no longer changed was 10.9 minutes.

[0095] Example 1: 0.5 parts of realgar, 0.5 parts of borneol, 3 parts of peppermint oil, 3 parts of alum, 5 parts of rosin, 5 parts of tannic acid, 1 part of sucrose, 3 parts of sodium chloride, 0.5 parts of casein, 1 part of anionic surfactant, 0.5 parts of sodium benzoate, 0.5 parts of preservative, 0.01 parts of chelating agent, 10 parts of rice wine, and 700 parts of sterile water.

[0096] After unfolding the carton, the image was collected and calculated by an imaging instrument, and the average staining area was 0.304 cm 2 The average waiting time for this group from the time the buffer solution was added until the color on the C line and T line no longer changed was 9.3 minutes.

[0097] Example 2: 4 parts of realgar, 4 parts of borneol, 8 parts of peppermint oil, 8 parts of alum, 10 parts of rosin, 10 parts of tannic acid, 20 parts of sucrose, 20 parts of sodium chloride, 1.5 parts of casein, 10 parts of anionic surfactant, 1 part of sodium benzoate, 3 parts of preservative, 0.03 parts of chelating agent, 40 parts of rice wine, and 1500 parts of sterile water.

[0098] After unfolding the carton, the image was collected and calculated by an imaging instrument, and the average staining area was 0.242 cm 2 The average waiting time for this group from the time the buffer solution was added until the color on the C line and T line no longer changed was 10.4 minutes.

[0099] Example 3: 2 parts of realgar, 2 parts of borneol, 5 parts of peppermint oil, 5 parts of alum, 7 parts of rosin, 7 parts of tannic acid, 10 parts of sucrose, 10 parts of sodium chloride, 1.1 parts of casein, 4 parts of anionic surfactant, 1.5 parts of sodium benzoate, 1.5 parts of preservative, 0.02 parts of chelating agent, 20 parts of rice wine, and 1100 parts of sterile water.

[0100] After unfolding the carton, the image was collected and calculated by an imaging instrument, and the average staining area was 0.183 cm 2 The average waiting time for this group from the time the buffer solution was added until the color on the C line and T line no longer changed was 8.6 minutes.

[0101] According to the above experimental data, the following table is made:

[0102] See also Figure 3 、 4 The staining areas and waiting times of the comparison group, control group and embodiment group are shown in Table 1.

[0103]

[0104] Table 1

[0105] It can be seen from the above experimental data that the comparison group, the control group and the embodiment group are different in terms of staining area and waiting time. By comparing the comparison group, the control group and the embodiment group, the embodiment group increases the viscosity of the buffer solution to a certain extent by adding thickening substances such as rosin, and at the same time avoids the rosin from coking during the heating process by reducing pressure and concentrating, thereby causing the viscosity to decrease, thereby making the liquid splash less after being impacted, and thus the staining area is also smaller. As a buffer solution that may carry viruses, the fewer splashing droplets, the lower the possibility of virus transmission. In particular, for the control group 2 without adding rosin, its staining area is significantly improved relative to the control group 1 and the embodiment group, while the embodiment group has a significant reduction in staining area relative to the comparison group. In particular, Examples 2 and 3 can effectively reduce the probability of virus transmission through splashing droplets; and compared with the control group 1, in the absence of borneol and maleate, After the realgar is processed using a specific water-flying method, the particle diameter of the realgar increases, which causes an obstruction during the chromatography process on the test card, which makes the detection time of the control group 1 significantly longer than that of the other groups; and in Example 2, after the proportion of sodium benzoate is reduced while the proportion of other substances increases, the waiting time of Example 2 is significantly longer than that of Example 1 and Example 3. This is because in the absence of the solubilizing effect of ammonium benzoate, thickening substances such as rosin will reduce the solubility of the buffer solution, and relying solely on anionic surfactants cannot alleviate the reduced solubility. The reduction in solubility directly affects the chromatographic diffusion rate of the effective substances in the buffer solution on the test card, ultimately making the waiting time longer than the other two groups. At the same time, compared with the comparison group, the control group and the example group have added certain antibacterial components and protective components for low-level structural proteins to the necessary substances in the buffer solution, which also makes the detection results of the example group more accurate.

[0106] The embodiments described above are merely descriptions of preferred implementations of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should fall within the scope of protection determined by the claims of the present invention.

Claims

1. A highly safe novel coronavirus antigen detection kit, characterized by: Includes test card, swab, sample buffer; The sample buffer comprises the following components, calculated by mass fraction: 0.5-4 parts of realgar, 0.5-4 parts of borneol, 3-8 parts of peppermint oil, 3-8 parts of alum, 5-10 parts of rosin, 5-10 parts of tannic acid, 1-20 parts of sucrose, 3-20 parts of sodium chloride, 0.5-1.5 parts of casein, 1-10 parts of anionic surfactant, 0.5-3 parts of sodium benzoate, 0.5-3 parts of preservative, 0.01-0.03 parts of chelating agent, 10-40 parts of rice wine and 700-1500 parts of sterile water.

2. A high-safety novel coronavirus antigen detection kit according to claim 1, characterized in that The sample buffer solution is composed of the following materials in parts by weight: 1-3 parts of realgar, 1-3 parts of borneol, 4-6 parts of peppermint oil, 4-6 parts of alum, 6-8 parts of rosin, 6-8 parts of tannic acid, 5-15 parts of sucrose, 5-15 parts of sodium chloride, 0.8-1.2 parts of casein, 3-7 parts of anionic surfactant, 1-2 parts of sodium benzoate, 1-2 parts of preservative, 0.015-0.025 part of chelating agent, 15-30 parts of rice wine and 950-1200 parts of sterile water.

3. A high-safety novel coronavirus antigen detection kit according to claim 2, characterized in that The sample buffer is composed of the following materials in parts by weight: 2 parts of realgar, 2 parts of borneol, 5 parts of peppermint oil, 5 parts of alum, 7 parts of rosin, 7 parts of tannic acid, 10 parts of sucrose, 10 parts of sodium chloride, 1.1 parts of casein, 4 parts of anionic surfactant, 1.5 parts of sodium benzoate, 1.5 parts of preservative, 0.02 parts of chelating agent, 20 parts of rice wine and 1100 parts of sterile water.

4. A high-safety novel coronavirus antigen detection kit according to any one of claims 1 to 3, characterized in that: The sample buffer of the high-safety novel coronavirus antigen detection kit is prepared as follows: Step 1, preparing the first agent: crushing borneol and realgar, passing through a 200-mesh sieve, adding rice wine and continuing to grind, adding 20 parts of sterile water after each 1 minute of grinding and continuing to grind, and continuing to grind for 15 minutes until the fine powder completely floats in the liquid to form a suspension, and no more substances are dissolved or precipitated, thereby preparing the first agent; Step 2, preparing a second agent: crushing alum, adding peppermint oil and the remaining sterile water, mixing, decocting the mixture over a low heat for 8 minutes, stirring 8 times clockwise and 8 times counterclockwise every 2 minutes, each stirring time not exceeding 16 seconds, passing through a 200-mesh sieve, and standing at 25 degrees for 2 hours to prepare the second agent; Step 3, preparing the third agent: adding crushed rosin and tannic acid to the second agent, and concentrating the mixture under vacuum. The specific reaction conditions are: a concentration vacuum degree of -0.08 to -0.04 MPa, a concentration temperature of 68 to 84 degrees Celsius, and a relative density of the resulting concentrate at 60 degrees Celsius of 1.03 to 1.

18. After standing for 4 hours, the first agent is added and the mixture is thoroughly stirred. The mixture is sieved through a 200-mesh sieve to finally prepare the third agent. Step 4, preparing the fourth agent: adding sucrose, sodium chloride, casein, sodium benzoate, anionic surfactant, preservative and chelating agent to the third agent and stirring thoroughly to prepare the fourth agent; Step 5: Fill the fourth agent into a buffer sample tube and seal it.

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