Multiplex Viral Antigen Detection Kit and Its Application

Abstract

The present application discloses a kit for combined detection of multiple virus antigens and an application method thereof. The application method includes: placing the collected sample in an extraction tube; squeezing the bottle body of the extraction tube multiple times and standing for a preset time period to obtain a sample solution; wherein, the extraction tube contains a sample extraction solution that can simultaneously process multiple antigens; dropping the sample solution on multiple preset test strips and waiting for the antigens in the sample solution to react with the antibodies on the test strips; after the antigens in the sample solution react with the antibodies on the test strips, obtaining a test result based on the quality control line and the detection line set on the test strips. In the present application, dropping the same sample solution on different test strips can quickly determine the antigens present in the sample, and different test strips do not affect each other in terms of reaction and color development, improving the accuracy. Therefore, the detection efficiency of the virus is improved.

Description

Technical Field

[0001] The present application relates to the field of medical testing technologies, and particularly to a kit for joint detection of multiple virus antigens and its application. Background Art

[0002] As more and more viruses cause diseases with similar symptoms, it is impossible to diagnose the virus causing the disease only through symptoms, and antigen detection can only be performed based on the doctor's guess for further confirmation.

[0003] By detecting virus antigens, the virus causing the disease can be identified, enabling doctors to provide targeted treatment and improve the treatment effect. However, currently, when performing antigen detection on viruses causing diseases with similar symptoms, it is necessary to determine several viruses based on symptoms, then prepare different samples according to different viruses, and perform detection through the samples and the corresponding viruses, resulting in low detection speed. Or, different antigens are detected using the same test strip and the same sample. When detecting multiple antigens on the same test strip, since the antigens of viruses causing diseases with similar symptoms are also similar, there may be a situation where antibodies misidentify antigens, leading to inaccurate detection. That is, currently, when performing antigen detection on multiple viruses, the detection efficiency of viruses is low. Summary of the Invention

[0004] The main purpose of the present application is to provide a kit for joint detection of multiple virus antigens and its application, aiming to solve the technical problem of low detection efficiency of viruses when performing antigen detection on multiple viruses in the prior art.

[0005] To achieve the above object, the present application provides a kit for joint detection of multiple virus antigens, including:

[0006] A base on which a plurality of test strips are provided;

[0007] An extraction tube including a bottle body, with a detachable upper cover provided at the opening of the bottle body. An inner plug is provided between the upper cover and the bottle body. A dropper opening is provided at the top of the upper cover for dripping the sample, and a detachable top cover is provided at the dropper opening. The extraction tube is used to process the sample and drip the sample onto the test strip;

[0008] A test strip, each test strip including a sample pad, a conjugate pad, a chromatographic membrane, and an absorbent pad provided on a PVC bottom plate. A test line and a control line are provided on the chromatographic membrane. Antibodies for detecting viruses are coated on the test line, and goat anti-mouse IgG antibodies are coated on the control line. The multiple test strips are fixedly provided on the base 10. Optionally, the sample extraction solution includes a buffer and a lysis surfactant, wherein the formulation ratio of the buffer to the surfactant is 0.01 mol / L PBS and a 0.1% SDS solution, with a pH of 7.4.

[0009] Optionally, a conjugate pad is provided on one side of the chromatographic membrane, and different colloidal gold-labeled virus antibodies are distributed on the conjugate pads of different test strips, wherein the virus antibodies are novel coronavirus antibodies, influenza A / B virus antibodies, and respiratory syncytial virus antibodies.

[0010] Optionally, an absorbent pad is provided on the other side of the chromatographic membrane, and a sample pad is provided on the side of the conjugate pad away from the absorbent pad.

[0011] Optionally, the sample pad is a first glass cellulose membrane dried after being soaked in a preset sample pad treatment solution, and the sample pad treatment solution includes a buffer solution, a hydrophilic surfactant, and casein. The formulation ratio of the buffer solution, the surfactant, and the casein is PBS at 0.01 mol / L, Tween-20 solution at a concentration of 0.4%, and casein at a concentration of 0.5%, and the pH is 7.4.

[0012] Optionally, the conjugate pad is a second glass cellulose membrane dried after being soaked in a preset conjugate pad pretreatment solution, and the conjugate pad pretreatment solution includes a buffer agent, a surfactant, casein, and a preservative. The formulation of the conjugate pad pretreatment solution is PB at 0.01 mol / L, casein at a concentration of 0.3%, Tween-20 at a concentration of 0.5%, and PC300 preservative at a concentration of 0.01%, and the pH value is 7.4.

[0013] Optionally, the virus antibodies coated on the different detection lines on the chromatographic membranes are different, and a quality control line of the same goat anti-mouse IgG antibody is also coated on each chromatographic membrane.

[0014] Optionally, the chromatographic membrane includes an NC membrane. The detection line is drawn on the NC membrane with a coating buffer solution containing a virus antibody, and the quality control line is drawn on the NC membrane with a coating buffer solution containing a quality control antibody. The coating buffer solution includes a buffer agent, trehalose, and a preservative. The formulation ratio of the buffer agent, the trehalose, and the preservative is PB buffer at 0.01 mol / L, trehalose at a concentration of 1%, and PC300 preservative at a concentration of 0.01%.

[0015] This application also provides an application method for a multi-virus antigen combined detection kit. The application method for the multi-virus antigen combined detection kit includes:

[0016] Place the collected sample in an extraction tube;

[0017] Squeeze the bottle body of the extraction tube multiple times and let it stand for a preset period of time to obtain a sample solution;

[0018] Drop the sample liquid onto a variety of preset test strips and wait for the antigen in the sample liquid to react with the antibody on the test strip;

[0019] After the antigen in the sample liquid reacts with the antibody on the test strip, obtain a test result based on the quality control line and the detection line set on the test strip.

[0020] This application provides a kit for the combined detection of multiple virus antigens and its application. Compared with the prior art in which the detection efficiency of viruses is low when detecting antigens of multiple viruses, in this application, the collected sample is placed in an extraction tube; the bottle body of the extraction tube is squeezed multiple times and left stationary for a preset period of time to obtain a sample liquid; wherein, the extraction tube contains a sample extraction liquid for simultaneously processing multiple antigens; drop the sample liquid onto a variety of preset test strips and wait for the antigen in the sample liquid to react with the antibody on the test strip; after the antigen in the sample liquid reacts with the antibody on the test strip, obtain a test result based on the quality control line and the detection line set on the test strip. In this application, by placing the sample in an extraction tube, using the extraction liquid in the extraction tube that can simultaneously process multiple antigens to expose the nucleoprotein of the antigen in the sample to obtain a sample liquid, and then dropping the sample liquid onto different test strips respectively, and through the reaction of the antigen in the sample liquid with the antibody on the test strip to obtain a test result, that is, in this application, dropping the same sample liquid onto different test strips can quickly determine the antigens present in the sample, and different test strips will not affect the reaction and color development between each other, improving the accuracy, and thus improving the detection efficiency of viruses. Description of the Drawings

[0021] The drawings here are incorporated into the specification and form a part of this specification, showing embodiments consistent with this application and used together with the specification to explain the principles of this application.

[0022] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, for those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0023] Figure 1 It is a schematic flow chart of the application method of the kit for the combined detection of multiple virus antigens of this application;

[0024] Figure 2 It is a schematic structural diagram of the test strip of this application;

[0025] Figure 3 It is a schematic structural diagram of the base of the test strip of this application;

[0026] Figure 4 This is a schematic structural diagram of the test strip cover of the present application;

[0027] Figure 5 This is a schematic structural diagram of the extraction tube of the present application;

[0028] Figure 6 This is an application schematic diagram of the application method of the multi-virus antigen combined detection kit of the present application.

[0029] Explanation of the reference numerals in the attached drawings:

[0030] Label Name Label Name 1 Sample pad 2 Conjugate pad 3 Chromatographic membrane 4 Test line 5 Control line 6 Absorbent pad 100 Extraction tube 10 Base 11 Test strip cover 12 Observation window 13 Sample application hole 14 Top cover 15 Upper cover 16 Inner plug 17 Bottle body

[0031] The realization of the purpose, functional features and advantages of the present application will be further described in conjunction with the embodiments with reference to the accompanying drawings. Detailed implementation manners

[0032] It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0033] In the present application, a multi-virus antigen combined detection kit and its application are proposed, that is, the kit. Refer to Figures 1 to 6 , Figure 1 This is a schematic flow diagram of the application method of the multi-virus antigen combined detection kit of the present application; Figure 2 This is a schematic structural diagram of the test strip of the present application; Figure 3 This is a schematic structural diagram of the base of the present application; Figure 4 This is a schematic structural diagram of the test strip cover of the present application; Figure 5 This is a schematic structural diagram of the extraction tube of the present application; Figure 6 This is an application schematic diagram of the application method of the multi-virus antigen combined detection kit of the present application.

[0034] In the embodiments of the present application, the kit includes:

[0035] A base 10, on which a plurality of test strips are arranged;

[0036] An extraction tube 100, the extraction tube 100 includes a bottle body 17, a detachable upper cover 15 is arranged at the opening of the bottle body 17, an inner plug 16 is arranged between the upper cover 15 and the bottle body 17, a liquid dropping port is arranged at the top of the upper cover for dropping samples, a detachable top cover 14 is arranged at the liquid dropping port, and the extraction tube 100 is used for sample treatment and dropping the sample onto the test strip;

[0037] Test strip, each test strip includes a sample pad 1, a conjugate pad 2, a chromatographic membrane 3, and an absorbent pad 6 disposed on a PVC bottom plate. The chromatographic membrane 3 is provided with a test line 4 and a control line 5. The test line 4 is coated with an antibody for detecting a virus, and the control line 5 is coated with a goat anti-mouse IgG antibody. The multiple test strips are fixedly arranged on the base 10.

[0038] In an embodiment of the present application, since the antigens of viruses causing diseases with similar symptoms are similar. For example, the antigens of novel coronavirus, influenza A / B virus, and respiratory syncytial virus. Setting the antibodies of multiple viruses on the same test strip may result in detection errors. For example, a test strip is provided with antibodies of virus A and virus B, and the test line 4 of virus A is located before the test line 4 of virus B. If the sample contains virus B, virus B will pass through the test line 4 of virus A and then reach the test line 4 of virus B. When virus B passes through the test line 4 of virus A, some antigens of virus B may react with the antibody of virus A, causing the test line 4 of virus A to change color, and the test line 4 of virus B will also produce color. At this time, it will be misjudged that the sample contains a small amount of virus A and a large amount of virus B, thus causing misdetection. In this embodiment, a test line 4 is provided on the chromatographic membrane 3, and the test line 4 is coated with an antibody of a virus. It should be noted that the antibodies coated on the test line 4 of each chromatographic membrane 3 come from different pathogens. That is, by setting it like this, it can be avoided that the antibodies on the test strip recognize the wrong antigen, thereby avoiding misdetection.

[0039] In an embodiment of the present application, a plug 16 is provided between the body 17 of the sample extraction tube and the upper cover 15. By setting it like this, the sealing performance of the body 17 can be improved, and it can be avoided that the sample extraction liquid set in the body 17 overflows during transportation, resulting in leakage and contamination of the sample extraction liquid.

[0040] The top end of the upper cover 15 is provided with a dropper opening for dropping the sample, and a removable top cover 14 is provided at the dropper opening. The extraction tube is used to process the sample and drop the sample onto the test strip. When dropping the sample, the extraction tube can be inverted, the top cover 14 can be removed, and the sample can be directly dropped.

[0041] The plug 16 is set in a funnel shape, the thin end of the plug 16 is located on the side of the upper cover 15, and a sample extraction liquid is provided in the body 17.

[0042] In an embodiment of the present application, when dropping the sample extraction liquid onto the test strip, the plug needs to be taken out.

[0043] In an embodiment of the present application, the inner plug 16 is set in a funnel shape, that is, the diameter of one end of the inner plug 16 is large, and the diameter of the other end is small. The end with the large diameter of the inner plug 16 is close to the bottle body 17, and the end with the small diameter of the inner plug 16 is close to the upper cover 15. With this setting, when the extraction tube 100 is poured, the sample extraction liquid in the bottle body 17 can be intercepted by the inner plug 16, avoiding the leakage of the sample extraction liquid.

[0044] Among them, the thin mouth end of the inner plug 16 can be set in a pressure valve shape.

[0045] In an embodiment of the present application, if the thin mouth end of the inner plug 16 is set in a pressure valve shape, that is, pressure is applied to the end with the large diameter of the inner plug 16, and the end with the small diameter of the inner plug 16 will be opened. With this setting, when dropping the sample liquid onto the test strip through the extraction tube 100, that is, when the bottle body 17 is inverted, the sample liquid will not drip freely and contaminate the test kit.

[0046] The sample extraction liquid includes a buffer solution and a lysis surfactant. Among them, the formulation ratio of the buffer solution to the surfactant is PBS at 0.01 mol / L and an SDS solution with a concentration of 0.1%, and the pH is 7.4.

[0047] Among them, mole is the unit of amount of substance.

[0048] Among them, the lysis surfactant can be used to lyse the virus, inactivate the virus, and expose the nucleoprotein of the virus.

[0049] Among them, the buffer solution can be used to adjust the acid-base environment of the sample extraction liquid and maintain the acid-base environment.

[0050] Among them, SDS (Sodium dodecyl sulfate) can separate proteins from DNA.

[0051] In an embodiment of the present application, a lysis surfactant and a buffer solution are added to the sample extraction liquid. With this setting, the acid-base environment in the sample extraction liquid can be maintained stable by the buffer solution, enabling the lysis surfactant to exert its maximum effect. The virus is lysed by the lysis surfactant to inactivate the virus, which can avoid the spread of pathogens and cause harm to users. It can also completely expose the nucleoprotein of the virus in the sample extraction liquid, enabling the antibody on the detection line 5 to better recognize the antigen.

[0052] In an embodiment of the present application, the sample extraction liquid is composed of a PBS buffer solution at 0.01 mol / L and an SDS lysis surfactant with a concentration of 0.1%, and the pH is 7.4. With this configuration, it can be avoided that SDS does not completely lyse the virus, and it can also be avoided that the SDS concentration is too high and lyses the nucleoprotein of the virus.

[0053] In one embodiment of the present application, the PBS buffer can maintain the acid-base environment in the sample extract, can also adjust the osmotic pressure of the sample extract, and adjust the ion concentration at a certain intensity, so that the sample extract can extract more available samples from the collected comprehensive sample. The concentration of the PBS buffer is set to 0.01 mol / L to ensure that a sufficient amount of samples can be extracted.

[0054] A binding pad 2 is provided on one side of the chromatography membrane 3, and different colloidal gold-labeled virus antibodies are distributed on the binding pads 2 of different test strips. Among them, the virus antibodies are novel coronavirus antibodies, influenza A / B virus antibodies, and respiratory syncytial virus antibodies.

[0055] It should be noted that the antibodies used are all directed against the nucleoproteins (NP) of the novel coronavirus / influenza A / B virus / respiratory syncytial virus respectively. The nucleoproteins have high conservation and good specificity, which not only ensures the compatibility of the product but also ensures the specificity of the product and avoids cross-reactions.

[0056] It should be noted that two test lines are provided on the test strip labeled with influenza A / B virus antibodies to distinguish between influenza A virus and influenza B virus.

[0057] In one embodiment of the present application, different colloidal gold-labeled virus antibodies are provided on the binding pads 2 of different test strips. With this setting, the nucleoprotein (antigen) of the virus can first bind to the colloidal gold-labeled virus antibody on the binding pad 2, and then pass through the chromatography membrane to bind to the virus antibody on the test line, forming a sandwich structure, thereby detecting the virus.

[0058] In one embodiment of the present application, the antibodies used in the kit are all directed against the nucleoproteins (NP) of the novel coronavirus / influenza A / B virus / respiratory syncytial virus respectively. The nucleoproteins have high conservation and good specificity, which not only ensures the compatibility of the product but also ensures the specificity of the product and avoids cross-reactions.

[0059] In one embodiment of the present application, the method for labeling virus antibodies with colloidal gold can be as follows: adjust the colloidal gold solution to an appropriate pH value. In different colloidal gold solutions, add novel coronavirus antibodies, influenza A / B virus antibodies, and respiratory syncytial virus antibodies respectively. After stirring and mixing for a period of time, add a blocking agent, stir and mix for a period of time, centrifuge and concentrate, remove the supernatant, and dilute the precipitate with a colloidal gold diluent to the required concentration. Specifically, the formula of the colloidal gold diluent is: 0.01M PB + 5% sucrose + 5% Tre + 1% PEG6000 pH 7.4. The colloidal gold diluent is mainly composed of a buffer solution, sugars, and organic polymers. Among them, the buffer solution provides a stable reaction pH environment, and the sugars and organic polymers play a role in protecting the antibody activity and extending the shelf life.

[0060] On the other side of the chromatographic membrane 3, a water-absorbing pad 6 is provided, and a sample pad 1 is provided on the side of the binding pad 2 away from the water-absorbing pad 6.

[0061] In an embodiment of the present application, a sample pad 1 is provided on the binding pad 2. The sample pad 1 is used to receive the sample liquid dropped from the extraction tube 100. That is, after the sample liquid is prepared in the extraction tube 100, the sample liquid is dropped on the sample pad 1 and chromatographed from the sample pad 1 to the binding pad 2. If the sample liquid contains an antigen, it can bind to the specific antibody labeled with colloidal gold on the binding pad to form an antigen-antibody colloidal gold complex.

[0062] In an embodiment of the present application, a blotting paper is provided on the chromatographic membrane 3, and the blotting paper is provided on the opposite side of the sample pad 1. Such a setting can suck the sample liquid dropped on the sample pad 1 by the blotting paper and continue to move toward the test line 4. If the sample liquid contains an antigen, the antigen-antibody colloidal gold complex can be captured by the corresponding antibody on the test line to form a test line band. The sample pad 1 is set as a first glass cellulose membrane soaked in a preset sample pad 1 treatment liquid and then dried. The sample pad 1 treatment liquid includes a buffer solution, a hydrophilic surfactant, and casein. Among them, the formulation ratio of the buffer solution, the surfactant, and the casein is PBS at 0.01 mol / L, a Tween-20 solution with a concentration of 0.4%, and casein with a concentration of 0.5%.

[0063] Among them, the hydrophilic surfactant can improve the hydrophilicity of the material and facilitate chromatography.

[0064] Among them, casein can reduce the residual target protein and improve sensitivity.

[0065] In an embodiment of the present application, the first glass cellulose membrane is used as the material of the sample pad 1. Such a setting can utilize the characteristics of the first glass cellulose membrane with small resistance and high filtration efficiency to increase the chromatography effect of the sample liquid, reduce the resistance to the nucleoprotein of the virus, and enable the nucleoprotein of the virus to pass through chromatography to the greatest extent.

[0066] In an embodiment of the present application, by immersing the first cellulose acetate membrane in a sample pad 1 treatment solution with a formulation ratio of buffer solution, surfactant, and casein of PBS at 0.01 mol / L, Tween-20 solution at a concentration of 0.4%, and casein at a concentration of 0.5%, the sample pad 1 treatment solution can be completely immersed in the first cellulose acetate membrane. Through casein at a concentration of 0.5%, the nucleoprotein of the virus can be completely chromatographed onto the conjugate pad 2, and the hydrophilicity of the first cellulose acetate membrane can be changed through the hydrophilic surfactant, enabling the sample pad to better absorb the sample solution carrying the nucleoprotein. During the chromatography process of the sample pad 1, the buffer solution of PBS at 0.01 mol / L is used to maintain the acid-base environment of the sample solution and avoid sudden changes in the acid-base environment in the sample solution, which may damage the nucleoprotein.

[0067] Specifically, the production of the sample pad 1 can be achieved by cutting the cellulose acetate membrane into strips with a specification of 20.0 mm × 300.0 mm, soaking and treating it with the sample pad 1 treatment solution. After completion, it is transferred to an oven and dried at 45°C ± 2°C for 24 ± 2 hours.

[0068] The conjugate pad 2 is set to be a second cellulose acetate membrane that is soaked in a preset conjugate pad 2 pretreatment solution and then dried. The conjugate pad 2 pretreatment solution includes a buffer, a surfactant, casein, and a preservative. Among them, the formulation of the conjugate pad 2 pretreatment solution is PB at 0.01 mol / L, casein at a concentration of 0.3%, Tween-20 at a concentration of 0.5%, PC300 preservative at a concentration of 0.01%, and the pH value is 7.4.

[0069] Among them, PB is a phosphate buffer solution.

[0070] It should be noted that the preparation method of the conjugate pad 2 is: spraying the colloidal gold label on the second cellulose acetate membrane treated with the conjugate pad 2 pretreatment solution, and waiting for the second cellulose acetate membrane to dry to obtain the colloidal gold conjugate pad 2.

[0071] In an embodiment of the present application, by immersing the second cellulose acetate membrane in a conjugate pad 2 pretreatment solution with a formulation ratio of buffer, surfactant, casein, and preservative of PB at 0.01 mol / L, casein at a concentration of 0.3%, Tween-20 at a concentration of 0.5%, PC300 preservative at a concentration of 0.01%, and the pH value of 7.4, the conjugate pad 2 can be prevented from deteriorating during storage by the PC300 preservative at a concentration of 0.01%, which may affect the accuracy of the detection.

[0072] In an embodiment of the present application, the pH value of the sample solution needs to be set to 7.4, that is, the same as the acidity and alkalinity in the sample extraction solution, in order to maintain the integrity and independence of the nucleoprotein of the virus in the sample solution.

[0073] The virus antibodies coated on the detection lines 4 on different chromatographic membranes 3 are different, and a quality control line 5 with the same goat anti-mouse IgG antibody is also coated on each chromatographic membrane 3.

[0074] In an embodiment of the present application, the goat anti-mouse IgG antibody in the quality control line 5 is used to verify the validity of the detection result. If a red band appears on the quality control line 5, it indicates that the detection result is valid. If the quality control line 5 does not show color, it indicates that the detection result is invalid.

[0075] The chromatographic membrane 3 includes an NC membrane. The detection line 4 is drawn on the NC membrane with a coating buffer containing a virus antibody, and the quality control line 5 is drawn on the NC membrane with a coating buffer containing a quality control antibody. The coating buffer includes a buffer, trehalose, and a preservative. Among them, the formulation ratio of the buffer, trehalose, and preservative is 0.01 mol / L PB buffer, 1% trehalose, and 0.01% PC300 preservative.

[0076] In an embodiment of the present application, the chromatographic membrane 3 is obtained by coating, drawing lines, and drying an NC membrane. Specifically, the NC membrane is placed at a fixed position on the membrane drawing instrument platform, and the coating buffer is started to draw lines. The coated NC membranes should be numbered one by one, and the two ends, uneven width, breakpoints, and damaged parts of the test line and the quality control line should be marked with a pen in time. After membrane drawing, the NC membranes are arranged in order, facing up, and placed in an oven at a temperature of 45°C ± 2°C for 72 ± 2 hours. In an embodiment of the present application, the preparation method of the coating buffer can be 0.01M PB + 1% trehalose + 0.01% PC300, which mainly consists of a buffer, trehalose, and a preservative. The buffer provides a stable pH environment, trehalose plays a role in protecting the antibody activity and extending the validity period, and the preservative is used to prevent deterioration. Prepare 200 mL of buffer according to the above formula and adjust the pH to 7.4. Add a detection antibody to the prepared coating buffer to obtain a coating buffer with a detection antibody, where the detection antibody is the antibody of the virus to be detected. Add a quality control antibody to the prepared coating buffer to obtain a coating buffer with a quality control antibody, where the quality control antibody is a goat anti-mouse IgG antibody.

[0077] In an embodiment of the present application, a coating buffer with a quality control antibody is drawn on the chromatographic membrane 3 to detect the validity of the detection, and a coating buffer with a detection antibody is drawn on the chromatographic membrane 3 to detect the virus antigen. If the quality control line 5 shows color and the detection line 4 also shows color, it indicates that the sample solution contains the virus corresponding to the detection.

[0078] Further, based on the above embodiments in the present application, another embodiment of the present application is provided. In this embodiment, a test strip cover 11 is further provided on the base 10. A sample adding hole 13 and an observation window 12 are provided on the test strip cover 11. The sample adding hole 13 is located on the side of the test strip cover 11 away from the absorbent pad 6, and the observation window 12 is provided at the chromatography membrane for observing the quality control line 5 and the test line 4.

[0079] Further, based on the above embodiments in the present application, another embodiment of the present application is provided. In this embodiment, a method for applying a multi-virus antigen combined detection kit includes:

[0080] Step S10, placing the collected sample in an extraction tube;

[0081] Step S20, squeezing the bottle body of the extraction tube multiple times and standing for a preset time period to obtain a sample solution;

[0082] Wherein, the extraction tube includes a sample extraction solution capable of simultaneously processing multiple antigens;

[0083] Step S30, dropping the sample solution onto multiple preset test strips and waiting for the antigens in the sample solution to react with the antibodies on the test strips;

[0084] Step S40, after the antigens in the sample solution react with the antibodies on the test strips, obtaining a test result based on the quality control line and the test line set on the test strips.

[0085] Wherein, the type of the collected sample can be a nasal swab, a nasopharyngeal swab or an oropharyngeal swab.

[0086] In this embodiment, the collected sample is placed in an extraction tube, and the sample extraction solution in the extraction tube lyses the virus in the sample, thereby exposing the nucleoprotein in the sample. It should be noted that in order to wash the sample collected on the swab more thoroughly, the bottle body of the extraction tube can be squeezed multiple times.

[0087] In this embodiment, the sample extraction solution can simultaneously lyse multiple viruses, exposing the nucleoproteins of multiple viruses.

[0088] In this embodiment, the sample solution is dropped through the sample adding hole onto different test strips, so that the antibodies in different test strips recognize the corresponding nucleoproteins, that is, the antibodies in different test strips recognize the corresponding antigens, and the sample solution is continuously attracted to the quality control line and the test line through the absorbent pad in the test strips.

[0089] It should be noted that after dropping the sample solution into the sample adding hole, a reaction time needs to be left for the sample solution and the test strip to improve the credibility of the test result.

[0090] The present application provides a kit for jointly detecting multiple virus antigens and its application. Compared with the prior art where the detection efficiency of viruses is low when detecting antigens of multiple viruses, in the present application, the collected sample is placed in an extraction tube; the body of the extraction tube is squeezed multiple times and left static for a preset period of time to obtain a sample solution; wherein, the extraction tube contains a sample extraction solution that can simultaneously process multiple antigens; the sample solution is dropped on multiple preset test strips, and the antigens in the sample solution are waited to react with the antibodies on the test strips; after the antigens in the sample solution react with the antibodies on the test strips, based on the quality control line and the detection line set on the test strips, a detection result is obtained. In the present application, by placing the sample in the extraction tube, the nucleoproteins of the antigens in the sample are exposed through the extraction solution in the extraction tube that can simultaneously process multiple antigens to obtain a sample solution, and then the sample solution is respectively dropped on different test strips. Through the reaction of the antigens in the sample solution with the antibodies on the test strips, a detection result is obtained. That is, in the present application, dropping the same sample solution on different test strips can quickly determine the antigens present in the sample, and different test strips will not affect the reaction and color development between each other, improving the accuracy. Therefore, the detection efficiency of viruses is improved.

[0091] It should be noted that in this text, the terms "include", "comprise" or any other variant thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements not only includes those elements, but also includes other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. Without further limitation, an element defined by the statement "including one..." does not exclude the existence of another identical element in the process, method, article or device including that element.

[0092] The serial numbers of the above-mentioned embodiments of the present invention are only for description and do not represent the superiority or inferiority of the embodiments.

[0093] Through the description of the above embodiments, those skilled in the art can clearly understand that the above-mentioned embodiment methods can be implemented by means of software plus a necessary general hardware platform. Of course, they can also be implemented by hardware, but in many cases the former is a better implementation method. Based on such an understanding, the technical solution of the present invention, in essence, or the part that makes a contribution to the prior art, can be embodied in the form of a software product. This computer software product is stored in a storage medium (such as ROM / RAM, magnetic disk, optical disk) and includes several instructions for causing a terminal device (which can be a mobile phone, computer, server, air conditioner, or network device, etc.) to execute the methods described in various embodiments of the present invention.

[0094] The above are only the preferred embodiments of the present invention, and do not limit the patent scope of the present invention accordingly. Any equivalent mechanism or equivalent process transformation made by using the content of the specification and drawings of the present invention, or directly or indirectly applied in other related technical fields, shall be similarly included in the patent protection scope of the present invention.

Claims

1. A kit for the combined detection of multiple virus antigens, characterized in that, Comprising: A base on which a plurality of test strips are provided; An extraction tube, the extraction tube comprising a bottle body, a detachable upper cover is provided at the opening of the bottle body, an inner plug is provided between the upper cover and the bottle body, the inner plug is arranged in a funnel shape, and the thin mouth end of the inner plug is in a pressure valve shape and is located on the side of the upper cover, so that when the bottle body is inverted, the sample liquid will not drip freely. A sample extraction liquid capable of simultaneously processing multiple antigens is provided in the bottle body. The sample extraction liquid comprises a buffer solution and a lysis surfactant. Among them, the formulation ratio of the buffer solution to the surfactant is PBS at 0.01 mol / L and SDS solution with a concentration of 0.1%, and the pH is 7.

4. A liquid dropping port is provided at the top of the upper cover for dropping the sample, and a detachable top cover is provided at the liquid dropping port. The extraction tube is used for processing the sample and dropping the sample onto the test strip; Test strips, each test strip comprising a sample pad, a conjugate pad, a chromatographic membrane and an absorbent pad arranged on a PVC bottom plate. A test line and a quality control line are provided on the chromatographic membrane. The test line is coated with an antibody for detecting the virus, and the quality control line is coated with a goat anti-mouse IgG antibody. The multiple test strips are fixedly arranged on the base.

2. The kit for the combined detection of multiple virus antigens according to claim 1, characterized in that, A conjugate pad is arranged on one side of the chromatographic membrane, and different virus antibodies labeled with colloidal gold are distributed on the conjugate pads of different test strips. Among them, the virus antibodies are novel coronavirus antibodies, influenza A / B virus antibodies, and respiratory syncytial virus antibodies.

3. The kit for the combined detection of multiple virus antigens according to claim 2, characterized in that, An absorbent pad is arranged on the other side of the chromatographic membrane, and the sample pad is arranged on the side of the conjugate pad away from the absorbent pad.

4. The kit for the combined detection of multiple virus antigens according to claim 3, characterized in that, The sample pad is a first glass fiber membrane dried after being soaked in a preset sample pad treatment liquid. The sample pad treatment liquid comprises a buffer solution, a hydrophilic surfactant, and casein. Among them, the formulation ratio of the buffer solution, the surfactant, and the casein is PBS at 0.01 mol / L, Tween-20 solution with a concentration of 0.4%, and casein with a concentration of 0.5%, and the pH is 7.

4.

5. The kit for the combined detection of multiple virus antigens according to claim 3, characterized in that, The conjugate pad is a second glass fiber membrane dried after being soaked in a preset conjugate pad pretreatment liquid. The conjugate pad pretreatment liquid comprises a buffer agent, a surfactant, casein, and a preservative. Among them, the formulation of the conjugate pad pretreatment liquid is PB at 0.01 mol / L, casein at a concentration of 0.3%, Tween-20 at a concentration of 0.5%, and PC300 preservative at a concentration of 0.01%, and the pH value is 7.

4.

6. The kit for the combined detection of multiple virus antigens according to claim 1, characterized in that, The virus antibodies coated on the test lines of different chromatographic membranes are different, and a quality control line coated with the same goat anti-mouse IgG antibody is also provided on each chromatographic membrane.

7. The kit for the combined detection of multiple virus antigens according to claim 5, characterized in that, The chromatographic membrane includes an NC membrane. The test line is drawn on the NC membrane with a coating buffer containing a virus antibody, and the control line is drawn on the NC membrane with a coating buffer containing a control antibody. The coating buffer includes a buffer, trehalose, and a preservative. Among them, the formulation ratio of the buffer, the trehalose, and the preservative is 0.01 mol / L PB buffer, 1% trehalose, and 0.01% PC300 preservative.

8. The application method of the kit for the combined detection of multiple virus antigens according to any one of claims 1 to 7, characterized in that, The method is for non-diagnostic purposes. The application method of the multiple virus antigen combined detection kit includes: Placing the collected sample in an extraction tube; Squeezing the bottle body of the extraction tube multiple times and standing for a preset period of time to obtain a sample solution. Among them, the extraction tube contains a sample extraction solution that can simultaneously process multiple antigens, and the sample extraction solution can simultaneously lyse multiple viruses to expose the nucleoproteins of multiple viruses; Dropping the sample solution on multiple preset test strips and waiting for the antigens in the sample solution to react with the antibodies on the test strips; After the antigens in the sample solution react with the antibodies on the test strips, based on the control line and the test line set on the test strip, a test result is obtained.

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