阅读说明：本技术 金刚乙胺半抗原、金刚乙胺完全抗原及其制备方法与试纸条 (Rimantadine hapten, rimantadine complete antigen, preparation method thereof and test strip ) 是由 蒋永青 张伟涛 王金苹 薛华平 于 2020-05-29 设计创作，主要内容包括：本发明公开了一种金刚乙胺半抗原、完全抗原及其制备方法与试纸条,该金刚乙胺完全抗原中,其金刚乙胺分子的抗原决定簇能充分暴露,且具有较完整的空间构象。因此,本发明提供的金刚乙胺完全抗原具有较好的免疫原性,可用于免疫分析。(The invention discloses a rimantadine hapten, a complete antigen, a preparation method thereof and a test strip. Therefore, the rimantadine complete antigen provided by the invention has better immunogenicity and can be used for immunoassay.)

1. A rimantadine hapten, characterized by the structural formula:

2. a preparation method of rimantadine hapten is characterized by comprising the following steps:

providing rimantadine of the formula

Dissolving rimantadine and succinic anhydride in pyridine according to a molar ratio of 1: 1-1: 2 to obtain a reaction solution, reacting the reaction solution at 80-100 ℃ for 2-3 hours, and separating and purifying the reaction solution to obtain the rimantadine hapten.

3. The rimantadine complete antigen is characterized by having a structural formula

In the formula, R is carrier protein, and n is a natural number of 5-20.

4. A preparation method of a rimantadine complete antigen is characterized by comprising the following steps:

dissolving rimantadine hapten in N, N-dimethylformamide solution, and then adding 1-ethyl-carbodiimide hydrochloride and N-hydroxysuccinimide, or adding 3-dimethylaminopropyl-carbodiimide hydrochloride and N-hydroxysuccinimide to obtain first reaction liquid;

stirring and reacting for 12-16 hours at room temperature to obtain a second reaction solution;

preparing a phosphate buffer solution of the carrier protein;

dropwise adding the second reaction solution into the phosphate buffer solution of the carrier protein, stirring at room temperature for reacting overnight to obtain a mixed solution containing rimantadine complete antigen, dialyzing the mixed solution containing rimantadine complete antigen, and centrifuging to obtain a supernatant solution containing the rimantadine complete antigen, wherein the structural formula of the rimantadine complete antigen is shown in the specification

In the formula, R is carrier protein, and n is a natural number of 5-20.

5. Rimantadine complete antigen according to claim 3, characterized in that the carrier protein is any of globulin, bovine serum albumin, chicken ovalbumin, keyhole limpet hemocyanin, rabbit serum albumin, human serum albumin, thyroglobulin, fibrinogen, rabbit gamma globulin and chicken gamma globulin.

6. A colloidal gold immunochromatographic test strip for detecting rimantadine is characterized by comprising a back plate, a sample pad, a gold label pad, a reaction pad and a water absorption pad; the sample pad, the gold label pad, the reaction pad and the water absorption pad are sequentially arranged on the back plate; the reaction pad is provided with a detection line and a quality control line, the detection line is coated with a rimantadine complete antigen, the quality control line is coated with an anti-rimantadine antibody gold-labeled compound, and the gold-labeled pad is sprayed with the rimantadine antibody gold-labeled compound.

7. The colloidal gold immunochromatographic strip for detecting rimantadine according to claim 6, wherein the rimantadine antibody gold-labeled complex comprises rimantadine antibody and colloidal gold nanoparticles, and the rimantadine antibody gold-labeled complex has the ability to recognize and specifically bind to rimantadine.

8. The colloidal gold immunochromatographic strip for detecting rimantadine according to claim 7, wherein the rimantadine antibody-gold labeled complex is formed by electrostatically binding the rimantadine antibody and the colloidal gold nanoparticles.

9. The colloidal gold immunochromatographic test strip for detecting rimantadine according to claim 7, wherein the rimantadine antibody is a monoclonal antibody or a polyclonal antibody.

10. The colloidal gold immunochromatographic test strip for detecting rimantadine according to claim 9, wherein the polyclonal antibody is any one of murine, equine, ovine, rabbit or guinea pig origin, and the monoclonal antibody is murine or rabbit origin.

Technical Field

The invention relates to the field of biomedical materials, in particular to a rimantadine hapten, a rimantadine complete antigen, a preparation method thereof and a test strip.

Background

The immunoassay is a trace analysis method based on specific recognition and reversible binding reaction between an antigen and an antibody, and is suitable for detection of macromolecular compounds (such as proteins, nucleic acids and bacteria) and measurement of small molecular compounds (such as hormones and drugs). The key to establishing an immunoassay method for small molecule compounds is the ability to prepare antibodies with high affinity and high selectivity for small molecule compounds. Since most small molecule compounds have a molecular weight of less than 1000, they are not immunogenic, i.e., lack T cell epitopes and cannot directly induce the production of specific antibodies in animal organisms and the like. However, the small molecule compound can be combined with the macromolecular carrier by a proper chemical modification method to generate a small molecule compound-macromolecular carrier conjugate, the conjugate is a complete antigen, and the small molecule compound-macromolecular carrier conjugate can indirectly induce the proliferation and differentiation of B cells by means of T cell epitopes to generate specific antibodies.

Rimantadine (MLT) is a small molecule substance with the chemical name of N-acetyl-5-methoxytryptamine, the molecular weight of which is only 232.28D, and it is not immunogenic, and it is used for immunization, and it must be combined with protein macromolecular carrier first to form rimantadine-protein carrier coupled complete antigen. However, the preparation of rimantadine-protein carrier conjugated complete antigen does not affect the spatial conformation of rimantadine, and requires the maintenance of the availability of antigenic determinants of rimantadine (e.g., 5-methoxy, N-alkyl side chains) to obtain a high potency, high specificity rimantadine antibody. Therefore, how to prepare a complete antigen with good immunogenicity is a difficult point in establishing an immunoassay method for measuring rimantadine.

Disclosure of Invention

The invention provides a rimantadine hapten, a rimantadine complete antigen, a preparation method thereof and a test strip, and aims to solve the problems.

According to a first aspect of embodiments of the present application, there is provided a rimantadine hapten of the formula:

according to a second aspect of the embodiments of the present application, there is provided a method for preparing a rimantadine hapten, comprising the steps of:

providing rimantadine of the formula

Dissolving rimantadine and succinic anhydride in pyridine according to a molar ratio of 1: 1-1: 2 to obtain a reaction solution, reacting the reaction solution at 80-100 ℃ for 2-3 hours, and separating and purifying the reaction solution to obtain the rimantadine hapten.

According to a third aspect of the embodiments of the present application, there is provided a rimantadine complete antigen having the structural formula

In the formula, R is carrier protein, and n is a natural number of 5-20.

According to a fourth aspect of the embodiments of the present application, there is provided a method for preparing a rimantadine complete antigen, comprising the steps of:

dissolving rimantadine hapten in N, N-dimethylformamide solution, and then adding 1-ethyl-carbodiimide hydrochloride and N-hydroxysuccinimide, or adding 3-dimethylaminopropyl-carbodiimide hydrochloride and N-hydroxysuccinimide to obtain first reaction liquid;

stirring and reacting for 12-16 hours at room temperature to obtain a second reaction solution;

preparing a phosphate buffer solution of the carrier protein;

dropwise adding the second reaction solution into the phosphate buffer solution of the carrier protein, stirring at room temperature for reacting overnight to obtain a mixed solution containing rimantadine complete antigen, dialyzing the mixed solution containing rimantadine complete antigen, and centrifuging to obtain a supernatant solution containing the rimantadine complete antigen, wherein the structural formula of the rimantadine complete antigen is shown in the specification

In the formula, R is carrier protein, and n is a natural number of 5-20.

According to a fifth aspect of the embodiments of the present application, there is provided a colloidal gold immunochromatographic test strip for detecting rimantadine, comprising a back plate, a sample pad, a gold-labeled pad, a reaction pad, and a water-absorbing pad; the sample pad, the gold label pad, the reaction pad and the water absorption pad are sequentially arranged on the back plate; the reaction pad is provided with a detection line and a quality control line, the detection line is coated with a rimantadine complete antigen, the quality control line is coated with an anti-rimantadine antibody gold-labeled compound, and the gold-labeled pad is sprayed with the rimantadine antibody gold-labeled compound.

The technical scheme provided by the embodiment of the application can have the following beneficial effects:

the rimantadine complete antigen provided by the invention comprises rimantadine molecules and carrier protein, wherein the rimantadine molecules and the carrier protein are connected through a connecting arm, and in the rimantadine complete antigen, an antigenic determinant of the rimantadine molecule can be fully exposed and has a relatively complete spatial conformation, so that the rimantadine complete antigen provided by the invention has relatively good immunogenicity.

The colloidal gold immunochromatographic kit for detecting rimantadine provided by the invention can be used for immunodetection of rimantadine, the detection sensitivity is greatly improved, all positive samples of rimantadine can be rapidly screened out through the colloidal gold immunochromatographic kit, and a sample larger than 1.8ng/ml can be detected.

The colloidal gold immunochromatographic kit for detecting rimantadine provided by the invention has short detection time; no special instrument is needed; the operation is simple and convenient, the operator does not need to be trained, and the detection cost is low; in a word, the detection kit is simple, economical and convenient to use.

The preparation method of the colloidal gold immunochromatographic kit for detecting rimantadine provided by the invention is simple and easy to implement, the adopted test paper is conventional test paper, the adopted antibody can be prepared according to the method provided by the invention, the large-scale production can be realized, and the production cost is low.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.

Fig. 1 is a schematic structural diagram of a colloidal gold immunochromatographic test strip for detecting rimantadine in an embodiment of the present invention.

Description of reference numerals:

10. a back plate; 20. a sample pad; 30. a gold label pad; 40. a reaction pad; 41. detecting lines; 42. a quality control line; 50. an absorbent pad.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

It is also to be understood that the terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used in the specification of the present invention and the appended claims, the singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

It should be further understood that the term "and/or" as used in this specification and the appended claims refers to and includes any and all possible combinations of one or more of the associated listed items.

The reagents and biomaterials in the examples described below are commercially available without specific reference.

The invention discloses a rimantadine hapten with a structural formula

The invention discloses a preparation method of rimantadine hapten, which comprises the steps of S101-S102.

S101, providing rimantadine with a structural formula

S102, dissolving the rimantadine and succinic anhydride in pyridine according to a molar ratio of 1: 1-1: 2 to obtain a reaction solution, reacting the reaction solution at 80-100 ℃ for 2-3 hours, and separating and purifying the reaction solution to obtain the rimantadine hapten.

In an alternative embodiment, the invention discloses a rimantadine complete antigen with the structural formula

In the formula, R is carrier protein, and n is a natural number of 5-20.

The preparation method of the rimantadine complete antigen includes steps S201-S202.

S201, dissolving rimantadine hapten in N, N-dimethylformamide solution, and then adding 1-ethyl-carbonyldiimine hydrochloride and N-hydroxysuccinimide, or adding 3-dimethylaminopropyl-carbonyldiimine hydrochloride and N-hydroxysuccinimide to obtain a first reaction liquid;

s202, stirring and reacting at room temperature for 12-16 hours to obtain a second reaction solution.

Specifically, in the first reaction solution, the concentration of rimantadine hapten was 0.05

The molar concentrations of mol/L, 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine hydrochloride and N-hydroxysuccinimide are respectively 1.2 times of the molar concentration of the rimantadine hapten;

s203, preparing a phosphate buffer solution of carrier protein.

Specifically, the molar concentration of the carrier protein is 0.001mol/L, and the molar concentration of the phosphate is 0.01 mol/L.

Dropwise adding the second reaction solution into the phosphate buffer solution of the carrier protein, stirring at room temperature for reacting overnight to obtain a mixed solution containing rimantadine complete antigen, dialyzing the mixed solution containing rimantadine complete antigen, and centrifuging to obtain a supernatant solution containing the rimantadine complete antigen, wherein the structural formula of the rimantadine complete antigen is shown in the specification

In the formula, R is carrier protein, and n is a natural number of 5-20.

In an alternative embodiment, the carrier protein is any one of globulin, bovine serum albumin, chicken ovalbumin, keyhole limpet hemocyanin, rabbit serum albumin, human serum albumin, thyroglobulin, fibrinogen, rabbit gamma globulin, and chicken gamma globulin.

The invention discloses a colloidal gold immunochromatographic test strip for detecting rimantadine, which comprises a back plate, a sample pad, a gold label pad, a reaction pad and a water absorption pad, wherein the sample pad, the gold label pad, the reaction pad and the water absorption pad are sequentially arranged on the back plate, the sample pad and the water absorption pad are respectively arranged at two ends of the back plate, the reaction pad is arranged in the middle of the back plate, the lower surface of the end part of the gold label pad close to the reaction pad is attached to the upper surface of the reaction pad, and the upper surface of the end part of the gold label pad close to the sample pad is attached to the lower surface of the sample pad. The upper surface of one end of the reaction pad, which is far away from the gold mark pad, is attached to the upper surface of the water absorption pad, so that the liquid to be detected is fully overflowed onto the reaction pad through the structure and is finally absorbed by the water absorption pad. The reaction pad is provided with a detection line and a quality control line which are arranged in parallel, the detection line is coated with a rimantadine complete antigen, the quality control line is coated with an anti-rimantadine antibody gold-labeled compound, and the gold-labeled pad is sprayed with the rimantadine antibody gold-labeled compound.

The invention also provides a preparation method of the colloidal gold immunochromatographic test strip for detecting rimantadine, which comprises the following steps S301-S303.

S301, preparing a rimantadine antibody, colloidal gold and a gold-labeled pad.

Specifically, the rimantadine antibody is prepared by taking a rimantadine complete antigen as an immunogen and adopting a conventional method. The colloidal gold is prepared into the colloidal gold with the particle size of 25-40 nm by adopting a trisodium citrate reduction method.

The preparation method of the gold-labeled pad comprises the following steps of dipping the glass fiber treated by the treatment solution, drying, spraying the rimantadine antibody gold-labeled compound on the pretreated glass fiber in an amount of 0.5-4 mu l/cm, and drying at 25-30 ℃ to obtain the gold-labeled pad (3) which is placed in an environment of 2-8 ℃ for later use.

When the rimantadine antibody gold-labeled compound is prepared, the pH value of colloidal gold is adjusted to 7.0-9.0, the rimantadine antibody is slowly added according to the dosage of adding 4-25 mu g of protein into each milliliter of colloidal gold solution under the stirring condition, the mixture is kept stand for 10-30 min, bovine serum albumin BSA is added to the final concentration of 0.5-5%, the mixture is kept stand for 10-30 min, the mixture is centrifuged, the supernatant is discarded, the precipitate is washed for 2-3 times by using a washing solution, the precipitate is re-suspended by using a ten-tenth initial volume of re-suspension at the last time, and the re-suspension is kept at 4 ℃.

And S302, setting a detection line and a quality control line.

Specifically, the detection line is set by diluting the rimantadine complete antigen with a coating buffer solution to a concentration of 1.0-1.5 mg/mL; and taking the reaction pad, and scratching the rimantadine complete antigen on the reaction pad by using a film scratching instrument with the dosage of 1-10 mu l/cm to form a detection line.

Setting a quality control line, diluting the goat anti-mouse IgG to the concentration of 0.8-1.5 mg/mL by using a coating buffer solution, then scratching the goat anti-mouse IgG on a reaction pad by using a film scratching instrument at the dosage of 1-10 mu l/cm to form the quality control line, and then placing the reaction pad at 37 ℃ for drying for later use.

The distance between the detection line and the quality control line is set to be within the range of 5-8 mm, the thickness of the two lines of spraying lines is uniform, and the reaction pad is packaged for later use after being dried at 37 ℃; the reaction pad is made of NC membrane, which can be any commercially available nitrocellulose membrane, preferably S & S AE99, whatman 8 μ M, millipore M135, sartorius CN 140.

The coating buffer may be borate, carbonate, phosphate, Tris-HCl or Tris-phosphate, acetate, barbiturate, etc., and the purpose of the buffer is to provide a pH and ionic strength to coat and immobilize the protein on the NC membrane, wherein the pH of the buffer is generally in the range of about 6-9.5, preferably in the range of neutral buffer of 6.5-7.5, and most preferably in the range of 7.0-7.4. Preferably a phosphate. Adding 8.0g of NaCI, 40.2 g of KH2PO40, 4.12H2O2.9g of Na2HPO4 and 0.2g of KCI into a quantitative container in sequence, adding a proper amount of distilled water for dissolving, then fixing the volume to 1000mL, adjusting the pH value to 7.4, sterilizing 112Kpa for 20min under high pressure, cooling, and storing in a refrigerator at 4 ℃ for later use.

S303, assembling the colloidal gold immunochromatographic test strip for detecting rimantadine.

Specifically, get backplate, sample pad, absorb water the pad, the gold mark pad after handling and the reaction pad after handling and assemble, obtain the colloidal gold immunochromatographic assay kit that detects benzoic acid, wherein sample pad, gold mark pad, reaction pad and absorb water the pad set gradually in on the backplate, sample pad and absorb water the pad and set up respectively in two tip of backplate, the reaction pad sets up in the middle part of backplate, the lower surface that the tip that is close to the reaction pad of gold mark pad is laminated with the upper surface of reaction pad, the upper surface that is close to the tip of sample pad on the gold mark pad is laminated with the lower surface of sample pad. The upper surface of one end of the reaction pad, which is far away from the gold mark pad, is attached to the upper surface of the water absorption pad, so that the liquid to be detected is fully overflowed onto the reaction pad through the structure and is finally absorbed by the water absorption pad. The detection line is coated with a rimantadine complete antigen, the quality control line is coated with an anti-rimantadine antibody gold-labeled compound, and the gold-labeled pad is sprayed with the rimantadine antibody gold-labeled compound.

In some optional embodiments, the test strip may be mounted in a housing, the reaction pad is exposed to allow a user to observe the reaction of the reaction pad externally, and the housing corresponding to the sample pad is also provided with an opening to allow the user to insert the sample onto the sample pad through the opening.

In this embodiment, the material of the back plate is a PVC plate, the material of the sample pad is glass fiber, and the material of the absorbent pad is absorbent paper. When assembling, the indoor temperature should be controlled at 25-37 deg.C and humidity 20-30%.

In an alternative embodiment, the rimantadine antibody gold-labeled complex comprises rimantadine antibody and colloidal gold nanoparticles, the rimantadine antibody gold-labeled complex having the ability to recognize and specifically bind to rimantadine. The rimantadine antibody is a monoclonal antibody or a polyclonal antibody. The rimantadine antibody gold-labeled compound is formed by electrostatically combining a rimantadine antibody and colloidal gold nanoparticles.

In an alternative embodiment, the rimantadine antibody is a monoclonal antibody and the method of preparation comprises steps S301-S304.

S301, animal immunization.

The conjugate MLT-BSA is used as an immunizing antigen to immunize Balb/c mice, and the weight is 100 mug/kg. Mixing immunogen and equivalent Freund complete adjuvant to prepare emulsifier during first immunization, injecting subcutaneously at neck and back at multiple points, taking the same dose of immunogen and equivalent Freund incomplete adjuvant at intervals of 2 weeks, mixing and emulsifying, continuing third immunization, taking blood at tail part after one week of third immunization to detect serum titer and inhibition, performing abdominal cavity impact immunization once when the requirement is detected, and taking splenocytes after 3 days.

S302, cell fusion and cloning.

Taking splenocytes of the immune Balb/c mice, and carrying out the following steps: 1 ratio was fused with SP2/0 myeloma cells, and positive wells were screened by measuring cell supernatants using an indirect competitive ELISA. Cloning the positive hole by using a limiting dilution method until obtaining the hybridoma cell strain which stably secretes the monoclonal antibody.

S303, freezing and restoring cells.

Preparing hybridoma cells in logarithmic growth phase into 5 xl 06 cell suspension with freezing medium, subpackaging in freezing tube, and storing in liquid nitrogen for a long time. Taking out the frozen tube during recovery, immediately putting the tube into a water bath at 37 ℃ for fast melting, centrifuging to remove frozen liquid, and transferring the tube into a culture bottle for culture.

S304, preparing and purifying the monoclonal antibody.

By adopting an in vivo induction method, 0.5ml of Freund's incomplete adjuvant is injected into the abdominal cavity of a Balb/c mouse aged for 8 weeks, 5 xl 06 hybridoma cells are injected into the abdominal cavity after 7 days, and ascites is collected after 7 days. Purifying ascites with affinity chromatographic column, measuring protein concentration with ultraviolet spectrophotometer, and storing at-20 deg.c. The titer of the purified antibody was measured by ELISA (1:128000), and the results showed that the purified antibody showed high specificity and sensitivity to rimantadine.

In an alternative embodiment, the rimantadine antibody is a polyclonal antibody and the method of preparation comprises steps S401-S402.

S401, animal immunization.

The conjugate MLT-BSA provided by the embodiment of the invention is used as an immunizing antigen to immunize Balb/c mice of 8 weeks old, and the weight is 1 mg/kg. Mixing the immunogen and equivalent Freund's complete adjuvant to prepare an emulsifier during first immunization, performing subcutaneous multi-point injection on the neck and back, taking the same dose of immunogen and equivalent Freund's incomplete adjuvant at intervals of 2 weeks, mixing and emulsifying, and performing booster immunization once for 5 times.

S402, collecting antiserum and purifying polyclonal antibody.

Blood is collected 7 days after the last immunization, the titer of serum antibodies is measured, abdominal artery is collected with whole blood to collect serum, purified polyclonal antibodies are obtained by affinity chromatography column separation, the titer (1:10000) of the purified antibodies is measured by ELISA method, and the purified antibodies show higher specificity and sensitivity to rimantadine.

In an alternative embodiment, the method for preparing colloidal gold includes steps S501-S503.

S501, dissolving 1g of chloroauric acid (HauCL4) in 100g of ultrapure water to prepare a 1% chloroauric acid solution 4C, and placing for later use;

s502, dissolving 1g of trisodium citrate dihydrate into 100g of ultrapure water to prepare 1% trisodium citrate dihydrate solution 4C, and placing for later use;

s503, adding 100ml of ultrapure water into a clean triangular flask, placing the flask on a heated magnetic stirrer, stirring and heating the flask to boiling, then adding 1ml of 1% chloroauric acid, quickly adding 1.7ml of 1% trisodium citrate dihydrate solution after the flask is boiled, continuing heating and stirring for 15 minutes after the color is changed into wine red, and then turning off heating and stirring. Naturally cooling the prepared colloidal gold for later use; the particle size of the prepared colloidal gold is 20-40 nm, and the maximum absorption peak is 521 nm.

In an alternative embodiment, the preparation method of the rimantadine antibody gold-labeled complex comprises steps S601 to S602:

s601, searching the most suitable protein concentration marked by colloidal gold

Taking 8 polystyrene micro-reaction tubes, adding 30ul of triple distilled water into each hole, adding 30ul of the rimantadine antibody prepared in the example IV into the first hole, diluting the mixture in a multiple ratio to the last hole, adding 125ul of Ph8.0 colloidal gold solution into each hole, standing the mixture at room temperature for 15min, adding 125ul of 10% NaCl solution into each hole, observing the change of each hole, and taking the hole without color change as the minimum protein concentration (5ug/ml-10 ug/ml).

S602, adjusting the pH value of colloidal gold to 8.0, slowly adding the rimantadine antibody according to the dosage of adding 6 mu g of protein into each milliliter of colloidal gold solution under the stirring condition, standing for 30min, then adding bovine serum albumin BSA to the final concentration of 1%, standing for 30min, centrifuging for 30min at 4 ℃ under 12000 rpm, discarding the supernatant, washing the precipitate for 2 times by using a washing solution, re-suspending the precipitate by using a tenth of the initial volume of a resuspension solution at the last time to obtain an rimantadine antibody gold-labeled compound, and standing for later use at 4 ℃.

The resuspension solution, namely the preservation solution, is borate, carbonate, phosphate, Tris-HCl or Tris-phosphate, acetate, barbital and the like containing 0.2-1% of macromolecular protein, the macromolecular protein can be bovine serum albumin, PEG20000, casein and the like, and the buffer solution is preferably a phosphate buffer solution, more preferably 0.5% BSA phosphate buffer solution with the pH value of 7.4.

In addition, the preparation method of the colloidal gold immunochromatographic kit for detecting rimantadine provided in this embodiment is only one of the ways of applying the rimantadine complete antigen, the rimantadine antibody gold-labeled complex, and the antibody of the anti-rimantadine antibody gold-labeled complex provided by the present invention, and other applications, particularly, immunoassay application of rimantadine, should be considered as the protection scope of the embodiment of the present invention; in addition, the colloidal gold immunochromatographic kit for detecting rimantadine provided by the invention can be packaged, and comprises the following steps: and (3) assembling 1 part of the cut test paper in the prepared test paper card, enabling the sample adding window to correspond to the sample pad of the test paper, and enabling the result display window to correspond to the detection area and the control area. And packaging the dried powder, a package of drying agent, a specification and a sample injector in an outer packaging bag, and storing the packaged powder in a dark place at 4-25 ℃.

While the invention has been described with reference to specific embodiments, the invention is not limited thereto, and various equivalent modifications and substitutions can be easily made by those skilled in the art within the technical scope of the invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.