Etomidate complete antigen, and preparation method therefor and use thereof

Abstract

Provided are an etomidate complete antigen, and a preparation method therefor and the use thereof. The preparation method comprises: preparing an etomidate hapten derivative solution from an etomidate hapten; dissolving a carrier protein to prepare a carrier protein solution; and mixing the etomidate hapten derivative solution with the carrier protein solution to perform a coupling reaction to obtain an etomidate complete antigen. The provided technical solution has the characteristics of simplicity, rapidity, high specificity, broad applicability and high sensitivity; and has good application prospects in the rapid and effective detection of etomidate.

Description

Etoimidazole complete antigen and preparation method and application thereof TECHNICAL FIELD

[0001] The present application relates to the field of biological detection, in particular to a kind of etomidate complete antigen and preparation method and application thereof. BACKGROUND

[0002] Etoimidazole is one of the commonly used drugs for anesthesia induction (molecular formula C 14 H 16 N2O2), and has been used in clinical application for 30 years. It is generally used intravenously, and has rapid but short-acting, fast sleep and fast recovery. It has a strong inhibitory effect on the central nervous system. Under the use of conventional dose, etomidate has the characteristics of slight influence on cardiovascular function and respiratory system, and fast in vivo metabolism and clearance. It is used as an intravenous general anesthetic induction drug or anesthetic adjuvant in clinical application, and is often used in combination with other anesthetic drugs.

[0003] Currently, the detection of etomidate mainly relies on high performance liquid chromatography (HPLC), gas chromatography (GC), thin layer chromatography (TLC), mass spectrometry (MS) and the like. However, the chromatographic detection has the disadvantages of expensive instrument, time-consuming detection and the need for professional technical personnel to operate, and cannot meet the requirements of modern detection for rapidness and accuracy. In addition, immunological detection methods such as ELISA and immunochromatographic test paper technology have also developed rapidly in recent years. Enzyme-linked immunosorbent assay has the characteristics of simple sample pretreatment, simple operation, rapidness, sensitivity and high throughput, and is praised as the most competitive and challenging rapid detection technology in the 21st century, and has a broad application prospect in the field of drug detection. However, for the immunological analysis method, the effect of the antibody as the core raw material is greatly dependent on the antigen structure that causes the immune response of the corresponding animal. The patent application with publication number CN116354888 discloses a kind of etomidate artificial hapten and artificial antigen, which cannot meet the requirements of sensitive and accurate detection of etomidate. And there are few etomidate detection reagent products on the market, and the sensitivity is low. Therefore, it is of great significance to provide a method for preparing a high-specificity antibody that can rapidly, sensitively and accurately detect etomidate with stable antigen structure, simple operation and wide application. SUMMARY

[0004] In view of the above-mentioned deficiencies in the detection of etomidate at present, the present application provides a kind of etomidate complete antigen, which can achieve the effects of stable antigen structure, simple operation and wide application.

[0005] To achieve the above-mentioned purpose, the embodiments of the present application adopt the following technical solutions:

[0006] A method for preparing a complete etomidate antigen, comprising the following steps:

[0007] A solution of etomidate hapten derivative is prepared by the etomidate hapten;

[0008] A carrier protein solution is prepared by dissolving a carrier protein;

[0009] The etomidate complete antigen is prepared by mixing the solution of etomidate hapten derivative and the carrier protein solution for coupling reaction.

[0010] According to an aspect of the present application, the etomidate hapten is prepared by reacting etomidate with a methanol solution under alkaline conditions to obtain an intermediate, and then hydrolyzing and extracting the intermediate.

[0011] According to an aspect of the present application, the solution of etomidate hapten derivative is prepared by dissolving the etomidate hapten in an organic solvent, adding N-hydroxysuccinimide and carbodiimide, stirring and reacting to obtain the solution of etomidate hapten derivative.

[0012] According to an aspect of the present application, the organic solvent is DMF, and the ratio of the etomidate hapten, N-hydroxysuccinimide and carbodiimide is 5:4:5.

[0013] According to an aspect of the present application, the carrier protein solution is prepared by dissolving a carrier protein in a buffer to obtain the carrier protein solution, wherein the carrier protein is selected from bovine serum albumin or hemocyanin, and the buffer is PBS buffer.

[0014] According to an aspect of the present application, the mass ratio of the etomidate hapten derivative to the carrier protein is 1:2-20.

[0015] According to an aspect of the present application, in the step of mixing the solution of etomidate hapten derivative and the carrier protein solution for coupling reaction to obtain the etomidate complete antigen, low-temperature dialysis is performed after the coupling reaction, and the low-temperature dialysis time is 1-3 days.

[0016] According to an aspect of the present application, the present application further provides an etomidate complete antigen prepared by the above method.

[0017] According to an aspect of the present application, the present application further provides any of the following applications of the etomidate complete antigen:

[0018] 1) for detecting etomidate and its homologues;

[0019] 2) for preparing an immunochromatographic reagent strip of etomidate.

[0020] 3) for preparing etomidate test paper or kit.

[0021] According to one aspect of the present application, the present application also provides a reagent strip, which comprises a substrate, a sample adding part, a detection part and a liquid absorbing part, the sample adding part adsorbs a colloidal gold labeled etomidate clone antibody prepared by etomidate complete antigen, the detection part is fixed on the substrate, a quality control band and a detection band are arranged in the middle of the detection part, and the liquid absorbing part and the sample adding part are fixed at both ends of the detection part in a partially overlapping manner, wherein the detection band is coated with the above-mentioned etomidate complete antigen, and the quality control band is coated with goat anti-rabbit IgG.

[0022] According to one aspect of the present application, the substrate is a PVC lining board, and the detection part is a nitrocellulose membrane.

[0023] According to one aspect of the present application, the present application also provides a detection kit, which comprises the above-mentioned etomidate immunochromatography reagent strip and etomidate detection analysis liquid matched with the above-mentioned etomidate immunochromatography reagent strip.

[0024] The present application has the following advantages: through the above technical solution, the etomidate hapten provided by the present application retains the characteristic structure of etomidate to the greatest extent, and further utilizes the complete antigen obtained by coupling the etomidate hapten with a carrier protein as an immunogen to immunize New Zealand white rabbits, so that the etomidate monoclonal antibody is prepared.

[0025] The anti-etomidate antibody prepared by the present application has good affinity, sensitivity and specificity, and the titer of the immune serum obtained by immunization is as high as 10 5 The above can be used for rapid and accurate immunodetection and immunological analysis of etomidate.

[0026] The present application has the characteristics of simplicity, rapidness, strong specificity, wide application range and high sensitivity, and has good application prospect and broad development space in the rapid and effective detection of etomidate. BRIEF DESCRIPTION OF DRAWINGS

[0027] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed in the embodiments will be briefly introduced as follows. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative labor.

[0028] Fig. 1 is a structural schematic view of an etomidate immunochromatography reagent strip according to the present application;

[0029] Fig. 2 is a partial enlarged view of position A in Fig. 1.

[0030] 1, absorbent stick; 2, sample pad; 3, reagent pad; 4, nitrocellulose membrane; 5, water absorption pad; 6, backing plate; 7, quality control line; 8, detection line; 9, sample addition position; 10, liquid chromatography direction. DETAILED DESCRIPTION

[0031] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application. Embodiment 1

[0032] A preparation method of a complete etomidate antigen, the preparation method of the complete etomidate antigen comprising the following steps:

[0033] Step S1: obtaining an etomidate hapten derivative solution by preparing an etomidate hapten;

[0034] The step of obtaining the etomidate hapten derivative solution by preparing an etomidate hapten specifically comprises the following steps:

[0035] Step S11: synthesis of an etomidate artificial hapten;

[0036] Dissolve etomidate in a KOH methanol solution, stir and react at room temperature, evaporate the solvent under reduced pressure after complete reaction, dissolve the residue in water, extract with diethyl ether, remove the organic phase, place the water phase in an ice water bath, acidify with hydrochloric acid, adjust the pH value, precipitate the solid, filter and dry to obtain the hydrolysis product as the etomidate hapten.

[0037] Step S12: dissolve the etomidate hapten in an organic solvent, add N-hydroxysuccinimide and carbodiimide, stir and react to prepare an etomidate hapten derivative solution;

[0038] The organic solvent is DMF, and the ratio of the etomidate hapten, N-hydroxysuccinimide and carbodiimide is 5:4:5.

[0039] Step S2: dissolving a carrier protein to prepare a carrier protein solution;

[0040] Dissolve the carrier protein in a buffer solution to prepare the carrier protein solution, the buffer solution being a PBS buffer solution comprising the following components: NaCl, NaH2PO4, Na2HPO4.12H2O and H2O.

[0041] Step S3: mixing the etomidate hapten derivative solution with the carrier protein solution to carry out a coupling reaction to obtain an etomidate complete antigen.

[0042] The carrier protein solution was stirred, and the etomidate hapten derivative solution was added dropwise into the carrier protein solution, and the reaction was continued, and then low-temperature dialysis was carried out in a PBS dialysis solution to obtain the etomidate complete antigen. Example 2

[0043] Etidomide was dissolved in 10% KOH methanol solution, stirred at room temperature, and the reaction was tracked by TLC. After complete reaction, the solvent was evaporated under reduced pressure, the residue was dissolved in water, and extracted twice with diethyl ether, the organic phase was removed, and the water phase was placed in an ice water bath and acidified with 1 mol / L hydrochloric acid to adjust the pH value to 6-7. The precipitated solid was filtered and dried to obtain the hydrolysis product as the etomidate hapten.

[0044] 10 mg of etomidate hapten was dissolved in 0.5 mL of DMF (N,N-dimethylformamide), 8 mg of NHS (N-hydroxysuccinimide) and 10 mg of EDC (carbonyldiimidazole) were added, and the etomidate hapten derivative solution was obtained after stirring until the reaction was complete. 50 mg of BSA (bovine serum albumin) was dissolved in 2 mL of PBS (pH = 7.4), and the etomidate hapten derivative solution was added dropwise while stirring, and the reaction was continued for 12 hours. The preparation method of the PBS solution: 8.5 g of NaCl, 0.3 g of NaH2PO4, and 2.9 g of Na2HPO4.12H2O were added to water to make up to 1 L, and the pH value was adjusted to 7.4, and it was stored at 4°C for standby. Finally, low-temperature dialysis was carried out in a PBS dialysis solution for 3 days. It was stored for detection. The mass ratio of the etomidate hapten to BSA (bovine serum albumin) was 1:2-20. Example 3

[0045] Etidomide was dissolved in 10% KOH methanol solution, stirred at room temperature, and the reaction was tracked by TLC. After complete reaction, the solvent was evaporated under reduced pressure, the residue was dissolved in water, and extracted twice with diethyl ether, the organic phase was removed, and the water phase was placed in an ice water bath and acidified with 1 mol / L hydrochloric acid to adjust the pH value to 6-7. The precipitated solid was filtered and dried to obtain the hydrolysis product as the etomidate hapten.

[0046] Take 10 mg etomidate hapten to dissolve in 0.5 mL DMF, add 8 mg NHS (N-hydroxysuccinimide), 10 mg EDC (carbonyldiimidazole), and stir until the reaction is complete. Then, take 50 mg KLH (keyhole limpet hemocyanin) to dissolve in 2 mL PBS (pH = 7.4), and drop the etomidate hapten derivative solution into the KLH solution while stirring. Continue the reaction for 12 h. The preparation method of the PBS solution is as follows: take 8.5 g NaCl, 0.3 g NaH2PO4, and 2.9 g Na2HPO4.12H2O, add water to make up to 1 L, and adjust the pH value to 7.4. Store the solution at 4 °C for standby use. Finally, perform low-temperature dialysis in PBS dialysis solution for 3 days. Store for immunization. The mass ratio of the etomidate hapten to KLH (keyhole limpet hemocyanin) is 1:2-20. Example 4

[0047] Preparation of etomidate immune antibodies, which specifically includes the following steps:

[0048] Preparation of polyclonal antibodies: immunize New Zealand white rabbits with etomidate complete antigen, and the immunization dose is 1 mg / time, with multiple point injections subcutaneously on the back.

[0049] First immunization: mix the etomidate complete antigen with an equal amount of Freund's complete adjuvant (FCA) and emulsify thoroughly;

[0050] Boost immunization: mix the etomidate complete antigen with an equal amount of Freund's incomplete adjuvant (FIA) and emulsify thoroughly, and perform the boost immunization 3 weeks after the first immunization. Continue the immunization for 4-5 times, with an interval of 2-3 weeks each time. Measure the titer of the immune antibodies by ELISA 10-15 days after the last immunization, and the titer should reach 10 5 At this time, collect blood and separate and collect serum. Extract IgG antibodies by saturated ammonium sulfate salting-out method, and store at -20 °C for standby use. Example 5

[0051] Preparation of etomidate monoclonal antibodies, which specifically includes the following steps:

[0052] After the immunization of the mice with the immunogen and the identification of the immunization, four 6-week-old Kunming mice were immunized with the immunogen, and after three booster immunizations, the titer was determined by blood sampling. When the serum titer no longer increased, the mice were immunized with a double dose of the immunogen without an adjuvant, and three days later, the mice were sacrificed by cervical dislocation, and the spleen was removed under sterile conditions. The spleen cells were mixed with the growing mouse myeloma cells at a ratio of 8:1 in a 50-mL centrifuge tube, 30 mL of serum-free IPMI1640 medium was added, and the mixture was centrifuged at 1100 r / min for 5 minutes. The supernatant was discarded, and the cell mass was gently shaken and placed in a 37-degree Celsius water bath. 1 mL of 50% (volume percentage) PEG-4000 was slowly added to the cells, and the dripping was completed within 1 minute, while the bottom precipitate was gently stirred. After 1 minute of standing, 1 mL of serum-free medium was slowly and evenly added along the wall of the tube for the first 30 seconds, and then 2 mL was added for the next 30 seconds. Then, 27 mL of the termination fusion process was quickly added, and the mixture was centrifuged at 1100 r / min for 5 minutes. The supernatant was discarded, and the cells were resuspended in HAT selective medium and added to a 96-well cell culture plate that had been coated with feeder cells. The plate was incubated at 37 degrees Celsius in a 5% CO2 environment. After 7 days, the HT medium was replaced, and when the number of hybrid cells in the wells reached more than 300, indirect ELISA (enzyme-linked immunosorbent assay) was used for screening. Wells with strong positive results, good inhibition effects, and vigorous cell growth were selected for limited dilution cloning. After more than 3 rounds of cloning and detection, the cells in the wells that were positive in all tests were hybridoma cells that secreted monoclonal antibodies. The hybridoma cells were expanded for the preparation of monoclonal antibodies. Then, the in vivo induction of ascites was used to produce etomidate monoclonal antibodies.

[0053] The steps of the in vivo induction of ascites are as follows: 0.5 mL of liquid paraffin oil was injected into the abdominal cavity of each mouse, and after 7 days, 3-5 x 10 6 / mouse was injected into the abdominal cavity of each mouse, and after 10 days, when the mouse's abdomen was significantly swollen, the ascites was collected. The ascites was purified by the n-octanoic acid-ammonium sulfate precipitation method to obtain the etomidate monoclonal antibodies. Example 6

[0054] As shown in FIGS. 1 and 2, an etomidate detection test strip, i.e., an etomidate colloidal gold chromatographic detection test strip, includes a backing plate 6, a sample pad 2, a reagent pad 3, a coated membrane, and a water-absorbing pad 5. The backing plate 6 is a PVC backing plate, the reagent pad 3 is a colloidal gold combined pad, the coated membrane is a nitrocellulose membrane 4, hereinafter referred to as an NC membrane, and the colloidal gold combined pad is an etomidate gold label combined pad of glass fibers on which etomidate colloidal gold-labeled antibodies are fixed. One end of the colloidal gold combined pad is connected to the NC membrane, and the other end is connected to the sample pad 2. One end of the sample pad 2 is connected to a liquid suction rod 1, and the other end of the coated membrane is connected to the water-absorbing pad 5. The detection line 8 (also referred to as the T line) and the quality control line 7 (also referred to as the C line) on the coated membrane are coated with etomidate complete antigen and goat anti-rabbit IgG antibodies, respectively.

[0055] In the detection, the sample is absorbed by the liquid-absorbing stick 1 and dropped on the sample position 9. The sample is uniformly supplied to the etomidate gold label binding pad through the sample pad 2 by capillary action, so that the label is rehydrated, reacts, and then moves along the liquid chromatography direction 10.

[0056] If the sample contains etomidate component, it will react with the etomidate colloidal gold labeled antibody. Then at least one red band is formed on the detection line on the NC membrane when passing through the detection area T line, which is judged as negative. Otherwise, when the T line is not colored, the result is positive. Due to the excess of the etomidate colloidal gold labeled antibody, the etomidate colloidal gold labeled antibody will be chromatographed to the C line, so that a color band is formed on the quality control line on the NC membrane. The color band is the standard for judging whether the chromatography process is normal and whether the reagent strip is deteriorated.

[0057] In practical application, the reagent strip of the present application is designed as a stick-shaped sample, the front end of which is connected with the sample pad for detection by a liquid-absorbing stick with strong water absorption capacity. The liquid-absorbing stick can absorb the sample, and the final sample is chromatographed through the NC membrane to complete the detection.

[0058] In practical application, the sample pad is a urine test special sample pad. Example 7

[0059] As shown in FIG. 1 and FIG. 2, an etomidate detection kit comprises the etomidate detection reagent strip shown in Example 6, further comprises a desiccant, and an aluminum foil bag for sealing and storing the etomidate detection reagent strip and the desiccant. Example 8

[0060] A preparation method of an etomidate detection reagent strip, the preparation method of the etomidate detection reagent strip comprising the following steps:

[0061] Step S100: preparation of etomidate gold labeled antibody and gold label binding pad;

[0062] Step S101: prepared by using the trisodium citrate reduction method. Measure 1000 ml of ultrapure water in a conical flask, heat to boiling, add 20 ml of 1% chloroauric acid (HAuCl4.3H2O, purchased from Sigma-Aldrich Company, item number: G4022) solution, heat to vigorous boiling, add 20 ml of 2% trisodium citrate solution, continue to heat for 15 minutes after the color turns red, cool to room temperature, and then dilute to 1 L with ultrapure water, and store in a 4°C refrigerator. The OD value is detected by using an ultraviolet spectrophotometer, the maximum absorption peak is at 540±10 nm, and the size of the colloidal gold is 40 nm.

[0063] Step S102: Take the prepared colloidal gold 100 mL in a beaker, stir, add 1 ml of 0.1M K2CO3 and continue stirring for 30 minutes, then add 1.0 mg of etomidate antibody (0.8 mg-1.4 mg), stir for 60 minutes, add 2.0 ml of 10% BSA, continue stirring for 30 minutes, centrifuge the labeled gold at 6000 rpm for 15 minutes at 4-8°C, collect the precipitate, continue to centrifuge the supernatant at 8400 rpm for 30 minutes at 4-8°C, continue to centrifuge the supernatant for the third time at 9600 rpm for 30 minutes at 4-8°C, collect the precipitate. Mix the three collected precipitates, measure the absorbance value OD at 540 nm, and store in a refrigerator at 2-8°C for use.

[0064] Step S103: Dilute the etomidate antibody gold label solution with a gold label diluent (1L solution contains Tris 8.48g, BSA 10g, sucrose 100g, trehalose 50g, Triton X-100 4ml, water 1L, pH=8.2±0.1), evenly coat on glass fiber, and prepare etomidate gold label binding pad.

[0065] In actual application, the step S103 can also be realized by the following method: adsorb the etomidate colloidal gold labeled antibody diluted by 1:100-500 (diluted with 1% BSA, 0.5% Casein Na, 1% Triton x-100) in refined glass fiber cotton, dry at 37°C, and prepare etomidate gold label binding pad.

[0066] Step S200: Preparation of antigen-coated;

[0067] Spray the selected concentration (0.1-1.0 mg / mL) of coated antigen (dilute the complete antigen to a concentration of 0.1-1.0 mg / mL with 0.01M PBS buffer pH 7.2-7.6 or 1% BSA or 0.5% Casein Na or 1% Triton x-100) and goat anti-rabbit IgG (dilute the immune antibody to a concentration of 0.2-1.0 mg / mL with 0.01M PBS buffer pH 7.2-7.6 or 1% BSA or 0.5% Casein Na or 1% Triton x-100) on the coated membrane, the spraying amount is 1 μl / cm, and they are used as the detection line T and the control line C, respectively, and dried in a 37°C oven for 10 min.

[0068] Step S300: Assembly of reagent strip;

[0069] The test strip of PVC material is composed of a backing plate 6, on which a sample pad 2, a colloidal gold combined pad, a coating film and a water absorption pad 5 are sequentially adhered. The adhered plate is cut into a test strip with a width of 3 mm. The coating film is an NC film, and the sample pad is a urine test special sample pad with a size of 21 cm x 30 cm.

[0070] In practical applications, the prepared etomidate detection reagent strip needs to be subjected to a sensitivity test through the following steps:

[0071] The etomidate is prepared into various concentrations shown in the following table by using a diluent (the diluent is a 0.01M PBS buffer solution with a pH of 7.2-7.6 or 1% BSA or 0.5% Casein Na or 1% Triton x-100), and the results are detected and observed. The results are shown in the following table:

[0072] Etomidate detection results

[0073]

[0074] Note: "+" represents positive; "-" represents negative.

[0075] The experimental results show that the test paper has a minimum detection amount of 100 ng / mL for etomidate.

[0076] Specificity

[0077] A 100 μg / ml detection sample is prepared, and the detection results are as follows:

[0078]

[0079] Cross reaction

[0080] The reagent can also detect the homolog of etomidate, i.e. metomidate, with a sensitivity of 5 μg / ml.

[0081] Example 9

[0082] An etomidate detection method, which uses the etomidate reagent strip as described in Examples 6-8 for detection, specifically includes the following steps:

[0083] Step S01: horizontally place the etomidate reagent strip;

[0084] Step S02: drop 3-4 drops of the sample to be detected;

[0085] Step S03: observe and determine the detection results for 5 minutes;

[0086] Step S04: determine the results.

[0087] If only a red line of C line appears on the coated film, it indicates that the detection result is positive, if two red lines of C line and T line both appear on the coated film, it indicates that the detection result is negative, and if the red line of C line does not appear on the coated film, it indicates that the test strip has failed.

[0088] The advantages of the embodiment of the present application are: by the above-mentioned scheme, the characteristic structure of etomidate is retained to the maximum extent, and the etomidate monoclonal antibody prepared by further utilizing the etomidate hapten has good affinity, sensitivity and specificity, and the titer of the immune serum obtained by immunization is as high as 10 5 The above detection is rapid and accurate.

[0089] The above is only a specific embodiment of the present application, but the protection scope of the present application is not limited thereto, any person skilled in the art can easily think of changes or replacements within the technical range disclosed by the present application, which should be covered within the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. A method for preparing a complete antigen of etomidate, characterized by, The preparation method of the etomidate complete antigen comprises the following steps: The etomidate hapten derivative solution is prepared by the etomidate hapten; The carrier protein solution is prepared by dissolving the carrier protein; The etomidate complete antigen is prepared by mixing the etomidate hapten derivative solution and the carrier protein solution for coupling reaction.

2. The method for preparing etomidate complete antigen according to claim 1, characterized in that, The etomidate hapten is prepared by reacting etomidate with a methanol solution under alkaline conditions to obtain an intermediate, and then hydrolyzing and extracting the intermediate.

3. The method for preparing etomidate complete antigen according to claim 1, characterized in that, The etomidate hapten derivative solution is prepared by dissolving the etomidate hapten in an organic solvent, adding N-hydroxysuccinimide and carbodiimide, and stirring for reaction.

4. The method for preparing etomidate complete antigen according to claim 1, characterized in that, The carrier protein solution is prepared by dissolving the carrier protein in a buffer solution, wherein the carrier protein is selected from bovine serum albumin or hemocyanin, and the buffer solution is a PBS buffer solution.

5. The method for preparing etomidate complete antigen according to claim 1, characterized in that, The mass ratio of the etomidate hapten derivative to the carrier protein is 1:2-20.

6. The method for preparing etomidate complete antigen according to claim 1, characterized in that, After the coupling reaction, low-temperature dialysis is performed, and the low-temperature dialysis time is 1-3 days.

7. A complete antigen of etomidate, characterized in that, The etomidate complete antigen is prepared by the preparation method of the etomidate complete antigen according to any one of claims 1-6.

8. Any one of the following applications of the etomidate complete antigen according to claim 7: for detecting etomidate and its homologues; for preparing an immunochromatographic reagent strip of etomidate; for preparing an etomidate detection test paper or kit.

9. A etomidate immunochromatographic reagent strip comprising a substrate, a sample application part, a detection part, and a liquid absorption part, characterized in that, The sample adding component is adsorbed with colloidal gold labeled etomidate clone antibody prepared by the etomidate complete antigen, the detection component is fixed on a substrate, a quality control band and a detection band are arranged in the middle of the detection component, and liquid suction components and sample adding components are fixed at both ends of the detection component in a partially overlapping manner, wherein the detection band is coated with the etomidate complete antigen according to claim 7, and the quality control band is coated with goat anti-rabbit IgG.

10. The etomidate immunochromatographic reagent strip according to claim 9, characterized by, The substrate is a PVC lining board, and the detection component is a nitrocellulose membrane.

11. A test kit for etomidate and its homologues, characterized in that, The detection kit comprises the etomidate immunochromatographic reagent strip according to claim 9 or 10, and an etomidate detection analysis solution matched with the etomidate immunochromatographic reagent strip according to claim 9 or 10.

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