Kit and method for detecting content of sertraline

Abstract

The invention provides a kit and a method for detecting the content of sertraline, and belongs to the technical field of in-vitro detection. The kit comprises the following components: (1) a reagent R1, wherein the reagent R1 comprises a sertraline antigen combined with a luminescent substance; (2) a reagent M1, wherein the reagent M1 comprises a sertraline antibody coupled with a fluorescent marker; wherein the structure of the sertraline antigen is shown as a formula C, and the sertraline antibody is an antibody generated in response to the sertraline antigen. The kit disclosed by the invention can be used for rapidly and quantitatively detecting the content of the sertraline medicine in samples such as blood, urine and saliva, is wide in detection concentration range, high in precision, good in repeatability and simple and convenient to operate, and better meets the requirements of clinical popularization. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the technical field of in vitro detection, and particularly relates to a kit and a method for detecting the content of sertraline. Background Art

[0002] Sertraline is a first-line drug for the treatment of depression, and it is mainly metabolized into inactive N-desmethylsertraline in the liver. The effective blood drug concentration is 15 - 150 ng / mL, and when the blood drug concentration exceeds 300 ng / mL, toxic reactions are likely to occur. Based on this, it is very necessary to monitor the blood drug concentration of sertraline in patients with depression to supervise clinical medication, formulate a reasonable dosing plan, determine the optimal treatment dose, ensure individualized medication, improve the curative effect and reduce adverse reactions.

[0003] Currently, the commonly used methods for detecting sertraline are gas chromatography-mass spectrometry (GCMS), high performance liquid chromatography (HPLC), homogeneous enzyme immunoassay (EMIT), etc. However, these methods are complex in operation and have certain limitations in large-scale clinical promotion and application. At present, there are few sertraline detection reagents with good stability, high sensitivity and strong specificity on the market.

[0004] The patent with the publication number CN110456087B discloses a sertraline detection reagent and its preparation and use method, and this patent discloses a sertraline maleimide derivative

[0005] and uses this sertraline maleimide derivative to prepare a sertraline immunogen with high immunogenicity, and then immunizes experimental animals. Blood is taken from the immunized experimental animals, and a sertraline-specific antibody is obtained by separation and purification; meanwhile, a sertraline enzyme-labeled conjugate is prepared using this sertraline maleimide derivative. The sertraline detection reagent containing the above-mentioned sertraline-specific antibody and sertraline enzyme-labeled conjugate can achieve high-throughput and rapid detection of sertraline on an automatic biochemical analyzer. However, the detection range of the sertraline detection kit reported in this patent is narrow and it is difficult to meet the clinical needs. Summary of the Invention

[0006] Currently, there are very few products that can quantitatively detect sertraline by immunological methods. The purpose of the present invention is to provide a kit for detecting sertraline, and a method for detecting the content of sertraline by chemiluminescence immunoassay.

[0007] The present invention provides a kit for detecting the content of sertraline, which comprises the following components:

[0008] (1) Reagent R1, and the reagent R1 includes a sertraline antigen combined with a luminescent substance;

[0009] (2) Reagent M1, where the reagent M1 includes a sertraline antibody conjugated with a fluorescent label;

[0010] Among them, the structure of the sertraline antigen is as shown in formula C:

[0011]

[0012] Among them, L is none or a linker, R2 is a carrier protein, and n is an integer from 1 to 10;

[0013] The sertraline antibody is an antibody produced in response to the sertraline antigen.

[0014] Furthermore, the structure of the sertraline antigen is as follows:

[0015]

[0016] Among them, L is CONH or NHCO, and R2 is a carrier protein.

[0017] Furthermore, the carrier protein is keyhole limpet hemocyanin, ovalbumin or bovine serum albumin.

[0018] Furthermore, the sertraline antibody is a monoclonal antibody or a polyclonal antibody.

[0019] Furthermore, the preparation method of the reagent R1 includes the following steps: mixing the sertraline antigen solution and the luminescent substance solution, reacting for 1 - 3 h, adding a termination solution to terminate the reaction, filtering, and diluting to obtain the reagent R1;

[0020] The preparation method of the reagent M1 includes the following steps: activating the sertraline antibody with an activator, mixing the activated sertraline antibody with a fluorescent label, reacting for 30 - 60 min to obtain a fluorescent label - labeled sertraline antibody, adding a blocking solution and reacting for 30 - 60 min, washing, and suspending to obtain the reagent M1.

[0021] Furthermore, in the preparation method of the reagent R1, the concentration of the sertraline antigen solution is 0.1 - 5 mg / mL; the concentration of the luminescent substance solution is 1 - 10 mmol / L; the volume ratio of the sertraline antigen solution to the luminescent substance solution is (20 - 30):1; the luminescent substance is acridinium ester; the dilution factor is 1000 - 4000 times;

[0022] In the preparation method of the reagent M1, the molar ratio of the sertraline antibody to the activator is 1:(10 - 30); the activator is biotin ester; the mass ratio of the activated sertraline antibody to the fluorescent label is 0.1:(50 - 150); the fluorescent label is magnetic microparticles; in the reagent M1, the concentration of the fluorescent label is 0.1 - 1.0 mg / mL.

[0023] Furthermore, in the preparation method of reagent R1, the concentration of the sertraline antigen solution is 1 mg / mL; the concentration of the luminescent substance solution is 5 mmol / L; the volume ratio of the sertraline antigen solution to the luminescent substance solution is 25:1; the dilution factor is 2000-fold;

[0024] In the preparation method of reagent M1, the molar ratio of the sertraline antibody to the activator is 1:20; the mass ratio of the activated sertraline antibody to the fluorescent label is 0.1:100; in reagent M1, the concentration of the fluorescent label is 0.4 mg / mL.

[0025] Furthermore, in the preparation method of reagent R1, the reaction condition is to react at room temperature in the dark for 2 h; the termination solution is lysine solution or Tris buffer;

[0026] In the preparation method of reagent M1, the blocking solution is a 3 g / L biotin solution, the washing is carried out using Tris-HCl buffer, and the suspension is carried out using Tris-HCl buffer.

[0027] Furthermore, the kit further comprises a sertraline calibrator.

[0028] The present invention also provides a method for detecting the content of sertraline, which is to detect the content of sertraline in a sample to be detected by using the above kit.

[0029] Furthermore, the method comprises the following steps: performing chemiluminescence detection using the sample to be detected, reagent R1 and reagent M1 to obtain a detection signal value, and converting the detection signal value into the content of sertraline in the sample to be detected according to a standard curve.

[0030] Furthermore, the standard curve is obtained by the following method: performing chemiluminescence detection using the sertraline calibrator, reagent R1 and reagent M1 to obtain a detection signal value, and plotting a standard curve.

[0031] Furthermore, the sample to be detected is blood, urine or saliva.

[0032] Furthermore, the volume ratio of the sertraline calibrator, reagent R1 and reagent M1 is 1:10:5, and the volume ratio of the sample to be detected, reagent R1 and reagent M1 is 1:10:5.

[0033] When detecting the content of sertraline by using the method of the present invention, an external sertraline calibrator can be used to plot a standard curve, or an existing standard curve can be directly adopted.

[0034] As used herein, the term "antibody" refers not only to intact antibody molecules, but also to antibody fragments or derivatives that retain the specific binding ability of intact antibodies. The antibodies referred to in the present invention can be monoclonal antibodies or polyclonal antibodies.

[0035] The present invention designs a new sertraline derivative. After conjugating the sertraline derivative with a carrier protein, a sertraline antigen is obtained. Using this sertraline antigen for animal immunization yields monoclonal antibodies with high specificity and sensitivity.

[0036] Using the kit of the present invention, which includes a sertraline calibrator, a sertraline antigen conjugated with a luminescent substance, and a sertraline antibody conjugated with a fluorescent label, can rapidly and quantitatively detect the content of sertraline drug in samples such as blood, urine, and saliva. The detection has good repeatability and is easy to operate. Moreover, compared with the detection method disclosed in CN110456087B, the detection method using the kit of the present invention has a wider detection concentration range (up to 200 ng / mL), higher precision, and better meets the requirements of clinical promotion.

[0037] Obviously, based on the above content of the present invention, according to the common general knowledge and conventional means in the art, without departing from the above basic technical idea of the present invention, various other forms of modifications, substitutions, or changes can be made.

[0038] The above content of the present invention will be further described in detail below through specific embodiments in the form of examples. However, this should not be construed as limiting the scope of the above subject matter of the present invention to the following examples. All technologies implemented based on the above content of the present invention fall within the scope of the present invention.

[0039] Description of the Drawings

[0040] Figure 1 : Detection result of the liquid chromatography-mass spectrometry (LC-MS) of Compound 4.

[0041] Figure 2 : 1 1H NMR detection result of Compound 4.

[0042] Figure 3 : HPLC detection result of Compound 4.

[0043] Figure 4 : Standard curve for sertraline content detection. Detailed Description of the Invention

[0044] The raw materials and equipment used in the present invention are all known products, obtained by purchasing commercially available products.

[0045] In the operations of the examples of the present invention, room temperature refers to 10 - 30 °C, and the overnight time refers to 8 - 16 h.

[0046] Example 1, Sertraline Rapid Quantitative Detection Kit

[0047] The sertraline rapid quantitative detection kit consists of the following components:

[0048] (I) Calibrator; the calibrator is commercially available sertraline.

[0049] (II) M1 reagent;

[0050] (III) R1 reagent.

[0051] The volume ratio of the calibrator, M1 reagent, and R1 reagent is 1∶5∶10.

[0052] The preparation method of the M1 reagent is as follows:

[0053] (a) Biotinylated antibody: Mix the sertraline antibody and biotin ester at a molar ratio of 1∶20, react for 40 min, and add 1 mol / L lysine to terminate the reaction to obtain a biotinylated antibody solution;

[0054] (b) Dialysis: Dialyze the biotinylated antibody solution obtained in step (1) for 24 - 48 h, recover the biotinylated antibody and add the same volume of glycerol for standby;

[0055] (c) Magnetization: Wash the magnetic particles with Tris-HCl buffer solution 2 - 5 times; mix the biotinylated antibody and magnetic particles at a mass ratio of 0.1∶100 and react at room temperature for 40 min;

[0056] (d) Blocking: Add 10 μL of blocking solution according to 1 mg of magnetic particles, add the blocking solution and react at room temperature for 40 min; the blocking solution is a 3 g / L vitamin H solution;

[0057] (e) Washing: Wash with Tris-HCl buffer solution 2 - 5 times;

[0058] (f) Suspension: Suspend with Tris-HCl buffer solution, and the concentration of magnetic particles in the suspension is 0.4 mg / mL.

[0059] The preparation method of the R1 reagent is as follows:

[0060] (i) Mix the sertraline antigen solution with a concentration of 1 mg / mL and the acridinium ester solution with a concentration of 5 mmol / L at a volume ratio of 25∶1, and react in the dark at room temperature for 2 h;

[0061] (ii) Add 1 mol / L lysine to terminate the reaction for 30 min;

[0062] (iii) Filter and purify the reaction mixture with a filter column for standby;

[0063] (iv) Dilute it with PBS buffer at a volume ratio of 1:2000 to obtain Reagent R1.

[0064] The following are the preparation methods of sertraline antigen and sertraline antibody:

[0065] I. Preparation of sertraline derivative

[0066]

[0067] (1) Compound 1 and Compound 2 undergo a substitution reaction to obtain Compound 3:

[0068]

[0069] Weigh Compound 1 (1.0 g, 3.26 mmol, 1.0 eq) and dissolve it in dimethylformamide (15 mL). Add Compound 2 (709.4 mg, 4.0 mmol, 1.2 eq), K2CO3 (1.35 g, 9.76 mmol, 3.0 eq) and KI (200 mg, 1.2 mmol). Heat to 100 °C and stir overnight. Check by TLC that Compound 1 has no residue. Add water (30 ml) for dilution, extract with ethyl acetate (30 mL × 3). Combine the organic phases, wash once with saturated brine, dry with anhydrous sodium sulfate, concentrate, and obtain Compound 3 (520 mg, 40%) by column chromatography, which is a transparent oil.

[0070] (2) Compound 3 undergoes a demethylation reaction to obtain Compound 4:

[0071]

[0072] Weigh Compound 3 (500 mg, 1.23 mmol) and dissolve it in a mixed solution of THF (10 mL) and MeOH (10 mL). Add LiOH (1 M, 5 mL) to provide an alkaline environment and stir at room temperature overnight. Check by TLC that Compound 3 has no residue to obtain the crude product of Compound 4. Adjust the pH of the crude product of Compound 4 to 5 with 2M HCl, extract 3 times with ethyl acetate at 0 °C, wash the separated organic phase with saturated brine, and dry with anhydrous sodium sulfate to obtain Compound 4 (220 mg, 46%), which is a white solid.

[0073] The structure characterization data of Compound 4 are shown in Figures 1-3 and below:

[0074] LCMS: RT = 1.07 min, [MS + 1] = 392.0, [MS - 1] = 390.0.

[0075] 1 1H NMR: 11H NMR (400 MHz, Methanol-d4) δ 7.79 (d, J = 7.8 Hz, 1H), 7.45 - 7.31 (m, 3H), 7.23 (d, J = 2.1 Hz, 1H), 7.06 (d, J = 7.6 Hz, 1H), 6.98 (dd, J = 8.4, 2.2 Hz, 1H), 4.76 - 4.74 (m, 1H), 4.26 (br s, 1H), 3.20 - 3.09 (m, 2H), 2.63 (s, 3H), 2.47 - 2.46 (m, 2H), 2.18 (br s, 2H), 2.03 - 1.27 (m, 4H).

[0076] II. Preparation of sertraline antigen

[0077] The sertraline antigen is a conjugate formed by coupling a sertraline derivative and a carrier protein. The specific preparation method is as follows:

[0078] (i) Dissolve the sertraline derivative in a dimethylformamide solution to obtain a sertraline derivative solution, where the concentration of the sertraline derivative is 5×10 -5 mg / μL.

[0079] (ii) Weigh 5 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) and 1 mg of N-hydroxysuccinimide (NHS) and add them to 1 L of the sertraline derivative solution, and react in the dark for 1 - 4 hours to obtain a reaction solution.

[0080] (iii) Weigh 10 mg of ovalbumin and dissolve it in 5 mL of 0.01 M PBS (pH 7.4) solution to obtain a colorless, clear and transparent ovalbumin buffer solution; mix the reaction solution in step (ii) with the ovalbumin buffer solution to obtain a slightly milky turbid solution, and react in the dark at room temperature overnight to obtain a slightly milky turbid liquid.

[0081] (iv) Centrifuge the turbid liquid (6000 r / min, 5 min), and extract the supernatant;

[0082] (v) Dialyze the supernatant with PBS buffer solution for 2 d, the pH of the PBS buffer solution is 7.4, and the dialysis temperature is 2 - 8 °C;

[0083] (vi) Centrifuge the retained solution after dialysis (6000 r / min, 5 min), and extract the supernatant to obtain the sertraline-ovalbumin complex, that is, the sertraline antigen.

[0084] The structure of the obtained sertraline antigen is: R 2b is ovalbumin.

[0085] III. Preparation of sertraline antibody

[0086] The sertraline antibody is obtained by immunizing animals with sertraline antigen. The specific immunization method is as follows:

[0087] a. After immunizing animals with sertraline antigen by the conventional Freund's adjuvant method for multiple times, serum containing specific antibodies is obtained. After the titer is qualified, spleen cells are taken for fusion.

[0088] b. Screen positive fused cells and perform subcloning to obtain monoclonal cell lines.

[0089] c. Produce ascites or produce by cell culture, and isolate and purify to obtain monoclonal antibodies.

[0090] Test the titer and IC 50 value of the above sertraline antibody.

[0091] 1. Titer test sample

[0092] The sertraline antibody prepared in Example 5.

[0093] 2. Titer test method

[0094] Use the ELISA method to detect the titer and half-inhibitory concentration (IC 50 ) of the sertraline antibody. The specific operation is as follows:

[0095] (2.1) Instruments:

[0096] Electric thermostatic forced air drying oven, electronic balance, fully automatic microplate reader, fully automatic plate washer.

[0097] (2.2) Reagents

[0098] The sertraline antibody prepared in Example 5;

[0099] HRP-labeled goat anti-mouse enzyme-labeled secondary antibody;

[0100] TMB chromogenic solution (when in use, the TMB chromogenic solution needs to be restored to room temperature. Note: Place it in an environment of 18-28 °C for 30-45 min);

[0101] 0.1M PH = 9.6 sodium bicarbonate buffer solution;

[0102] 0.01M PBS;

[0103] Stop solution (2M H2SO4 solution or 1M HCl solution);

[0104] Standard solution.

[0105] (2.3) Method

[0106] (2.3.1) Indirect ELISA (detecting antigen and antibody titers)

[0107] a) Dilute the antigen to be tested (or qualified antigen) with 0.1M sodium bicarbonate buffer at pH 9.6 to 1 μg / mL, add it to a 96-well ELISA plate, 100 μL per well, react at 37 °C for 3 h or stand overnight at 4 °C.

[0108] b) Discard the liquid in the wells of the plate, add 250 μL of ELISA washing solution, stand for 30 s, discard the liquid in the plate, and repeat 3 times. (Wash away the uncoated free antigen).

[0109] c) Dilute the qualified antibody (or antibody to be tested) with 0.01M PBS to 1 mg / mL, and then dilute it again with 0.01M PBS to 1k, 2k, 4k, 8k, 16k, 32k, 64k in labeled centrifuge tubes for standby.

[0110] d) Add the qualified antibody (or antibody to be tested) diluted in the above steps (negative control is 0.01M PBS), 100 μL per well, make three replicates for each concentration gradient, and react at 37 °C for 45 min.

[0111] e) Discard the liquid in the wells of the plate, add 250 μL of ELISA washing solution, stand for 30 s, discard the liquid in the plate, and repeat 3 times.

[0112] f) Dilute the HRP-labeled goat anti-mouse enzyme-labeled secondary antibody according to the instructions (dilute with 0.01M PBS), add the diluted HRP-labeled goat anti-mouse enzyme-labeled secondary antibody, 100 μL per well, and react at 37 °C for 45 min.

[0113] g) Discard the liquid in the wells of the plate, add 250 μL of ELISA washing solution, stand for 30 s, discard the liquid in the plate, and repeat 3 times.

[0114] h) Add the chromogenic solution (protected from light) (TMB chromogenic solution needs to be restored to room temperature), 100 μL per well, and react at room temperature (18 - 28 °C environment) in the dark for 15 min.

[0115] i) Add the stop solution (1M HCl solution or 2M H2SO4 solution), 100 μL per well, OD 450 Read the OD value.

[0116] Note: 1M HCl solution is prepared from 6M HCl solution, prepared and used immediately. Taking 12 mL of the preparation amount as an example, add 2 mL of 6M HCl solution to 10 mL of purified water and vortex to mix evenly.

[0117] (2.3.2) Competitive ELISA (detecting antigen-antibody competition)

[0118] a) Dilute the antigen to be tested (or qualified antigen) to 1 μg / mL with 0.1 M sodium bicarbonate buffer at pH 9.6, add it to a 96-well ELISA plate, 100 μL per well, react at 37 °C for 3 h or let it stand overnight at 4 °C.

[0119] b) Discard the liquid in the wells of the plate, add 250 μL of ELISA washing solution, let it stand for 30 s, discard the liquid in the plate, and repeat 3 times. (Wash away the uncoated free antigen)

[0120] c) Dilute the qualified antibody (or antibody to be tested) to 1 mg / mL with 0.01 M PBS, and then dilute it again to 1k, 2k, 4k, 8k, 16k, 32k, 64k with 0.01 M PBS in labeled centrifuge tubes for standby.

[0121] d) Add 50 μL of the diluted standard solution (negative control is 0.01 M PBS) to each well, and then add 50 μL of the diluted qualified antibody (or antibody to be tested) to each well (negative control is 0.01 M PBS). Make three replicates for each concentration gradient and react at 37 °C for 45 min.

[0122] e) Discard the liquid in the wells of the plate, add 250 μL of ELISA washing solution, let it stand for 30 s, discard the liquid in the plate, and repeat 3 times.

[0123] f) Dilute the HRP-labeled goat anti-mouse enzyme-linked secondary antibody according to the instructions (dilute with 0.01 M PBS), add the diluted HRP-labeled goat anti-mouse enzyme-linked secondary antibody, 100 μL per well, and react at 37 °C for 45 min.

[0124] g) Discard the liquid in the wells of the plate, add 250 μL of ELISA washing solution, let it stand for 30 s, discard the liquid in the plate, and repeat 3 times.

[0125] h) Add the chromogenic solution (store in the dark) (TMB chromogenic solution needs to be restored to room temperature), 100 μL per well, and react in the dark at room temperature (18 - 28 °C environment) for 15 min.

[0126] i) Add the stop solution (1 M HCl solution or 2 M H2SO4 solution), 100 μL per well, OD 450 Read the OD value.

[0127] Note: The 1 M HCl solution is prepared from 6 M HCl solution and should be prepared and used immediately. Taking 12 mL of the preparation amount as an example, add 2 mL of 6 M HCl solution to 10 mL of purified water and vortex to mix evenly.

[0128] Finally, calculate the IC according to the ELISA standard curve 50 .

[0129] 3. Test results

[0130] Table 1. Results of the inhibition assay for sertraline antibodies

[0131]

[0132] It can be seen that the sertraline antibody of the present invention can specifically bind to sertraline.

[0133] Table 2. Inhibition rates of sertraline antibodies at different concentrations (antibody dilution factor is 2K)

[0134]

[0135] As shown in Table 2, the IC 50 value of the sertraline antibody of the present invention is 1.94 ng / mL.

[0136] Example 2. Rapid quantitative detection kit for sertraline

[0137] The rapid quantitative detection kit for sertraline consists of the following components:

[0138] (I) Calibrator; the calibrator is commercially available sertraline.

[0139] (II) M1 reagent;

[0140] (III) R1 reagent.

[0141] The volume ratio of the calibrator, M1 reagent, and R1 reagent is 1:5:10.

[0142] Compared with the rapid quantitative detection kit for sertraline in Example 1, the difference in the detection kit of this example is only that: 1) ovalbumin in the sertraline antigen is replaced with bovine serum albumin, and the structure of the sertraline antigen is: R 2a is bovine serum albumin; 2) the sertraline antibody is obtained by immunizing animals with the aforementioned sertraline antigen.

[0143] Example 3. Method for detecting sertraline content

[0144] The chemiluminescent immunoassay for sertraline is carried out using the rapid quantitative detection kit for sertraline in Example 1. The specific steps are as follows:

[0145] (1) Mix the calibrator, M1 reagent, and R1 reagent according to the volume ratio of 1:5:10, perform luminescence detection to obtain the detection signal value, and draw a standard curve;

[0146] (2) Mix the test sample, M1 reagent, and R1 reagent according to the volume ratio of 1:5:10, perform luminescence detection to obtain the detection signal value;

[0147] (3) Convert the detected signal value according to the standard curve to obtain the content of sertraline in the sample to be tested.

[0148] The test results of calibration standards with different concentrations are shown in Table 4, and the standard curve plotted is as Figure 4 shown.

[0149] Table 4. Test Results of Calibration Standards with Different Concentrations

[0150]

[0151] The test results of the content of sertraline in the sample to be tested are shown in Table 5.

[0152] Table 5. Test Results of the Content of Sertraline in the Sample to be Tested

[0153] Sample 1 (ng / mL) Sample 2 (ng / mL) Data 1 10.11 149.85 Data 2 10.61 152.25 Data 3 10.25 151.12 Data 4 10.40 150.53 Data 5 10.72 150.64 Data 6 10.67 150.17 Data 7 10.16 151.51 Data 8 10.06 151.79 Data 9 10.30 151.48 Data 10 10.46 150.86 Average 10.38 151.02 Standard Deviation (SD) 0.24 0.75 Coefficient of Variation (CV) 2.28％ 0.50％

[0154] The above results show that the kit of the present invention has a wide detection concentration range up to 200 ng / mL when detecting the content of sertraline. The coefficient of variation (CV) of detecting the content of sertraline by using the kit of the present invention is low, indicating that the precision of detecting the content of sertraline by using the kit of the present invention is high, and it can accurately detect the content of sertraline in the sample to be tested.

[0155] In summary, the present invention provides a kit for detecting sertraline, and a method for detecting the content of sertraline by chemiluminescent immunoassay. The kit of the present invention can quickly and quantitatively detect the content of sertraline drug in samples such as blood, urine, saliva, etc., has a wide detection concentration range, high precision, good repeatability, simple operation, and better meets the requirements of clinical promotion.

Claims

1. A kit for detecting the content of sertraline, characterized in that, It comprises the following components: (1) Reagent R1, wherein the reagent R1 comprises sertraline antigen conjugated with a luminescent substance; (2) Reagent M1, wherein the reagent M1 comprises sertraline antibody conjugated with a fluorescent label; Among them, the structure of the sertraline antigen is as shown in Formula C: Among them, L is none or a linker, R2 is a carrier protein, and n is an integer from 1 to 10; The sertraline antibody is an antibody produced in response to the sertraline antigen.

2. The kit according to claim 1, wherein The structure of the sertraline antigen is as follows: Among them, L is CONH or NHCO, and R2 is a carrier protein.

3. The kit according to claim 2, characterized in that, The carrier protein is keyhole limpet hemocyanin, ovalbumin or bovine serum albumin; And / or, the sertraline antibody is a monoclonal antibody or a polyclonal antibody.

4. The kit according to any one of claims 1-3, characterized in that The preparation method of the reagent R1 comprises the following steps: mixing the sertraline antigen solution with the luminescent substance solution, reacting for 1 - 3 h, adding a termination solution to terminate the reaction, filtering, and diluting to obtain the reagent R1; The preparation method of the reagent M1 comprises the following steps: activating the sertraline antibody with an activator, mixing the activated sertraline antibody with the fluorescent label, reacting for 30 - 60 min to obtain the sertraline antibody labeled with the fluorescent label, adding a blocking solution and reacting for 30 - 60 min, washing, and suspending to obtain the reagent M1.

5. The preparation method of the kit according to claim 4, characterized in that, In the preparation method of the reagent R1, the concentration of the sertraline antigen solution is 0.1 - 5 mg / mL; the concentration of the luminescent substance solution is 1 - 10 mmol / L; the volume ratio of the sertraline antigen solution to the luminescent substance solution is (20 - 30):1; the luminescent substance is acridinium ester; the dilution factor is 1000 - 4000 times; In the preparation method of the reagent M1, the molar ratio of the sertraline antibody to the activator is 1:(10 - 30); the activator is biotin ester; the mass ratio of the activated sertraline antibody to the fluorescent label is 0.1:(50 - 150); the fluorescent label is magnetic microparticles; in the reagent M1, the concentration of the fluorescent label is 0.1 - 1.0 mg / mL.

6. The preparation method of the kit according to claim 5, characterized in that, In the preparation method of the reagent R1, the concentration of the sertraline antigen solution is 1 mg / mL; the concentration of the luminescent substance solution is 5 mmol / L; the volume ratio of the sertraline antigen solution to the luminescent substance solution is 25:1; the dilution factor is 2000 times; In the preparation method of the reagent M1, the molar ratio of the sertraline antibody to the activator is 1:20; the mass ratio of the activated sertraline antibody to the fluorescent label is 0.1∶100; in the reagent M1, the concentration of the fluorescent label is 0.4 mg / mL.

7. The kit according to any one of claims 1-6, characterized in that, The kit further comprises a sertraline calibrator.

8. A method for detecting the content of sertraline, characterized in that, The method is to detect the content of sertraline in a sample to be detected by using the kit according to any one of claims 1 - 7.

9. The method according to claim 8, wherein The method comprises the following steps: performing chemiluminescence detection using the sample to be detected, reagent R1 and reagent M1 to obtain a detection signal value, and converting the detection signal value into the content of sertraline in the sample to be detected according to a standard curve.

10. The method according to claim 9, wherein, The standard curve is obtained by the following method: Using sertraline calibrator, reagent R1 and reagent M1 to perform chemiluminescence detection to obtain detection signal values, and plotting the standard curve; And / or, the sample to be tested is blood, urine or saliva.

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