Escitalopram detection kit and detection method

Abstract

The invention belongs to the technical field of in-vitro detection, and particularly relates to an escitalopram detection kit and a detection method. According to the present invention, the kit comprising the escitalopram antigen and the escitalopram antibody is constructed, and the immunological detection of escitalopram can be achieved by using the kit. The detection kit and the detection method disclosed by the invention are high in accuracy and good in repeatability, and have a very good application prospect.

Description

Technical Field

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

[0002] Escitalopram oxalate is a selective serotonin reuptake inhibitor (SSRIs). Due to its significant effect and low side effects, it has become a first-line drug for current antidepressant treatment. Clinical applications have found that there are very large individual differences in the efficacy of escitalopram oxalate drugs. This difference mainly stems from the difference in the activity of drug-metabolizing enzymes. By monitoring the blood drug concentration, it helps to judge whether the drug concentration in the patient's body is within the therapeutic window, ensuring the efficacy while minimizing the occurrence of adverse reactions as much as possible. In addition, by monitoring the blood drug concentration, it can also judge whether the current blood drug concentration of the patient is consistent with the dosage, so as to understand whether the patient takes the medicine according to the doctor's advice.

[0003] According to the Consensus Guidelines for Therapeutic Drug Monitoring in Neuropsychopharmacology: Update 2017, there is a clear effective therapeutic concentration range of 15 - 80 ng / mL for the detection of escitalopram, and the warning concentration is 160 ng / mL.

[0004] For the detection of escitalopram blood drug concentration, the existing mainstream methods are liquid chromatography-tandem mass spectrometry (LC-MS / MS) method, high performance liquid chromatography (HPLC), homogeneous enzyme immunoassay (EMIT), etc. Among them, the LC-MS / MS method has complex sample pretreatment, high requirements for testers, and expensive instrument equipment, which is not conducive to clinical promotion.

[0005] Immunological detection methods mainly include the detection of antigen-antibody reactions, such as ELISA, immunofluorescence, immunohistochemistry, etc., and the detection of lymphocyte functions, such as lymphocyte proliferation test, cytokine determination, etc. These methods are widely used in the fields of medical pathological diagnosis, disease prognosis evaluation, vaccine research and development, etc. Among them, ELISA is a highly sensitive in vitro antigen-antibody binding reaction detection method, which can be used for the detection of various pathogenic microorganisms and autoantigens. Compared with detection methods such as LC-MS / MS, ELISA detection has the advantages of simple operation, rapidity, simple experimental requirements, and convenient wide application.

[0006] However, there is currently a lack of relevant research and reports on immunological assays for escitalopram. To achieve highly sensitive and specific detection, the construction of antigens and antibodies used in immunological assays is a particularly crucial key point. Therefore, developing antigens and antibodies for escitalopram to achieve highly sensitive and specific immunological detection of escitalopram is an urgent problem to be solved in this field. Summary of the Invention

[0007] In view of the problems of the prior art, the present invention provides an escitalopram detection kit and a detection method.

[0008] An escitalopram detection kit, comprising:

[0009] 1) An R1 reagent containing an escitalopram antigen;

[0010] 2) An M1 reagent containing an escitalopram antibody;

[0011] The structure of the escitalopram antigen is as follows:

[0012]

[0013] n is selected from 2 - 10;

[0014] L is none or a linker;

[0015] R1 is selected from carrier proteins;

[0016] The escitalopram antibody is an antibody produced corresponding to the escitalopram antigen.

[0017] Preferably, n is 2, and the linker is

[0018] Preferably, the carrier protein is selected from keyhole limpet hemocyanin, ovalbumin, or bovine serum albumin.

[0019] Preferably, the R1 reagent comprises an escitalopram antigen conjugated with a luminescent substance.

[0020] Preferably, the R1 reagent is prepared by the following method:

[0021] Step 1, mix the escitalopram antigen with a luminescent substance solution and react;

[0022] Step 2, add a termination solution to terminate the reaction;

[0023] Step 3, dilute the escitalopram antigen labeled with the luminescent substance with a buffer solution to obtain the R1 reagent.

[0024] Preferably, the escitalopram antibody is a monoclonal antibody or a polyclonal antibody.

[0025] Preferably, the M1 reagent comprises an escitalopram antibody conjugated with a fluorescent label.

[0026] Preferably, the M1 reagent is prepared by the following method:

[0027] Step a: React the citalopram antibody with a biotin ester, and purify to obtain a biotinylated escitalopram antibody;

[0028] Step b: React the biotinylated escitalopram antibody with a fluorescent label to obtain an escitalopram antibody labeled with the fluorescent label;

[0029] Step c: Add a blocking solution, wash, and suspend with a buffer to obtain the M1 reagent.

[0030] The present invention also provides a method for detecting the content of escitalopram, and the method is to detect the content of escitalopram in a sample to be detected by using the above kit.

[0031] Preferably, the method comprises the following steps:

[0032] (i) Perform chemiluminescence detection by using an escitalopram calibrator, reagent R1 and reagent M1 to obtain a detection signal value, and draw a standard curve;

[0033] (ii) Perform chemiluminescence detection by using the sample to be detected, reagent R1 and reagent M1 to obtain a detection signal value, and convert the detection signal value into the content of escitalopram in the sample to be detected according to the standard curve.

[0034] In the prior art, the immunological detection reagent for escitalopram can only use rabbit polyclonal antibodies, and the specificity of polyclonal antibodies may be difficult to meet the performance requirements of the kit. The present invention constructs a detection kit comprising an escitalopram antigen and an antibody, and the immunological detection of escitalopram can be realized by using this kit. The present invention has the advantages of high sensitivity, good specificity, good accuracy and repeatability, and can meet the requirements of clinical detection. In addition, the kit and detection method of the present invention also have the advantages of good repeatability, simple operation, more in line with the requirements of clinical promotion, and have good application prospects.

[0035] Obviously, according to the above content of the present invention, other various forms of modifications, substitutions or changes can be made without departing from the above basic technical idea of the present invention according to the common general technical knowledge and conventional means in the art.

[0036] The following is a further detailed description of the above content of the present invention in the form of specific embodiments. 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. Description of the Drawings

[0037] Figure 1 1H NMR characterization results of the escitalopram hapten for Example 1;

[0038] Figure 2 Mass spectrometry characterization results of the escitalopram hapten for Example 1;

[0039] Figure 3 Escitalopram concentration detection results of 30 samples in Example 1. Specific Embodiments

[0040] In the following examples and experimental examples, the reagents and raw materials not specifically described are all commercially available products.

[0041] Example 1 Escitalopram Detection Kit and Escitalopram Detection Method

[0042] I. Escitalopram Detection Kit

[0043] The kit of this example is composed as follows (for 10 persons):

[0044] Table 1

[0045] Composition Specification M1 Reagent 0.5ml R1 Reagent 1ml

[0046] Among them, the structures and preparation methods of Reagent M1 and Reagent R1 are as follows:

[0047] (I) Escitalopram Antigen and Escitalopram Antibody

[0048] 1. Escitalopram Hapten

[0049] The structural formula of the escitalopram hapten in this example is as follows:

[0050]

[0051] Its preparation method is:

[0052]

[0053] S1. Compound 1 reacts with water to obtain Compound 2;

[0054] S2. Compound 2 and Compound 3 undergo a nucleophilic substitution reaction to obtain Compound 4;

[0055] S3. Compound 4 is hydrolyzed to obtain Compound 5.

[0056] Step S1 specifically includes: Mix ethanol and water in a volume ratio of 1:1. Take 20 ml of the mixed ethanol and water, and add Compound 1 (2.0 g, 6.17 mmol) to dissolve. Add NaOH (0.49 g, 12.34 mmol), and heat the reaction mixture at 80 °C for 16 h. The crude product of Compound 2 is dried over anhydrous Na2SO4 (vacuum can reduce the boiling point of the liquid and has the function of rapid drying). Dry it with a rotary evaporator to obtain a solid powder. Purify the dried Compound 2 by column chromatography to obtain the white solid, which is the product Compound 2 (1.27 g, yield 59.99%), and the specific mobile phase of the chromatography column is dichloromethane - methanol 15:1.

[0057] Step S2 specifically includes: Dissolve Compound 2 (500 mg, 1.46 mmol) in anhydrous DMF (10 mL), add Compound 3 (0.23 g, 1.21 mmol), add a mixed solution of T3P and ethyl acetate (T3P is mixed with 50% aqueous ethyl acetate solution in a volume ratio of 1:1, containing 1.39 g of T3P, 2.19 mmol) and DIEA (0.51 mL, 2.92 mmol), and stir at 25 °C for 1 h to obtain the crude product of Compound 4; Primary purification: Dilute the crude product of Compound 4 with water, and then subject the mixed solution to column chromatography for chromatographic separation to obtain the organic layer for standby (specifically, the chromatography column is a silica gel column, and the mobile phase in the chromatography column is dichloromethane); Secondary purification: Wash the organic layer 3 times with saturated NaHCO3 aqueous solution; Purify it by column chromatography (dichloromethane - methanol, 10:1) to obtain white oil, which is Compound 4 (50.01 mg, yield 16.1%).

[0058] Step S3 specifically includes: Dissolve Compound 4 (0.3 g, 0.64 mmol) in 6 ml of dichloromethane, add trifluoroacetic acid (3 mL, 38.515 mmol), and stir at room temperature for 1 h; Dry it with a rotary evaporator to obtain a solid powder; Dissolve the solid with ethyl acetate, and wash it with saturated NaHCO3 and saturated brine respectively; Dry it with anhydrous Na2SO4. After evaporation of the solvent, the residue is purified by column chromatography (dichloromethane:methanol = 10:1) to obtain Compound 7, a white solid, with a yield of 42.3%.

[0059] Product characterization (1H NMR data is as Figure 1 shown, and the mass spectrometry data is as Figure 2As shown: 1H NMR (400 MHz, CD3OD) δ 7.77 (d, J = 8.0 Hz, 1H), 7.70 (s, 1H), 7.60 - 7.50 (m, 3H), 7.06 (t, J = 8.8 Hz, 2H), 5.21 (q, J = 12.8 Hz, 2H), 3.61 (t, J = 6.8 Hz, 2H), 3.13 (t, J = 8.2 Hz, 2H), 2.79 (s, 6H), 2.61 (t, J = 6.8 Hz, 2H), 2.33 (ddd, J = 14.8, 9.6, 5.4 Hz, 1H), 2.26 - 2.16 (m, 1H), 1.74 - 1.58 (m, 2H). LC-MS (ESI+) m / z 415.2 (M + H)+.

[0060] 2. Escitalopram antigen

[0061] The structure is as follows:

[0062]

[0063] Among them, L is a linker, and the linker is R1 is a carrier protein. The carrier protein is selected from keyhole limpet hemocyanin, ovalbumin or bovine serum albumin. In this example, bovine serum albumin and ovalbumin are taken as examples to illustrate its preparation method:

[0064] S1. Activation of the hapten:

[0065] Take the escitalopram hapten and dissolve it in dimethylformamide solution to prepare an 800 mL escitalopram antigen solution with a concentration of 5×10 -5 mg / μL. Then add 5 mg EDC and 5 mg NHS, and react at 25°C in the dark for 1 - 6 hours to obtain a reaction solution, and the solution is colorless, clear and transparent;

[0066] S2. Small molecule labeling;

[0067] Weigh 10 mg of bovine serum albumin (or 10 mg of ovalbumin) and dissolve it in 5 ml of 0.01 M PBS (pH 7.4) solution, which is colorless, clear and transparent; mix the reaction solution in step S1 with the bovine serum albumin (or ovalbumin) solution to obtain a slightly milky turbid solution, and react at 25°C in the dark for 8 h, and the solution is still slightly milky turbid;

[0068] S3. Centrifuge the turbid solution (6000 r / min, 5 min) and extract the supernatant;

[0069] S4. 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;

[0070] S5. Centrifuge the retained solution after dialysis (6000 r / min, 5 min), and extract the supernatant to obtain the escitalopram-carrier protein complex, namely the escitalopram antigen.

[0071] 3. Escitalopram antibody

[0072] Use the above-mentioned escitalopram antigen for animal immunization to prepare paired monoclonal antibodies, which specifically include the following steps:

[0073] Step a. After immunizing the animal (mice are selected in this example) multiple times by the conventional Freund's adjuvant method with the escitalopram antigen, obtain the serum containing specific antibodies. After the titer test is qualified, take the spleen cells for fusion;

[0074] Step b. Screen the positive fusion cells and perform subcloning to obtain monoclonal cell lines;

[0075] Step c. Produce ascites or produce by cell culture, separate and purify to obtain monoclonal escitalopram antibody.

[0076] Use the ELISA method to test the titer of the prepared escitalopram antibody. The specific detection method is as follows:

[0077] 1.1 Reagents

[0078] 1.1.1 Escitalopram antibody (produced using the escitalopram antigen with bovine serum albumin as the carrier protein) and escitalopram antigen (ovalbumin as the carrier protein);

[0079] 1.1.2 HRP-labeled goat anti-mouse enzyme-labeled secondary antibody;

[0080] 1.1.3 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);

[0081] 1.1.4 0.1M PH = 9.6 sodium bicarbonate buffer solution;

[0082] 1.1.5 0.01M PBS;

[0083] 1.1.6 Termination solution (2M H2SO4 solution or 1M HCl solution);

[0084] 1.1.7 Standard solution;

[0085] 1.2 Method

[0086] 1.2.1 Indirect ELISA (detecting the titer of antigen and antibody)

[0087] a) Dilute escitalopram antigen to 1 μg / mL with 0.1 M sodium bicarbonate buffer at pH 9.6, add it to a 96-well enzyme-linked immunosorbent assay (ELISA) plate, 100 μL per well, and react at 37 °C for 3 h or leave it standing overnight at 4 °C.

[0088] b) Discard the liquid in the wells of the plate, add 250 μL of ELISA washing solution, leave it standing for 30 s, discard the liquid in the plate, and repeat 3 times. (Wash away the uncoated free antigen).

[0089] c) Dilute the escitalopram antibody to 1 mg / mL with 0.01 M phosphate-buffered saline (PBS), and then further dilute it to 1k, 2k, 4k, 8k, 16k, 32k, 64k with 0.01 M PBS in labeled centrifuge tubes for standby.

[0090] d) Add the diluted escitalopram antibody prepared in the above step (negative control is 0.01 M PBS), 100 μL per well, and make three replicates for each concentration gradient, and react at 37 °C for 45 min.

[0091] e) Discard the liquid in the wells of the plate, add 250 μL of ELISA washing solution, leave it standing for 30 s, discard the liquid in the plate, and repeat 3 times.

[0092] f) Dilute the horseradish peroxidase (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.

[0093] g) Discard the liquid in the wells of the plate, add 250 μL of ELISA washing solution, leave it standing for 30 s, discard the liquid in the plate, and repeat 3 times.

[0094] 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 for 15 min in the dark at room temperature (18 - 28 °C environment).

[0095] i) Add the stop solution (1 M HCl solution or 2 M H2SO4 solution), 100 μL per well, OD 450 Read the OD value.

[0096] Note: 1 M HCl solution is prepared from 6 M HCl solution, and it 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.

[0097] 1.2.2 Competitive ELISA (detecting antigen-antibody competition)

[0098] a) Dilute escitalopram 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 let it stand overnight at 4 °C.

[0099] b) Discard the liquid in the wells, 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)

[0100] c) Dilute the escitalopram antibody 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.

[0101] d) Add 50 μL of the diluted standard solution (negative control is 0.01M PBS) to each well, and then add 50 μL of the diluted escitalopram antibody to each well (negative control is 0.01M PBS). Do three replicates for each concentration gradient and react at 37 °C for 45 min.

[0102] e) Discard the liquid in the wells, add 250 μL of ELISA washing solution, let it stand for 30 s, discard the liquid in the plate, and repeat 3 times.

[0103] f) Dilute the HRP-labeled goat anti-mouse enzyme conjugate according to the instructions (dilute with 0.01M PBS), add the diluted HRP-labeled goat anti-mouse enzyme conjugate, 100 μL per well, and react at 37 °C for 45 min.

[0104] g) Discard the liquid in the wells, add 250 μL of ELISA washing solution, let it stand for 30 s, discard the liquid in the plate, and repeat 3 times.

[0105] 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.

[0106] i) Add the stop solution (1M HCl solution or 2M H2SO4 solution), 100 μL per well, OD 450 Read the OD value.

[0107] Note: The 1M HCl solution is prepared from 6M HCl solution and should be prepared freshly before use. 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 well.

[0108] The results are as follows:

[0109] Table 2 Results of the determination of the titer inhibition of the escitalopram antibody

[0110]

[0111] The prepared escitalopram antibody was subjected to IC50 testing. The results showed that the IC50 was 8.10 ng / mL, indicating that the escitalopram antibody had high sensitivity and good specificity.

[0112] (II) Reagent M1 and Reagent R1

[0113] 1. Biotin ester-labeled antibody

[0114] (1) Put 0.1 mg of the above-mentioned escitalopram antibody (produced using escitalopram antigen with bovine serum albumin as the carrier protein) into PBS buffer and dialyze for 24 - 48 hours, changing the buffer twice in the middle;

[0115] (2) Dilute the antibody recovered after dialysis to 0.2 mg / mL, add a certain amount of biotin ester, react at room temperature for 40 min, terminate the reaction with a termination solution (1 M lysine or 1 M Tris) for 30 min, and then transfer it into a dialysis bag and dialyze for 24 - 48 hours.

[0116] (3) Recover the dialyzed biotinylated antibody, add an equal volume of glycerol, label it and store it at -20 ± 5 °C.

[0117] 2. Preparation of Reagent M1

[0118] (1) Take 100 mg of magnetic particles and wash them 2 - 5 times with Tris-HCL buffer;

[0119] (2) Add 0.1 mg of biotinylated antibody at a ratio of 1 μg / mg and mix well at room temperature for 40 min;

[0120] (3) Add 1 mL of magnetic particle blocking solution at a ratio of 10 μL / mg. The blocking solution is a 3 g / L vitamin H solution, and react at room temperature for 40 min;

[0121] (4) Wash 2 - 5 times with Tris-HCL buffer, suspend, and the final concentration is 0.4 mg / mL.

[0122] 3. Acridinium ester-labeled escitalopram antigen

[0123] (1) Take 0.1 mg of the above-mentioned escitalopram antigen (with ovalbumin as the carrier protein), dilute the concentration to 1 mg / mL, add 4 μL of 5 mmol / l acridinium ester, react in the dark at room temperature for 2 hours, and then add 1 M lysine or 1 M Tris to terminate the reaction for 30 min.

[0124] (2) Filter and purify the acridine-labeled product with a filter column, add an equal volume of glycerol, label it and store it at -20 ± 5 °C.

[0125] 4. Preparation of Reagent R1:

[0126] Dilute the acridine-labeled escitalopram antigen mentioned above with PBS buffer at a volume ratio of 1:2000.

[0127] II. Escitalopram Detection Method

[0128] The method for detecting the concentration of escitalopram in a blood sample using the above-mentioned kit is as follows:

[0129] (1) Mix the blood sample to be tested, Reagent M1, and Reagent R1 at a volume ratio of 1:5:10;

[0130] (2) Perform luminescence detection on the mixed solution in step (1) to obtain a detection signal value;

[0131] (3) Calculate the content of escitalopram in the sample to be tested by converting the signal value according to the standard curve.

[0132] Among them, the standard curve is drawn as follows: Use the escitalopram calibrator, Reagent R1, and Reagent M1 for chemiluminescence detection to obtain the detection signal value and draw the standard curve. In this embodiment, the data points for drawing the standard curve are as follows:

[0133] Table 3 Data Points of the Standard Curve

[0134] Calibrator Concentration (ng / mL) RLU 0 1180620 4 912444 8 712781 20 402498 50 188780 200 45760

[0135] Thirty samples were respectively detected using this embodiment and the existing LC / MS clinical method, and the results are compared in Table 4 and Figure 3 as shown.

[0136] Table 4 Detection Results of Escitalopram Concentration in 30 Samples

[0137]

[0138]

[0139] From the above data, it can be seen that the escitalopram concentration results obtained by the detection method of this embodiment are close to those obtained by the existing LC / MS clinical method, indicating that the detection method of this embodiment has good accuracy.

[0140] Repeatability experiments were carried out on standard samples with escitalopram concentrations of 10 ng / mL and 100 ng / mL, and the results are shown in the following table:

[0141] Table 5 Results of Repeatability Experiments

[0142]

[0143] It can be seen that for samples with different concentrations, the CV values of the concentration detection results of 10 repeated experiments are 1.23%-2.52%, indicating that the detection method of this embodiment has good repeatability and can meet the requirements of clinical detection.

[0144] As can be seen from the above embodiments, the present invention constructs a new escitalopram detection kit and detection method, which have good sensitivity and specificity for blood escitalopram. Moreover, the detection method of the present invention has good accuracy and repeatability and can meet the requirements of clinical detection. Therefore, the present invention has good application prospects.

Claims

1. An escitalopram detection kit, characterized in that it Comprising: 1) An R1 reagent containing an escitalopram antigen; 2) An M1 reagent containing an escitalopram antibody; The structure of the escitalopram antigen is as follows: n is selected from 2 - 10; L is none or a linker; R1 is selected from carrier proteins; The escitalopram antibody is an antibody produced corresponding to the escitalopram antigen.

2. The kit according to claim 1, wherein The value of n is 2, and the linker is 3. The kit according to claim 1, characterized in that: The carrier protein is selected from keyhole limpet hemocyanin, ovalbumin or bovine serum albumin.

4. The kit according to claim 1, characterized in that: The R1 reagent comprises an escitalopram antigen conjugated with a luminescent substance.

5. The kit according to claim 4, wherein: The R1 reagent is prepared according to the following method: Step 1, mix the escitalopram antigen with a luminescent substance solution and react; Step 2, add a termination solution to terminate the reaction; Step 3, dilute the escitalopram antigen labeled with the luminescent substance with a buffer solution to obtain the R1 reagent.

6. The kit according to claim 1, characterized in that: The escitalopram antibody is a monoclonal antibody or a polyclonal antibody.

7. The kit according to claim 1, characterized in that: The M1 reagent comprises an escitalopram antibody conjugated with a fluorescent label.

8. The kit according to claim 7, characterized in that: The M1 reagent is prepared according to the following method: Step a, react the escitalopram antibody with a biotin ester to obtain a biotinylated escitalopram antibody; Step b, react the biotinylated escitalopram antibody with a fluorescent label to obtain an escitalopram antibody labeled with the fluorescent label; Step c, add a blocking solution, wash, and suspend with a buffer solution to obtain the M1 reagent.

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

10. The method according to claim 9, wherein The method comprises the following steps: (i) Perform chemiluminescence detection using an escitalopram calibrator, reagent R1 and reagent M1 to obtain a detection signal value and plot a standard curve; (ii) Perform chemiluminescence detection using the sample to be detected, reagent R1 and reagent M1 to obtain a detection signal value, and convert the detection signal value into the content of escitalopram in the sample to be detected according to the standard curve.

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