CN115267195A - A thymidine kinase 1 magnetic particle chemiluminescence detection reagent - Google Patents

Abstract

The invention belongs to the technical field of biological detection, and particularly relates to a thymidine kinase 1 magnetic particle chemiluminescence detection reagent which comprises a magnetic particle suspension coated with an anti-TK 1 antibody, a detection reagent of an acridinium ester labeled anti-TK 1 detection antibody, a TK1 calibrator and a TK1 quality control product, wherein the magnetic particle suspension coated with the anti-TK 1 antibody is prepared from reagent components, deionized water and magnetic particles coated with the TK1 antibody, the detection reagent of the acridinium ester labeled anti-TK 1 detection antibody is the acridinium ester labeled anti-TK 1 antibody, and the detection reagent of the acridinium ester labeled anti-TK 1 detection antibody is stored in a luminophore diluent.

Description

A kind of thymidine kinase 1 magnetic particle chemiluminescent detection reagent

technical field

The invention relates to the technical field of biological detection, in particular to a thymidine kinase 1 magnetic particle chemiluminescent detection reagent.

Background technique

Thymidine kinase 1 is one of the key enzymes in the pyrimidine salvage synthesis pathway in the cell proliferation cycle. It is a new specific marker for cell proliferation and can be used to evaluate the cell division and proliferation cycle, early screening of tumors, and assessment of tumor staging , The role of treatment effect monitoring and prognosis evaluation has attracted increasing attention. Usually, the average level of TK1 in normal people is about 1pM. The specific value depends on individual differences, but in general it is between 0-2pM. For abnormal proliferation lesions , due to the loss of apoptosis regulation in diseased cells, the sharp increase in DNA synthesis will lead to an abnormal increase in TK1 levels, that is, the serum TK1 enzyme content in patients with malignant proliferation lesions will be 2-200 times higher than normal people, so TK1 is currently It is also used as a non-specific tumor marker, which is applied to the screening of malignant proliferation lesions in physical examination and clinic and the follow-up monitoring after treatment. The detection of TK1 level has application value in the monitoring of malignant solid tumors such as breast cancer, lung cancer and gastric cancer. Higher, as a preferred marker for assessing the proliferation rate of tumor cells, TK1 is minimal or undetectable in the serum of healthy people, but it increases with the rapid proliferation of tumor cells in patients with malignant tumors. Therefore, it is established An efficient, sensitive, simple and quick TK1 detection technology has very important practical significance.

At present, the commonly used method for determining the content of TK1 in samples is still based on enzyme-linked immunosorbent assay. This method is still mainly operated manually or semi-automatically, which is time-consuming and laborious, and has a great impact on the results. At the same time, its detection sensitivity and linear range It is still relatively low, and it is still mainly used for laboratory analysis. It has great limitations in clinical detection applications, low efficiency, and still cannot meet the growing needs of clinical practice. Chemiluminescence immunoassay is a combination of chemiluminescence technology and immunoassay A new analytical method combining analytical methods, which has the advantages of high sensitivity, strong specificity, simple and quick operation, etc., has been widely used in many detection fields. At present, there are still few reports on TK1 magnetic particle chemiluminescence detection method , the present invention uses the TK1 antibody-magnetic particle conjugate to bind and separate the TK1 protein, and the TK1 paired antibody-acridine ester luminescent agent emits light, and establishes a TK1 magnetic particle chemiluminescent detection method, which is highly sensitive and specific for TK1 , Accurate detection provides a new automatic means of a new technology.

Therefore, we propose a thymidine kinase 1 magnetic particle chemiluminescent detection reagent.

Contents of the invention

In view of the above and/or problems existing in a thymidine kinase 1 magnetic particle chemiluminescence detection reagent, the present invention is proposed.

Therefore, the object of the present invention is to provide a kind of thymidine kinase 1 magnetic particle chemiluminescent detection reagent, by chemiluminescent method, immune sandwich principle, specimen, standard product and mouse monoclonal antibody labeled with acridinium ester, coated with another strain The magnetic microspheres of the mouse monoclonal antibody are mixed to form an immune complex of another monoclonal antibody, which can solve the above-mentioned existing problems.

In order to solve the above technical problems, according to one aspect of the present invention, the present invention provides the following technical solutions:

A thymidine kinase 1 magnetic particle chemiluminescence detection reagent, which includes: magnetic particle suspension coated with anti-TK1 antibody, acridinium ester-labeled anti-TK1 detection antibody detection reagent, TK1 calibrator and TK1 quality control product;

The magnetic particle suspension coated with anti-TK1 antibody is prepared from reagent components, deionized water and magnetic particles coated with TK1 antibody;

The detection reagent of the acridinium ester-labeled anti-TK1 detection antibody is an acridinium ester-labeled anti-TK1 antibody, and the detection reagent of the acridinium ester-labeled anti-TK1 detection antibody is stored in the diluent of the luminescent substance.

As a preferred scheme of a thymidine kinase 1 magnetic particle chemiluminescent detection reagent according to the present invention, wherein: the magnetic particle suspension coated with anti-TK1 antibody is made of 0.05-0.15mol/L Tris- Preparation of HCl buffer, 0.5%-1.5% sodium chloride, 0.025%-10.075% surfactant, 1.5%-2.5% bovine serum albumin, 3%-5% trehalose and magnetic microsphere-labeled TK1 antibody conjugate made.

As a preferred scheme of a thymidine kinase 1 magnetic particle chemiluminescence detection reagent of the present invention, wherein: the detection reagent of the acridinium ester-labeled anti-TK1 detection antibody is made of 0.01mol/L phosphate buffer Liquid, 0.25%-0.75% casein, 0.025%-10.075% surfactant, 3%-5% trehalose and acridinium ester labeled anti-TK1 detection antibody.

As a preferred scheme of a thymidine kinase 1 magnetic particle chemiluminescent detection reagent according to the present invention, wherein: the diluent of the luminescent substance is set as deionized water: 100mL, Na ₂ HPO ₄ : 0.54g, NaH ₂ PO ₄ 2H ₂ O: 0.967g, NaCl: 0.90g, Casein: 0.25%-0.75%, Trehalose: 3%-5%, Proclin 300: 0.1%-0.5%, and Triton X-100: 0.025%- 0.075%.

As a preferred scheme of a thymidine kinase 1 magnetic particle chemiluminescent detection reagent of the present invention, wherein: the TK1 calibrator and the TK1 quality control are both made of 0.01mol/L phosphate buffer, 0.25 %-0.75% casein and recombinant human TK1 protein formulated.

As a preferred scheme of a thymidine kinase 1 magnetic particle chemiluminescence detection reagent according to the present invention, wherein: the calibration product diluent and the sample diluent are formulated into: KCl: 0.30 g, NaCl: 12.00 g, KH ₂ PO ₄ : 0.30g, Na ₂ HPO ₄ : 1.725g, PH: 7.2-7.6, casein: 0.25%-0.75%, deionized water: 1500ml.

A preparation method of thymidine kinase 1 magnetic particle chemiluminescence detection kit, comprising the following steps:

Step 1: Preparation of magnetic particle suspension coated with anti-TK1 antibody:

Add Tris-HCl buffer salt, sodium chloride, surfactant, trehalose, bovine serum albumin, and preservatives in deionized water in sequence. Each time you add materials, you need to stir until they are completely dissolved, adjust the pH value to 6.0-8.5, and mix After uniformity, pass through a 0.22 μm filter membrane to obtain the magnetic microsphere dilution, and then add the magnetic microsphere-labeled TK1 antibody conjugate to obtain the magnetic particle suspension coated with anti-TK1 antibody;

Step 2: Preparation of detection reagents for acridinium ester-labeled anti-TK1 detection antibodies:

Add phosphate buffer salt, surfactant, casein, trehalose, and preservatives in deionized water in sequence. Each addition needs to be stirred until completely dissolved, and the pH value is adjusted to 6.0-8.5. After mixing evenly, pass through a 0.22 μm filter membrane Obtain the diluent of the luminescent substance, and then add the acridinium ester-labeled anti-TK1 detection antibody conjugate to obtain the detection reagent of the acridinium ester-labeled anti-TK1 detection antibody;

Step 3: Preparation of TK1 calibrator and TK1 quality control

Add phosphate buffer salt, casein, and preservatives in deionized water in sequence. Each time you add materials, you need to stir until they are completely dissolved. Adjust the pH value to 6.0-8.5. Add the recombinant human TK1 protein to obtain the corresponding calibrator series and quality control series;

Step 4: Assembling the kit: the above prepared magnetic particle suspension coated with anti-TK1 antibody and the detection reagent of anti-TK1 detection antibody labeled with acridinium ester in volume ratio R1:R2=1:1, and the corresponding calibration Product and quality control product series, the volume ratio is 1:1, divided into bottles to form a kit.

Compared with existing technology:

The kit of the present invention adopts the chemiluminescent immunoassay method of acridinium ester luminescence, which has the advantages of high signal-to-noise ratio and high cost-effectiveness, and meanwhile the specificity and sensitivity of the method have also been greatly improved, which is a rapid detection method for clinical diagnosis. Provides excellent support;

Two kinds of markers are provided in the kit of the present invention: one is magnetic microspheres and anti-TK1 antibody coupling magnetic bead suspension, and the other is acridinium ester and anti-TK1 antibody coupling detection reagent, using double antibody The sandwich method, with the help of automated operating equipment, can quickly, accurately and sensitively detect the content of TK1 in the sample to be tested;

The kit of the present invention is operated by means of a fully automatic chemiluminescence immunoassay analyzer, and all samples and reagents are added by the instrument, which reduces labor costs, reduces the interference of manual operations on test results, and also greatly shortens the test time. Provide test results more efficiently and quickly;

The kit of the present invention does not need to dilute the sample to be tested, and can be directly tested on a machine, thereby reducing errors caused by dilution and improving the detection rate of low-value samples.

Description of drawings

Fig. 1 is the calibration curve diagram of the reagent prepared by the present invention on the running Cosman 500s chemiluminescence detector;

Fig. 2 is the TK1 dilution linear detection data diagram provided by the present invention;

Fig. 3 is the detection result figure of TK1 magnetic particle chemiluminescence detection reagent spiked recovery rate provided by the present invention;

Figure 4 is an analysis diagram of the precision test results of the TK1 magnetic particle chemiluminescence reagent provided by the present invention;

Fig. 5 is a test result diagram of the influence test of the interfering reagent provided by the present invention on the detection of the TK1 magnetic particle chemiluminescent reagent;

Figure 6 is a diagram of the specific analysis results of the TK1 detection kit provided by the present invention;

Fig. 7 is a diagram showing the correlation analysis between the test results provided by the present invention and the test results of the comparison reagents.

Detailed ways

In order to make the purpose, technical solution and advantages of the present invention clearer, the following will further describe the implementation of the present invention in detail in conjunction with the accompanying drawings.

The present invention provides a thymidine kinase 1 magnetic particle chemiluminescent detection reagent, please refer to Figure 1-7, comprising the following steps:

Step 1: Preparation of magnetic particle suspension coated with anti-TK1 antibody:

In this invention, the process of connecting the amino groups on the anti-TK1 antibody and the carboxylated magnetic microspheres to the magnetic beads by means of covalent coupling is called the preparation of the magnetic particle suspension coated with the anti-TK1 antibody; preparation The obtained magnetic particle suspension of anti-TK1 antibody is called magnetic particle coating. The basic principle of the coating is that the active amino groups on the surface of the antibody and the carboxyl chemical groups on the surface of the magnetic microspheres, under the action of a chemical cross-linking agent, , a chemical reaction occurs to form a covalent conjugate of antibody-magnetic microspheres. The covalent conjugate is washed and blocked to remove unreacted antigen or antibody, and to block non-specific binding sites, and finally prepared into a kit The magnetic particle suspension of the anti-TK1 antibody, select the appropriate anti-TK1 antibody and carboxylated magnetic microspheres for coupling,

The specific steps are as follows: carboxyl magnetic particles are activated by EDC/sulfo-NHS method, and then coupled with anti-TK1 antibody;

The specific steps are: wash carboxyl magnetic particles with magnetic particle buffer (0.05M morpholine sulfonic acid, pH 6.0) for 3 times (200uL/time), add the same amount of EDC and sulfo-NHS, shake and activate at 37°C for 25min ;

After washing twice with magnetic particle buffer (200uL/time) with the help of a magnetic stand, add anti-TK1 antibody (diluted to 0.5mg/mL with magnetic particle buffer), the mass ratio of antibody to magnetic beads is 1:10, and shake at room temperature Coupling 5h;

Then use blocking buffer (0.05M Tris, 0.5% casein, 0.05% Triton X-100, 0.5% Proclin300) to block the magnetic particles at 37°C for 1.5h, and wash the magnetic particle buffer twice (200uL/time);

Finally, the TK1 antibody-magnetic particle conjugate was diluted to 0.5 mg with magnetic particle diluent (0.05M Tris, 1% sodium chloride, 2% BSA, 4% trehalose, 0.05% Triton X-100, 0.5% Proclin300) /mL (antibody concentration), stored in a refrigerator at 4°C;

Step 2: Preparation of detection reagents for acridinium ester-labeled anti-TK1 detection antibodies:

Select TK1 paired antibody to couple with acridinium ester to prepare TK1 paired antibody-acridine ester luminescent agent;

Specific steps: according to a certain mass ratio, mix the TK1 paired antibody with acridinium ester, shake and react at 37°C for 30 minutes, and add 0.5% lysine at a final concentration to terminate the reaction;

Dialysis against 0.1M PBS to remove unbound acridinium esters;

After purification by Sephadex G50 gel chromatography, the product with high luminescence intensity was collected. Dilute the purified TK1 paired antibody-acridinium ester luminescent agent with the luminescent substance diluent to the working concentration, store it in a refrigerator at 4°C for later use, and obtain the detection reagent of the acridinium ester-labeled anti-TK1 detection antibody;

Kit composition, as shown in the table below

Step 3: Preparation of TK1 calibrator and TK1 quality control

Add phosphate buffer salt, casein, and preservatives in deionized water in sequence. Each time you add materials, you need to stir until they are completely dissolved. Adjust the pH value to 6.0-8.5. Add the recombinant human TK1 protein to obtain the corresponding calibrator series and quality control series;

Step 4: Assembling the kit: the above prepared magnetic particle suspension coated with anti-TK1 antibody and the detection reagent of anti-TK1 detection antibody labeled with acridinium ester in volume ratio R1:R2=1:1, and the corresponding calibration Product and quality control product series, the volume ratio is 1:1, divided into bottles to form a kit;

Evaluation of kit performance:

The magnetic particle suspension coated with anti-TK1 antibody prepared as above, the detection reagent of anti-TK1 detection antibody labeled with acridinium ester, calibrator 1, calibrator 2, quality control product 1, quality control product 2, and master curve The cards are individually packaged, and then assembled into a finished kit. The detection steps of the TK1 magnetic particle chemiluminescence detection kit are completed by Cosman 500 equipment (supported with washing solution PBST (0.05% tween20 dissolved in 10mM PBS) and luminescence Substrate solutions A and B are as follows:

Take 50 μL of TK1 antibody-magnetic particles (20 μg/mL), add 50 μL of TK1 standard or serum sample and 50 μL of TK1 paired antibody-acridinium ester luminescent agent, incubate at 37°C for 20 minutes, and the magnet will attract the magnetic particles and their magnetic particle pairs. Combine the material to the bottom of the test tube, wash the test tube gently with washing buffer (PBS+0.05% Tween20) to remove free substances, then add 100 μL of luminescent substrate to detect the luminescence value, and the above detection steps are all carried out in a chemiluminescence detector. Automatic detection without manual operation, the luminescence value is positively correlated with the concentration of TK1 in the sample;

Evaluation of kit methodology:

a. Standard curve drawing

The traceability program of the TK1 detection kit designed in this research adopts chemiluminescence immunoassay method. Currently, there is no national standard for TK1, so the calibrator can be traced to the diluent of calibrator (0.01M PBS, 2% trehalose, 0.2% casein, 0.5% histidine, 0.5% Proclin300) to dilute the TK1 antigen, traced to the primary calibrator of the comparison kit (certified luminescent reagent), the deviation is controlled within ±10%. Through repeated experiments, the standard curve is drawn as follows: prepare a series of concentrations of recombinant TK1 protein standards (0, 1.25, 2.5, 5, 10, 20, 40 pmol/L), and use this method to detect the luminescence corresponding to each standard concentration Value, draw standard curve, each concentration measures 3 duplicate wells, adopt double logarithm mathematical model (Log-Logit) to carry out the fitting of standard curve, the results are shown in Fig. 1;

b. Dilution linear detection

Dilute the high-value samples close to the upper limit of the linear range of the calibration curve for no less than 5 gradients to the lower limit of the linearity of the method, repeat the measurement for each concentration three times, and use the least square method to linearly fit the measured results and the corresponding dilution ratios , and calculate the interrelationship between the two to obtain the linear correlation coefficient R, the results are shown in Figure 2;

c. Sensitivity assessment

Prepare five clinical samples or healthy human serum samples with a value close to 0, and use the reagents in this study for detection. Each sample is repeated 3 times and continuously detected for 4 days to obtain the mean and standard deviation of luminescence of all samples; prepare five concentration ranges Serum samples between 1-4 times the LoB were tested four times a day with an interval of more than 2hrs, and each test was repeated 3 times for a total of five days. Data processing and analysis were performed according to the methods and formulas in the EP17-A2 file, and calculation The limit of blank (LoB), limit of detection (LoD), and functional sensitivity (FS) are obtained. The final results of the sensitivity test evaluation of this method are: LOB: 0.70pmol/L, LOD: 1.02pmol/L, FS: 1.50pmol/L .

c. Accuracy evaluation

Evaluate the spiked recovery rate of the detection reagent: add the TK1 protein standard of known concentration to the matrix serum, and the volume of the high-value sample added does not exceed 10% of the total volume, and prepare the serum with the spiked concentration Samples were tested 3 times using this method, and the mean values, SD, coefficient of variation (CV), and recovery of standard additions were calculated for the three samples of high concentration samples, spiked concentration samples, and matrix concentration samples, so as to evaluate the accuracy of the method. accuracy and repeatability. According to the formula: recovery rate of standard addition=(determined concentration-matrix concentration)/concentration of standard addition*100) to calculate the recovery rate of standard addition, the results are shown in Figure 3;

d. Precision (repeatability) evaluation

Use TK1 reagents to detect TK1 calibrator at high, medium and low concentration levels. Samples with different concentrations are tested twice a day in the morning and afternoon. Each test has 2 repetitions. The time interval is not less than 2h, and the continuous measurement is 20d. , calculate the intra-analysis precision, inter-analysis precision and total imprecision of each concentration sample respectively, the results are shown in Figure 4;

e. Interference experiment

In this evaluation, three common serum interfering substances were used, namely hemoglobin, triglyceride and bilirubin, and a certain concentration of interfering substances were added to the serum test samples with high concentration of TK1 and low concentration of TK1 respectively, as the interference For the substance sample, the volume of the interfering substance added does not exceed 5% of the total volume. The control group also adds the same volume of PBS as the control sample, and then the two groups of samples are tested separately, and each concentration is detected 3 times, and according to the formula: Interference Rate = (measured concentration of interfering substance samples - measured concentration of control group samples)/measured concentration of control group samples * 100%, the interference rate is calculated, and the results are shown in Figure 5;

f. Specificity assessment

This method simultaneously detects TK1 standard substance (30pmol/L), human serum albumin (100ng/mL), fibrinogen (100ng/mL), carcinoembryonic antigen (CEA), interleukin 6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) to evaluate the specificity of this method, the above-mentioned several proteins are common proteins in human serum, the results are shown in Figure 6;

Six calibrators and 100 serum samples were tested, and compared with the clinical test results of thymidine kinase 1 (TK1) assay kit (chemiluminescence immunoassay).

Experimental method: Cosman S500 luminescent fully automated instrument automatically absorbs 50ul of magnetic particle reagent (Ra), 50ul of acridinium ester reagent (Rb) and 50ul of samples into the reaction cup, incubates at 37°C for 20min, washes and pumps the luminescent substrate Solution A and B, and then the luminometer automatically collects the relative luminescence units (RLU) of the reaction solution in the cuvette. Make a calibration curve based on the concentration of RLU and calibrator 1 and 2, and then calculate the content of TK1 in the sample to be tested according to the curve after taking the luminescence value of the sample to be tested;

Linear range: 0-40pmol/L

Test instrument: Cosman S500.

After the reagents and luminescent reagents of this example were calibrated and tested for quality control products, they all reached the standard, and then tested 100 cases of clinical samples with different concentrations. The test results are shown in Table 1, and the correlation curve is shown in Figure 7:

Table 1. Comparison of clinically confirmed samples with the test results of this kit

Wherein the activation of magnetic microspheres adopts 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride solution and N-hydroxyl sulfosuccinimide, which improves the efficiency of Activation performance and efficiency;

Wherein the acidic buffer is selected from one or more of 2-(N-morpholine) ethanesulfonic acid buffer, borate buffer, phosphate buffer;

The pH value of the buffer solution is 5.8-6.2, the preservative is selected from proclin300; the surfactant is selected from Tween 20 or polyethylene glycol octylphenyl ether, and the magnetic particles coated with anti-TK1 antibodies are selected for different particle sizes and The complex formed by magnetic particles with surface active functional groups and TK1 antibody, the active functional groups on the surface of the magnetic particles are selected from p-toluenesulfonyl or carboxyl groups, and the particle diameter of the magnetic particles is 0.1-10 μm.

Among them, the magnetic particle suspension coated with anti-TK1 antibody, the TK1 antibody in the detection reagent of the anti-TK1 detection antibody labeled with acridinium ester are two kinds of monoclonal antibodies with different binding sites, and the two TK1 antibodies are selected from mouse anti-monoclonal antibodies. Cloned antibodies.

The volume ratio of the magnetic particle suspension coated with anti-TK1 antibody and the detection reagent of anti-TK1 detection antibody labeled with acridinium ester is 1:1-1:3;

The kit adopts the chemiluminescence immunoassay method of acridinium ester luminescence, which has the advantages of high signal-to-noise ratio and high cost performance. At the same time, the specificity and sensitivity of the method have also been greatly improved, providing a rapid detection for clinical diagnosis. As a good support, two markers are provided in the kit: one is the magnetic bead suspension of magnetic microspheres and anti-TK1 antibody, and the other is the coupling detection of acridinium ester and anti-TK1 antibody The reagent adopts the double-antibody sandwich method, and with the help of automated operating equipment, it can quickly, accurately and sensitively detect the content of TK1 in the sample to be tested. Through the kit, it is run by means of a fully automatic chemiluminescence immunoassay analyzer. All samples and reagents The addition of samples is performed by the instrument, which reduces labor costs, reduces the interference of manual operations on the test results, and greatly shortens the test time to provide test results more efficiently and quickly. When testing samples through the kit, no treatment is required. The test sample can be diluted, which can be directly tested on the machine, reducing the error caused by dilution and helping to improve the detection rate of low-value samples.

While the invention has been described above with reference to the embodiments, various modifications may be made and equivalents may be substituted for elements thereof without departing from the scope of the invention. In particular, as long as there is no structural conflict, the various features in the embodiments disclosed in the present invention can be used in combination with each other in any way, and the description of these combinations is not exhaustive in this specification only to show In consideration of omitting space and saving resources. Therefore, the present invention is not limited to the specific embodiments disclosed herein, but includes all technical solutions falling within the scope of the claims.

Claims (7)

1. A thymidine kinase 1 magnetic particle chemiluminescent detection reagent, characterized in that: comprising the magnetic particle suspension coated with anti-TK1 antibody, the detection reagent of the anti-TK1 detection antibody labeled with acridinium ester, TK1 calibrator and TK1 quality control; The magnetic particle suspension coated with anti-TK1 antibody is prepared from reagent components, deionized water and magnetic particles coated with TK1 antibody; The detection reagent of the acridinium ester-labeled anti-TK1 detection antibody is an acridinium ester-labeled anti-TK1 antibody, and the detection reagent of the acridinium ester-labeled anti-TK1 detection antibody is stored in the diluent of the luminescent substance. 2. a kind of thymidine kinase 1 magnetic particle chemiluminescent detection reagent according to claim 1, is characterized in that, the magnetic particle suspension that is coated with anti-TK1 antibody is made of 0.05-0.15mol/L Tris- Preparation of HCl buffer, 0.5%-1.5% sodium chloride, 0.025%-10.075% surfactant, 1.5%-2.5% bovine serum albumin, 3%-5% trehalose and magnetic microsphere-labeled TK1 antibody conjugate made. 3. a kind of thymidine kinase 1 magnetic particle chemiluminescence detection reagent according to claim 1, is characterized in that, the detection reagent of the anti-TK1 detection antibody of described acridinium ester is by the phosphate buffered saline of 0.01mol/L Liquid, 0.25%-0.75% casein, 0.025%-10.075% surfactant, 3%-5% trehalose and acridinium ester labeled anti-TK1 detection antibody. 4. A thymidine kinase 1 magnetic particle chemiluminescent detection reagent according to claim 1, characterized in that, the diluent of the luminescent substance is set as deionized water: 100mL, _{Na2HPO4 :} 0.54g, _NaH2 PO ₄ 2H ₂ O: 0.967g, NaCl: 0.90g, Casein: 0.25%-0.75%, Trehalose: 3%-5%, Proclin 300: 0.1%-0.5%, and Triton X-100: 0.025%- 0.075%. 5. a kind of thymidine kinase 1 magnetic particle chemiluminescent detection reagent according to claim 1, is characterized in that, described TK1 calibration product and TK1 quality control product are all made of phosphate buffer saline of 0.01mol/L, 0.25 %-0.75% casein and recombinant human TK1 protein formulated. 6. A kind of thymidine kinase 1 magnetic particle chemiluminescence detection reagent according to claim 1, characterized in that, the dilution liquid of the calibration product and the dilution liquid of the sample are formulated into: KCl: 0.30g, NaCl: 12.00g, KH ₂ PO ₄ : 0.30g, Na ₂ HPO ₄ : 1.725g, PH: 7.2-7.6, casein: 0.25%-0.75%, deionized water: 1500ml. 7. A method for preparing a thymidine kinase 1 magnetic particle chemiluminescent detection kit, comprising the following steps: Step 1: Preparation of magnetic particle suspension coated with anti-TK1 antibody: Add Tris-HCl buffer salt, sodium chloride, surfactant, trehalose, bovine serum albumin, and preservatives in deionized water in sequence. Each time you add materials, you need to stir until they are completely dissolved, adjust the pH value to 6.0-8.5, and mix After uniformity, pass through a 0.22 μm filter membrane to obtain the magnetic microsphere dilution, and then add the magnetic microsphere-labeled TK1 antibody conjugate to obtain the magnetic particle suspension coated with anti-TK1 antibody; Step 2: Preparation of detection reagents for acridinium ester-labeled anti-TK1 detection antibodies: Add phosphate buffer salt, surfactant, casein, trehalose, and preservatives in deionized water in sequence. Each addition needs to be stirred until completely dissolved, and the pH value is adjusted to 6.0-8.5. After mixing evenly, pass through a 0.22 μm filter membrane Obtain the diluent of the luminescent substance, and then add the acridinium ester-labeled anti-TK1 detection antibody conjugate to obtain the detection reagent of the acridinium ester-labeled anti-TK1 detection antibody; Step 3: Preparation of TK1 calibrator and TK1 quality control Add phosphate buffer salt, casein, and preservatives in deionized water in sequence. Each time you add materials, you need to stir until they are completely dissolved. Adjust the pH value to 6.0-8.5. Add the recombinant human TK1 protein to obtain the corresponding calibrator series and quality control series; Step 4: Assembling the kit: the above prepared magnetic particle suspension coated with anti-TK1 antibody and the detection reagent of anti-TK1 detection antibody labeled with acridinium ester in volume ratio R1:R2=1:1, and the corresponding calibration Product and quality control product series, the volume ratio is 1:1, divided into bottles to form a kit.

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