CN117538532A - Kit for detecting lung cancer antigen concentration by magnetic particle chemiluminescence immunoassay and detection method - Google Patents
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- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
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- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
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Abstract
The application provides a kit for detecting lung cancer antigen concentration by a magnetic particle chemiluminescence immunoassay and a detection method, wherein the kit comprises monoclonal antibodies respectively coupled with magnetic particles, wherein the monoclonal antibodies comprise a CA125 monoclonal antibody, a CEA monoclonal antibody, a CA199 monoclonal antibody, an NSE monoclonal antibody, a CY211 monoclonal antibody and an SCC monoclonal antibody; the kit also included a CA125 monoclonal antibody labeled with FITC and alkaline phosphatase, a CEA monoclonal antibody labeled with Cy3 and alkaline phosphatase, a CA199 monoclonal antibody labeled with Cy5 and alkaline phosphatase, an NSE monoclonal antibody labeled with DyLight549 and alkaline phosphatase, a Cy211 monoclonal antibody labeled with rhodoamineb and alkaline phosphatase, a SCC monoclonal antibody labeled with GFP and alkaline phosphatase. The kit has the advantages of simplicity, convenience, high sensitivity and the like.
Description
Technical Field
The application relates to the technical field of biological medicine, in particular to a kit for detecting lung cancer antigen concentration by a magnetic particle chemiluminescence immunoassay method and a detection method.
Background
Lung cancer is a malignant tumor that originates from abnormal cells in lung tissue and can proliferate and spread rapidly. Lung cancer can be divided into two main categories, depending on the type and location of the tumor: non-small cell lung cancer (NSCLC) and Small Cell Lung Cancer (SCLC).
Non-small cell lung cancer (NSCLC): NSCLC is the most common type of lung cancer, accounting for about 85% of all lung cancer cases. It includes adenocarcinomas, squamous carcinomas and large cell carcinomas. NSCLC generally grows slower and symptoms such as cough, expectoration, chest pain, dyspnea, etc. appear earlier. NSCLC may spread to other organs (e.g., liver, bone, brain, etc.) and form metastases if not treated in time.
Small Cell Lung Cancer (SCLC): SCLC accounts for about 15% of all lung cancer cases, grows rapidly and is strongly invasive. SCLC tends to spread early on to other sites such as lymph nodes, liver, bones and brain. Symptoms include cough, dyspnea, chest pain, weight loss, etc. Because of the difficulty in early diagnosis of SCLC, most patients are already in an advanced stage when diagnosed. The malignant nature of lung cancer makes it have the following serious consequences. Therefore, it is particularly important for early diagnosis and treatment of lung cancer.
Currently, in the prior art, a plurality of methods for detecting lung cancer exist, but the methods have advantages and disadvantages. X-ray examination is simple and low in cost, but has low sensitivity to early lung cancer, and small tumor cannot be accurately detected.
CT scanning, while capable of providing more accurate images, is more sensitive to early lung cancer, has a higher radiation dose and higher cost, and can lead to misdiagnosis and over-treatment.
Chest MRI, while having no radiation, is more accurate for lung cancer detection in some cases, but is more costly and therefore not a routine method for lung cancer detection.
PET-CT scans, while capable of providing information about tumor metabolism and location, are more sensitive to early stage lung cancer, are costly, have higher radiation doses, and can lead to misdiagnosis and over-treatment problems.
Disclosure of Invention
In order to solve at least one technical problem, a detection method which is rapid, simple, low in cost, high in sensitivity and high in detection accuracy and can assist in lung cancer diagnosis is developed, and the application provides a kit and a detection method for detecting lung cancer antigen concentration by a magnetic particle chemiluminescence immunoassay.
In a first aspect, the present application provides a kit for detecting the concentration of an antigen of lung cancer by a magnetic particle chemiluminescence immunoassay, wherein the kit comprises a buffer solution, a lung cancer marker A, a lung cancer marker B and a chemiluminescent substrate;
the lung cancer marker A comprises A1, A2, A3, A4, A5 and A6;
a1 includes a CA125 monoclonal antibody conjugated to magnetic particles;
a2 includes CEA monoclonal antibodies conjugated to magnetic microparticles;
a3 includes CA199 monoclonal antibodies coupled to magnetic particles;
a4 includes NSE monoclonal antibodies coupled to magnetic particles;
a5 includes CY211 monoclonal antibodies coupled to magnetic particles;
a6 includes SCC monoclonal antibodies coupled to magnetic particles;
the lung cancer marker B comprises B1, B2, B3, B4, B5 and B6;
b1 comprises a CA125 monoclonal antibody labeled with FITC and alkaline phosphatase;
b2 comprises CEA monoclonal antibody labeled with Cy3 and alkaline phosphatase;
b3 includes a CA199 monoclonal antibody labeled with Cy5 and alkaline phosphatase;
b4 includes NSE monoclonal antibodies labeled by DyLight549 and alkaline phosphatase;
b5 comprises CY211 monoclonal antibody marked by rhodomine B and alkaline phosphatase;
b6 includes SCC monoclonal antibodies labeled with GFP and alkaline phosphatase;
the concentrations of A1, A2, A3, A4, A5 and A6 are 300-500 mug/mL;
the concentrations of B1, B2, B3, B4, B5 and B6 are 200-300 mug/mL;
the concentration of the buffer solution is 0.1-0.2 mol/L.
Through the technical scheme, the method and the device can provide auxiliary detection for early diagnosis of lung cancer, can detect tumor markers with very low concentration, and have high sensitivity, so that accurate results can be provided; the specific antibody is combined with the target tumor marker, so that the specific tumor antigen can be accurately identified and measured, the interference of other substances is avoided, and the accuracy and the reliability of a detection result are improved; the biological samples such as serum or plasma are used for detection, and the samples are relatively easy and noninvasive to collect, so that the patients are not excessively uncomfortable. Meanwhile, the samples are relatively stable, so that the samples are convenient to store and transport, and the reliability of detection results is ensured; in general, the method has the advantages of high sensitivity, high specificity, rapidness, easiness in operation, specimen reliability and the like, and has important clinical significance for early auxiliary diagnosis, disease monitoring and treatment effect evaluation of lung cancer.
Alternatively, the chemiluminescent substrate comprises AMPPD at a concentration of 10-20mg/mL.
Optionally, the kit further comprises one or more of a calibrator, a quality control, and a sample diluent.
Optionally, the storage temperature of the kit for detecting the lung cancer related antigen concentration by the magnetic particle chemiluminescence immunoassay is 2-8 ℃.
In a second aspect, the present application provides a method for detecting the concentration of an antigen of lung cancer by using a magnetic particle chemiluminescence immunoassay, wherein the method comprises the following steps:
s1, respectively dissolving A1, A2, A3, A4, A5 and A6 in Tris buffer solution with the concentration of 0.1-0.2 mol/L, adding a sample to be detected, mixing, incubating to form an antibody-antigen complex, and cleaning;
dissolving B1, B2, B3, B4, B5 and B6 in Tris buffer solution with the concentration of 0.1-0.2 mol/L respectively for standby;
s2, after washing, adding B1 obtained in the step S1 into a reaction tube filled with A1, adding B2 obtained in the step S1 into a reaction tube filled with A2, adding B3 obtained in the step S1 into a reaction tube filled with A3, adding B4 obtained in the step S1 into a reaction tube filled with A4, adding B5 obtained in the step S1 into a reaction tube filled with A5, and adding B6 obtained in the step S1 into a reaction tube filled with A6, and performing a reaction to form an antibody-antigen-antibody sandwich complex;
s3, after the reaction is finished, washing unreacted substances by using a washing liquid;
s4, after washing, adding a luminescent substrate into each reaction tube to trigger a chemiluminescent reaction;
s5, detecting the intensity of the luminous signal.
Through adopting above-mentioned technical scheme, this application has highly automated characteristics, can handle a plurality of samples simultaneously, improves detection efficiency. Meanwhile, the detection time of the technology is shorter, and the result can be obtained in a shorter time. In addition, in the detection of lung cancer, the fluorescence signal and the enzyme reaction signal can be obtained simultaneously by using a detection method of multiple markers of an antibody marked by fluorescent dye and enzyme, so that multiple detection can be performed in cells or tissues. The strategy of multiple labeling can provide more information, and increase the flexibility and accuracy of experiments, so that the distribution and expression of target molecules can be more comprehensively known.
Optionally, the sample to be tested in step S2 includes one of human serum and plasma.
Optionally, the mixing incubation parameters in step S2 are as follows:
the incubation temperature is 20-25 ℃; incubation time is 30-60 min; the pH value during incubation is 7.2-7.4.
Optionally, the washing solution in step S1 and step S3 includes a phosphate buffer solution; the number of washes was 2.
In a third aspect, the application provides an application of the kit for detecting lung cancer antigen concentration by using the magnetic particle chemiluminescence immunoassay in the field of lung cancer detection.
In summary, the present invention includes at least one of the following beneficial technical effects:
1. the kit for detecting the lung cancer antigen concentration by using the magnetic particle chemiluminescence immunoassay has the advantages of high sensitivity, high specificity, rapidness, easiness in operation, specimen reliability and the like, and has important clinical significance for early auxiliary diagnosis, disease monitoring and treatment effect evaluation of lung cancer.
2. The method has the characteristic of high automation, can process a plurality of samples simultaneously, and improves detection efficiency. Meanwhile, the detection time of the technology is shorter, and the result can be obtained in a shorter time. In addition, in the detection of lung cancer, the fluorescence signal and the enzyme reaction signal can be obtained simultaneously by using a detection method of multiple markers of an antibody marked by fluorescent dye and enzyme, so that multiple detection can be performed in cells or tissues. The strategy of multiple labeling can provide more information, and increase the flexibility and accuracy of experiments, so that the distribution and expression of target molecules can be more comprehensively known.
Detailed Description
The present application is described in further detail below with reference to examples.
The following are examples of the preparation of the present application
Preparation example 1
The preparation method prepares a magnetic particle coupled lung cancer marker A, wherein the magnetic particle coupled lung cancer marker A comprises A1, A2, A3, A4, A5 and A6;
wherein A1 is a CA125 monoclonal antibody coupled with magnetic particles; a2 is CEA monoclonal antibody coupled with magnetic particles; a3 is a CA199 monoclonal antibody conjugated to magnetic particles; a4 is NSE monoclonal antibody coupled to magnetic particles; a5 is a CY211 monoclonal antibody conjugated to magnetic particles; a6 is an SCC monoclonal antibody coupled to magnetic particles;
the preparation method of the magnetic particle coupled monoclonal antibody comprises the following steps:
1. preparing magnetic particles: selecting magnetic particles with carboxyl groups on the surfaces;
(2) Washing the magnetic particles: washing the magnetic particles to remove potential contaminants and unbound materials to ensure purity and specificity of the subsequent binding step;
(3) Activating magnetic particles: activating active groups on the surface of the magnetic particles using N-hydroxysuccinimide to covalently bind the monoclonal antibodies thereto;
1. covalently binding monoclonal antibodies: respectively carrying out covalent binding on the CA125 monoclonal antibody, the CEA monoclonal antibody, the CA199 monoclonal antibody, the NSE monoclonal antibody, the CY211 monoclonal antibody and the SCC monoclonal antibody and the activated magnetic particles; obtaining A1, A2, A3, A4, A5 and A6; (the principle is that the carboxyl groups on the surface of the magnetic particles are subjected to covalent bonding with amino groups in the antibody;
2. blocking agent treatment: to reduce non-specific binding, the magnetic microparticles are treated with bovine serum albumin, blocking unbound active sites;
3. washing and purifying: washing the magnetic particles by using phosphate buffer solution to remove non-specifically bound substances so as to improve the specificity and purity of the binding;
(7) Identification and quality control: after washing, the enzyme-linked immunosorbent assay is adopted to identify and control the quality of A1, A2, A3, A4, A5 and A6, so as to ensure the stable combination, the specificity and the activity.
Preparation example 2
The lung cancer marker B is prepared by the preparation example, and comprises B1, B2, B3, B4, B5 and B6.
B1 is a CA125 monoclonal antibody marked by FITC and alkaline phosphatase;
b2 is CEA monoclonal antibody labeled by Cy3 and alkaline phosphatase;
b3 is a CA199 monoclonal antibody marked by Cy5 and alkaline phosphatase;
b4 is NSE monoclonal antibody labeled by DyLight549 and alkaline phosphatase;
b5 is CY211 monoclonal antibody marked by rhodomine B and alkaline phosphatase;
b6 is SCC monoclonal antibody marked by GFP and alkaline phosphatase;
the preparation method of B1, B2, B3, B4, B5 and B6 comprises the following steps:
1. monoclonal antibody cell lines: selecting a CA125 monoclonal antibody cell strain, a CEA monoclonal antibody cell strain, a CA199 monoclonal antibody cell strain, an NSE monoclonal antibody cell strain, a CY211 monoclonal antibody cell strain and an SCC monoclonal antibody cell strain for later use;
2. monoclonal antibody culture: adding each monoclonal antibody cell strain in the step (1) into DMEM added with FBS, culturing and amplifying in a constant temperature incubator at 37 ℃ with the pH value of the DMEM controlled between 7.2 and 7.4;
(2) Antibody purification: purifying the amplified antibody from the cell culture supernatant by protein L chromatography to remove other proteins and impurities;
(3) Marking: adding a fluorescent dye labeling group into the purified antibody to obtain a fluorescent dye labeled antibody, which is specifically as follows:
adding FITC solution into the purified CA125 monoclonal antibody for conjugation reaction for 30min to obtain a CA125 monoclonal antibody marked by FITC;
adding the Cy3 solution into the purified CEA monoclonal antibody for conjugation reaction for 30min to obtain a CEA monoclonal antibody marked by Cy 3;
adding the Cy5 solution into the CA199 monoclonal antibody which is subjected to purification, and carrying out conjugation reaction for 30min to obtain the CA199 monoclonal antibody marked by Cy 5;
adding Dylight549 solution into the NSE monoclonal antibody after purification to perform conjugation reaction for 30min, so as to obtain NSE monoclonal antibody marked by Dylight 549;
adding the Rhodamine B solution into the CY211 monoclonal antibody after purification for conjugation reaction for 30min to obtain a CY211 monoclonal antibody marked by Rhodamine B;
adding GFP solution into the purified SCC monoclonal antibody for conjugation reaction for 30min to obtain the SCC monoclonal antibody marked by GFP;
reaction termination: after the completion of the reaction, the reaction with the antibody was stopped by adding an appropriate amount of aminopolyglycerol.
Alkaline phosphatase labeling: alkaline phosphatase labeling is carried out on the solution of the fluorescent dye labeled antibody respectively, and the method is concretely as follows:
firstly, mixing a fluorescent dye-labeled antibody with a buffer solution (PBS) buffer solution to obtain a fluorescent dye-labeled antibody solution, and then adding alkaline phosphatase solution into each fluorescent dye-labeled antibody solution to react for 30min to obtain antibodies labeled by fluorescent dye and alkaline phosphatase, wherein the antibodies are respectively B1, B2, B3, B4, B5 and B6;
reaction termination: appropriate bovine serum albumin is added to stop the reaction of alkaline phosphatase with the antibody.
Split charging and storage: packaging labeled B1, B2, B3, B4, B5, and B6 into proper containers, and preserving by avoiding illumination and refrigeration to maintain activity and stability.
The following are specific examples of the present application
Examples 1 to 3
Example 1
The embodiment provides a kit for detecting the concentration of lung cancer antigens by a magnetic particle chemiluminescence immunoassay, which comprises a buffer solution, a washing solution, a lung cancer marker A, a lung cancer marker B and a chemiluminescent substrate;
wherein the lung cancer marker A comprises A1, A2, A3, A4, A5 and A6;
wherein the lung cancer marker B comprises B1, B2, B3, B4, B5 and B6;
the preparation method of the lung cancer marker A is shown in preparation example 1; the preparation method of the lung cancer marker B is shown in preparation example 2;
a1 is a magnetic particle coupled CA125 monoclonal antibody;
a2 is CEA monoclonal antibody coupled with magnetic particles;
a3 is a CA199 monoclonal antibody conjugated to magnetic particles;
a4 is NSE monoclonal antibody coupled to magnetic particles;
a5 is a CY211 monoclonal antibody conjugated to magnetic particles;
a6 is an SCC monoclonal antibody coupled to magnetic particles;
the lung cancer marker B comprises B1, B2, B3, B4, B5 and B6;
b1 is a CA125 monoclonal antibody marked by FITC and alkaline phosphatase;
b2 is CEA monoclonal antibody labeled by Cy3 and alkaline phosphatase;
b3 is a CA199 monoclonal antibody marked by Cy5 and alkaline phosphatase;
b4 is NSE monoclonal antibody labeled by DyLight549 and alkaline phosphatase;
b5 is CY211 monoclonal antibody marked by rhodomine B and alkaline phosphatase;
b6 is SCC monoclonal antibody marked by GFP and alkaline phosphatase;
the concentrations of A1, A2, A3, A4, A5 and A6 are 300 mug/mL;
the concentrations of B1, B2, B3, B4, B5 and B6 are all 0.4 mug/mL;
the buffer concentration was 0.1mol/L.
The concentrations and compositions of the components of this example are shown in Table 1;
examples 2 to 3
Examples 2 to 3 differ from example 1 in the concentration of each component, see Table 1 for the differences.
TABLE 1 concentration of the components of examples 1-3
Accuracy test, experiments 1-3
Experiment 1
The present experiment uses a kit for detecting lung cancer antigen concentration by using a magnetic particle chemiluminescence immunoassay obtained in example 1 to detect lung cancer antigen concentration of 5 human blood samples (respectively, 1, 2, 3, 4 and 5 samples) with known detection results, and the detection method comprises the following steps:
s1, respectively dissolving A1, A2, A3, A4, A5 and A6 in Tris buffer solution with the concentration of 0.1mol/L, adding 5000U/mL of human blood sample, mixing and incubating (the incubation temperature is 37 ℃, the incubation time is 30min, and the pH value during incubation is 7.2-7.4) to form an antibody-antigen complex, and washing twice with phosphate buffer solution;
dissolving B1, B2, B3, B4, B5 and B6 in Tris buffer solution with the concentration of 0.1-0.2 mol/L respectively for standby;
s2, after washing, adding B1 obtained in the step S1 into a reaction tube filled with A1, adding B2 obtained in the step S1 into a reaction tube filled with A2, adding B3 obtained in the step S1 into a reaction tube filled with A3, adding B4 obtained in the step S1 into a reaction tube filled with A4, adding B5 obtained in the step S1 into a reaction tube filled with A5, and adding B6 obtained in the step S1 into a reaction tube filled with A6, and performing a reaction to form an antibody-antigen-antibody sandwich complex;
s3, after the reaction is finished, washing unreacted substances by using a washing liquid;
s4, after washing, adding a chemiluminescent substrate (AMPPD) into each reaction tube to trigger a chemiluminescent reaction;
s5, detecting the intensity of the luminous signal, and comparing the luminous signal with the quality control product.
The preparation method of the quality control product comprises the following steps: and respectively dissolving the CA125 antigen, the CEA antigen, the CA199 antigen, the NSE antigen, the CY211 antigen and the SCC antigen in 0.1mol/mLTris buffer solution to obtain quality control products of the antigens.
The preparation method of the calibrator comprises the following steps: the CA125 antigen, CEA antigen, CA199 antigen, NSE antigen, CY211 antigen and SCC antigen are respectively dissolved in 0.1mol/mLTris buffer solution to obtain the calibrator of each antigen.
The target value range of the calibrator (50.00+/-7.50) U/mL and (1000.00+/-150.00) U/mL.
The target value range of the quality control product (30.00+/-4.50) U/mL and the target value range of the quality control product (500.00+/-75.00) U/mL.
The known test results of the 5 human blood samples of the known test results are shown in table 2, wherein (v) indicates that the blood sample contains the corresponding antigen and (- -) indicates that the corresponding antigen is not contained.
TABLE 2 known detection results of human blood samples for known detection results see Table
The test results of experiment 1 are shown in table 3;
TABLE 3 detection results of experiment 1
Experiment 2
The present experiment was different from experiment 1 in that it was performed using a kit for detecting the concentration of lung cancer antigen by the magnetic particle chemiluminescence immunoassay obtained in example 2. The experimental detection results of experiment 2 are shown in table 4.
TABLE 4 detection results of experiment 2
Experiment 3
The present experiment was different from experiment 1 in that it was performed using a kit for detecting the concentration of lung cancer antigen by the magnetic particle chemiluminescence immunoassay obtained in example 3. The experimental detection results of experiment 3 are shown in table 5.
TABLE 5 detection results of experiment 3
Referring to tables 2 to 5, the detection results of experiments 1 to 3 are consistent with the known detection results of blood samples, so that the kit for detecting the lung cancer antigen concentration by the magnetic particle chemiluminescence immunoassay provided by the application has the advantage of high detection accuracy. Meanwhile, the kit for detecting the lung cancer antigen concentration by the magnetic particle chemiluminescence immunoassay has the advantages that in the first aspect, the kit can detect various lung cancer antigens simultaneously, comprehensive tumor marker information is provided, and the kit is favorable for more comprehensively evaluating the tumor state.
In the second aspect, the specificity of the antigen can be further verified by detecting the wavelength of the antigen, so that the accuracy of a detection result is ensured.
In the third aspect, the magnetic particle chemiluminescence immunoassay has the characteristics of high speed and high efficiency, and can complete the detection process in a relatively short time, thereby improving the detection efficiency. In conclusion, the kit for detecting the lung cancer antigen concentration by the magnetic particle chemiluminescence immunoassay has the advantages of high accuracy, multiple antigen detection, wavelength detection, rapidness, simplicity and convenience and the like, and is beneficial to providing accurate and comprehensive lung cancer antigen concentration information.
Sensitivity test, experiment 4
The experiment was different from experiment 1 in that the concentration of sample No. 1 was diluted to 4000.00U/mL, 100.00U/mL, 60.00U/mL, 10.00U/mL, 1.00U/mL, and 0.500U/mL, respectively, for detection.
The results of this sensitivity test are shown in Table 6.
TABLE 6-experiment 4 sensitivity test results see Table
Analysis of results:
referring to the sensitivity detection experimental results provided in table 6, the kit for detecting the lung cancer antigen concentration by the magnetic particle chemiluminescence immunoassay provided by the application can accurately detect samples with the concentration not less than 1.00U/mL when detecting samples to be detected.
The foregoing are all preferred embodiments of the present application, and are not intended to limit the scope of the present application in any way, therefore: all equivalent changes in structure, shape and principle of this application should be covered in the protection scope of this application.
Claims (9)
1. The kit for detecting the lung cancer antigen concentration by using a magnetic particle chemiluminescence immunoassay is characterized by comprising a buffer solution, a lung cancer marker A, a lung cancer marker B and a chemiluminescent substrate;
the lung cancer marker A comprises A1, A2, A3, A4, A5 and A6;
a1 includes a CA125 monoclonal antibody conjugated to magnetic particles;
a2 includes CEA monoclonal antibodies conjugated to magnetic microparticles;
a3 includes CA199 monoclonal antibodies coupled to magnetic particles;
a4 includes NSE monoclonal antibodies coupled to magnetic particles;
a5 includes CY211 monoclonal antibodies coupled to magnetic particles;
a6 includes SCC monoclonal antibodies coupled to magnetic particles;
the lung cancer marker B comprises B1, B2, B3, B4, B5 and B6;
b1 comprises a CA125 monoclonal antibody labeled with FITC and alkaline phosphatase;
b2 comprises CEA monoclonal antibody labeled with Cy3 and alkaline phosphatase;
b3 includes a CA199 monoclonal antibody labeled with Cy5 and alkaline phosphatase;
b4 includes NSE monoclonal antibodies labeled by DyLight549 and alkaline phosphatase;
b5 comprises CY211 monoclonal antibody marked by rhodomine B and alkaline phosphatase;
b6 includes SCC monoclonal antibodies labeled with GFP and alkaline phosphatase;
the concentrations of A1, A2, A3, A4, A5 and A6 are 300-500 mug/mL;
the concentrations of B1, B2, B3, B4, B5 and B6 are 200-300 mug/mL;
the concentration of the buffer solution is 0.1-0.2 mol/L.
2. The kit for detecting the concentration of the lung cancer antigen by the magnetic particle chemiluminescence immunoassay according to claim 1, wherein the chemiluminescent substrate comprises AMPPD with the concentration of 10-20mg/mL.
3. The kit for detecting the concentration of the lung cancer antigen by the magnetic particle chemiluminescence immunoassay according to claim 1, wherein the kit further comprises one or more of a calibrator, a quality control material and a sample diluent.
4. The kit for detecting the lung cancer antigen concentration by using the magnetic particle chemiluminescence immunoassay according to claim 1, wherein the storage temperature of the kit for detecting the lung cancer-related antigen concentration by using the magnetic particle chemiluminescence immunoassay is 2-8 ℃.
5. A method for detecting the concentration of an antigen of lung cancer by a magnetic particle chemiluminescence immunoassay according to claim 1, wherein the method comprises the following steps:
s1, respectively dissolving A1, A2, A3, A4, A5 and A6 in Tris buffer solution with the concentration of 0.1-0.2 mol/L, adding a sample to be tested, mixing, incubating to form an antibody-antigen complex, and cleaning;
dissolving B1, B2, B3, B4, B5 and B6 in Tris buffer solution with the concentration of 0.1-0.2 mol/L respectively for standby;
s2, after washing, adding B1 obtained in the step S1 into a reaction tube filled with A1, adding B2 obtained in the step S1 into a reaction tube filled with A2, adding B3 obtained in the step S1 into a reaction tube filled with A3, adding B4 obtained in the step S1 into a reaction tube filled with A4, adding B5 obtained in the step S1 into a reaction tube filled with A5, and adding B6 obtained in the step S1 into a reaction tube filled with A6, and performing a reaction to form an antibody-antigen-antibody sandwich complex;
s3, after the reaction is finished, washing unreacted substances by using a washing liquid;
s4, after washing, adding a luminescent substrate into each reaction tube to trigger a chemiluminescent reaction;
s5, detecting the intensity of the luminous signal.
6. The method according to claim 5, wherein the sample to be tested in step S1 comprises one of human serum and plasma.
7. The method for detecting lung cancer antigen concentration by magnetic particle chemiluminescence immunoassay according to claim 5, wherein the mixed incubation parameters in step S1 are as follows:
incubation temperature was 37 ℃; incubation time is 30-60 min; the pH value during incubation is 7.2-7.4.
8. The method for detecting lung cancer antigen concentration by magnetic particle chemiluminescence immunoassay according to claim 5, wherein the washing solution in step S1 and step S3 comprises phosphate buffer solution; the number of washes was 2.
9. Use of the kit for detecting lung cancer antigen concentration by using the magnetic particle chemiluminescence immunoassay of claim 1 in the field of lung cancer detection.
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