Magnetic particle chemiluminescence kit for NSE correction detection of hemolysis sample and NSE detection method

Abstract

The invention provides a magnetic particle chemiluminescence kit for NSE correction detection of a hemolysis sample and an NSE detection method, and belongs to the technical field of biochemical detection. The kit comprises a magnetic particle coated by an NSE monoclonal antibody, a magnetic particle coated by a hemoglobin monoclonal antibody, an NSE monoclonal antibody marked by acridinium ester, a hemoglobin monoclonal antibody marked by acridinium ester, an NSE series calibrator and a hemoglobin series calibrator. The kit can accurately detect the NSE content in a hemolysis sample, and the accuracy of a detection result is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of biochemical detection, and particularly relates to a magnetic particle chemiluminescence kit for NSE correction detection of hemolytic samples and an NSE detection method. Background Art

[0002] Neuron-specific enolase (NSE) is a cell-specific isoenzyme of glycolytic enzymes, mainly present in neurons and neuroendocrine cells. As an important tumor marker, NSE is of great significance in the diagnosis and monitoring of diseases such as neuroendocrine tumors and small cell lung cancer. In practical applications, the determination of NSE is usually combined with other clinical information and diagnostic tools to improve the accuracy of diagnosis and the reliability of prognostic assessment.

[0003] However, in clinical tests, hemolyzed samples can interfere with a variety of test items, especially the detection of biochemical indicators such as NSE, due to the destruction of red blood cells and the release of hemoglobin. In addition, because red blood cells contain a high concentration of NSE, hemolysis can cause these NSE to be released into the serum, causing the NSE test value to increase abnormally, resulting in a false positive result. In clinical practice, if you encounter an abnormally high level of NSE, you need to consider the possibility of hemolysis. If hemolysis is suspected, you should contact the clinic to determine whether you need to re-draw blood for reexamination to ensure the accuracy of the test results. Therefore, in order to avoid this interference, the collected blood samples should be centrifuged and tested as soon as possible to reduce the impact of the release of red blood cell contents on the test results.

[0004] In summary, providing a method or reagent that is useful for correcting the concentration of specific enolase in hemolyzed samples is a problem that we urgently need to solve. Summary of the invention

[0005] In order to solve the problem of abnormally increased concentration of specific enolase in hemolyzed samples, the present invention provides a magnetic particle chemiluminescence kit for NSE correction detection of hemolyzed samples, which can accurately detect the NSE content in hemolyzed samples and improve the accuracy of the detection results.

[0006] The present invention also provides a NSE detection method based on back calculation of hemoglobin content.

[0007] The present invention is achieved through the following technical solutions:

[0008] The invention provides a magnetic particle chemiluminescence kit for NSE calibration detection of hemolyzed samples. The kit comprises magnetic particles coated with NSE monoclonal antibodies, magnetic particles coated with hemoglobin monoclonal antibodies, acridinium ester-labeled NSE monoclonal antibodies, acridinium ester-labeled hemoglobin monoclonal antibodies, NSE series calibrators and hemoglobin series calibrators.

[0009] Furthermore, in the NSE series calibrators, the concentration range of NSE is 1 to 300 ng / mL;

[0010] In the hemoglobin series calibrators, the hemoglobin concentration range is 0 to 200 g / L.

[0011] Furthermore, the NSE monoclonal antibody-coated magnetic particles are suspended in a blocking storage solution to prepare a NSE monoclonal antibody-coated magnetic particle solution with a concentration of 0.1 to 0.4 μg / mL;

[0012] The magnetic particles coated with the hemoglobin monoclonal antibody are suspended in the closed storage solution to prepare a hemoglobin monoclonal antibody coated magnetic particle solution with a concentration of 0.1 to 0.4 μg / mL;

[0013] The acridinium ester-labeled NSE monoclonal antibody is dissolved in a labeling diluent to prepare an acridinium ester-labeled NSE monoclonal antibody solution with a concentration of 0.05 to 0.5 μg / mL;

[0014] The acridinium ester-labeled hemoglobin monoclonal antibody is dissolved in the labeling diluent to prepare an acridinium ester-labeled hemoglobin monoclonal antibody solution with a concentration of 0.05 to 0.5 μg / mL;

[0015] The closed preservation solution contains the following ingredients:

[0016] Tris-HCl: 0.02-0.1M; Bovine serum albumin (BSA): 0.5-3wt%; ProCl in300 (liquid biological preservative): 0.05-0.3wt%; Triton X100: 0.05-0.3wt%; Sodium chloride: 0.5-0.9wt%;

[0017] The pH value of the sealing storage solution is 7 to 8;

[0018] The labeling diluent contains the following ingredients:

[0019] Tris-HCl: 0.02~0.1M; Bovine serum albumin: 0.5~3wt%; ProCl in300: 0.05~0.3wt%; Triton X100: 0.05~0.3wt%; Magnesium chloride: 0.1~0.5wt%.

[0020] Furthermore, the NSE monoclonal antibody-coated magnetic microparticles are prepared by covalently binding the NSE monoclonal antibody to the surface of the carboxyl-modified magnetic microparticles;

[0021] The hemoglobin monoclonal antibody-coated magnetic particles are prepared by covalently binding the hemoglobin monoclonal antibody to the surface of the carboxyl-modified magnetic particles;

[0022] The acridinium ester-labeled NSE monoclonal antibody and the acridinium ester-labeled hemoglobin monoclonal antibody are both prepared by dialysis;

[0023] The NSE series calibrants are prepared by diluting NSE with a bovine serum albumin solution and a MgCl solution;

[0024] The hemoglobin series calibrants are prepared by diluting hemoglobin with a bovine serum albumin solution and a MgCl solution.

[0025] Further, the NSE monoclonal antibody includes NSE mouse monoclonal antibody;

[0026] The hemoglobin monoclonal antibody includes a human hemoglobin monoclonal antibody (hHb (H5A3) Mouse mAb).

[0027] Based on the same inventive concept, the present invention provides an application of a magnetic particle chemiluminescence kit for NSE correction detection of hemolyzed samples in the detection of NSE content in hemolyzed samples for non-disease diagnosis purposes.

[0028] Based on the same inventive concept, the present invention provides a NSE detection method based on hemoglobin content back calculation, the detection method comprising:

[0029] The NSE concentration of the hemolyzed sample was tested using a magnetic particle chemiluminescence kit to obtain preliminary NSE concentration test results;

[0030] Using the magnetic particle chemiluminescence kit to detect the hemoglobin concentration of the hemolyzed sample to obtain a hemoglobin concentration detection result;

[0031] According to the hemoglobin concentration detection result and the interference model, the preliminary NSE concentration detection result is back-calculated and corrected to obtain a corrected NSE concentration detection value;

[0032] Wherein, the magnetic particle chemiluminescence kit includes the above-mentioned magnetic particle chemiluminescence kit for NSE correction detection of hemolytic samples.

[0033] Furthermore, the NSE concentration of the hemolyzed sample is detected by using a magnetic particle chemiluminescence kit to obtain a preliminary detection result of the NSE concentration, specifically including:

[0034] The NSE concentration of the hemolyzed sample was detected using magnetic particles coated with NSE monoclonal antibody and acridinium ester-labeled NSE monoclonal antibody in the magnetic particle chemiluminescence kit to obtain preliminary detection results of NSE concentration.

[0035] Furthermore, the method of detecting the hemoglobin concentration of the hemolyzed sample using the magnetic particle chemiluminescence kit to obtain the hemoglobin concentration detection result specifically includes:

[0036] The hemoglobin concentration of the hemolyzed sample is detected using the magnetic particles coated with the hemoglobin monoclonal antibody and the hemoglobin monoclonal antibody labeled with acridinium ester in the magnetic particle chemiluminescence kit to obtain the hemoglobin concentration detection result.

[0037] Further, the preliminary NSE concentration detection result is back-calculated and corrected according to the hemoglobin concentration detection result and the interference model to obtain a corrected NSE concentration detection value, specifically including:

[0038] The interference model formula is as follows:

[0039] y=-0.8053x 2 +36.589x-4.3696, where x is the hemoglobin concentration in the hemolyzed serum, and y is the total concentration of NSE released by hemoglobin and erythrocytes in the hemolyzed serum;

[0040] Taking the hemoglobin concentration test result as x, substituting it into the interference model formula to obtain the y value;

[0041] The result obtained by subtracting the y value from the preliminary NSE concentration test result is the corrected NSE concentration test value.

[0042] One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages:

[0043] 1. The present invention discloses a magnetic microparticle chemiluminescence kit for NSE correction detection of hemolyzed samples. The magnetic microparticles coated with NSE monoclonal antibodies, acridinium ester-labeled NSE monoclonal antibodies and a closed preservation solution in the kit can detect the total NSE concentration in the hemolyzed samples. The magnetic microparticles coated with hemoglobin monoclonal antibodies, acridinium ester-labeled hemoglobin monoclonal antibodies and a closed preservation solution can detect the hemoglobin concentration in the hemolyzed samples. According to the hemoglobin concentration detection result and the interference model, the NSE concentration detection value after the total NSE concentration correction can be obtained. The kit can accurately detect the NSE content in the hemolyzed samples and improve the accuracy of the detection results.

[0044] 2. The present invention discloses an NSE detection method based on back-calculation of hemoglobin content. The method obtains the correlation between the hemoglobin concentration in hemolyzed serum and the total concentration of NSE released by hemoglobin and red blood cells by constructing an interference model formula, and then detects the preliminary detection result of NSE concentration and the hemoglobin concentration of the hemolyzed sample by the magnetic particle chemiluminescence kit of the present invention. According to the hemoglobin concentration detection result and the interference model formula, the preliminary detection result of NSE concentration is back-calculated and corrected to obtain a corrected NSE concentration detection value. The method can accurately evaluate the interference of hemoglobin in the hemolyzed sample on NSE detection, and improves the accuracy of NSE detection of the hemolyzed sample through back-calculation and correction of the hemoglobin content. The method is suitable for an automated detection platform and reduces operational complexity and human errors. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0046] Figure 1 This is the interference curve of pure hemoglobin on NSE concentration detection.

[0047] Figure 2 This is the relationship curve between hemoglobin and NSE concentration released by red blood cells.

[0048] Figure 3 This is the relationship curve between hemoglobin and the total concentration of interfering NSE. DETAILED DESCRIPTION

[0049] The present invention will be described in detail below in conjunction with specific implementations and examples, and the advantages and various effects of the present invention will be more clearly presented. It should be understood by those skilled in the art that these specific implementations and examples are used to illustrate the present invention, rather than to limit the present invention.

[0050] Throughout the specification, unless otherwise specifically stated, the terms used herein should be understood as meanings commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which the present invention belongs. In the event of a conflict, the present specification takes precedence.

[0051] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0052] The following will describe in detail a magnetic microparticle chemiluminescence kit and an NSE detection method for NSE correction detection of hemolytic samples in the present application in combination with examples and experimental data.

[0053] Example 1

[0054] This embodiment provides a method for preparing a magnetic microparticle chemiluminescence kit for NSE correction detection of hemolytic samples.

[0055] 1. Preparation of magnetic particles coated with NSE monoclonal antibody or hemoglobin monoclonal antibody.

[0056] (1) 100uL of carboxyl-modified magnetic microparticle stock solution (purchased from Beijing Boerma Biotechnology Co., Ltd. (MS160 / Carboxy l)) was added with 0.1M pH5.0 MES solution (4-morpholineethanesulfonic acid) to wash twice, and the supernatant was removed by applying a magnetic field; 300uL of 2mg / mL NHS (N-hydroxysuccinimide) and EDC (1-ethyl-(3-dimethylaminopropyl) carbodiimide) solution were taken respectively, mixed with the magnetic microparticles, and then oscillated for 2h, and a magnetic field was applied to remove the supernatant; 300uL of 0.1M pH5.0 MES solution was mixed with the magnetic microparticles, and then oscillated for 5min, and a magnetic field was applied to remove the supernatant, and the mixture was repeated twice.

[0057] (2) Add 300uL of 0.1M pH5.0 MES solution to the magnetic particles, mix well, add 0.3mg of NSE monoclonal antibody or hemoglobin monoclonal antibody (NSE mouse monoclonal antibody / human hemoglobin monoclonal antibody), mix well, shake for 5h, apply magnetic field to remove supernatant. Then add 300uL of 0.05M pH7.2 PB solution (phosphate buffer solution), shake for 5min, apply magnetic field to remove supernatant, and repeat twice.

[0058] (3) Add 300uL of stop solution prepared by mixing 0.05M pH7.2 PB solution and ethanolamine solution in a volume ratio of 5:1, mix well and shake for 2h, remove the supernatant by applying a magnetic field. Add 300uL of blocking solution (0.05M pH7.5 TB (Tris-HCl buffer) solution containing 2% bovine serum albumin (BSA), 0.9% sodium chloride (NaCl), 0.2% Triton X100 and 0.2% ProCl in 300 preservatives by weight), shake for 5min, remove the supernatant by applying a magnetic field, and repeat twice.

[0059] (4) Add 300uL of 0.05M pH7.5 TB solution containing 2% bovine serum albumin (BSA), 0.9% sodium chloride (NaCl), 0.2% Triton X100 and 0.2% ProCl in300 preservatives by mass ratio, shake for 12h, apply magnetic field to remove supernatant, and obtain magnetic particles coated with NSE monoclonal antibody or magnetic particles coated with hemoglobin monoclonal antibody. Use blocking preservation solution to prepare magnetic particles coated with NSE monoclonal antibody or magnetic particles coated with hemoglobin monoclonal antibody into a 0.2μg / mL magnetic particle solution. The prepared magnetic particle solution is stored at 2-8°C, should not be frozen, and should be fully mixed when used.

[0060] 2. Preparation of acridinium ester-labeled NSE monoclonal antibody or acridinium ester-labeled hemoglobin monoclonal antibody:

[0061] (1) Pre-experimental treatment: Take 0.1 mg of NSE monoclonal antibody or hemoglobin monoclonal antibody and dilute the antibody concentration to 0.5 mg / mL with 0.05 M PB solution (pH 7.2).

[0062] (2) Mix well at a molar ratio of 20:1 (acridinium ester:antibody) and allow to react at room temperature for 1 h.

[0063] (3) After the coupling reaction, the acridinium ester labeled product was transferred into a dialysis bag and immersed in a 0.05 M PB solution (pH 7.2) for more than 5 min. The product was dialyzed in the PB solution for 24 h. The purified liquid volume was measured and an equal volume of glycerol was added and mixed using a vortex machine.

[0064] (4) The prepared acridinium ester marker was stored at -20°C and returned to room temperature before use and mixed thoroughly.

[0065] (5) The prepared acridinium ester-labeled NSE monoclonal antibody is dissolved in a labeling diluent to prepare an acridinium ester-labeled NSE monoclonal antibody solution with a concentration of 0.2 μg / mL; the acridinium ester-labeled hemoglobin monoclonal antibody is dissolved in the labeling diluent to prepare an acridinium ester-labeled hemoglobin monoclonal antibody solution with a concentration of 0.2 μg / mL.

[0066] The labeling diluent contains the following ingredients:

[0067] Tris-HCl: 0.05M pH 7.5; bovine serum albumin: 2wt%; ProCl in 300: 0.2wt%; Triton X100: 0.2wt%; magnesium chloride: 0.3wt%.

[0068] 3. Preparation of blocking solution:

[0069] BSA, Triton X100, sodium chloride and ProClin300 were added to 0.05M Tris-HCl buffer (pH=7.5), wherein the mass concentration of each component was: 2% bovine serum albumin BSA, 0.2% ProCl in300, 0.2% Triton X100, and 0.9% sodium chloride.

[0070] 4. Preparation of NSE series calibrators and hemoglobin series calibrators.

[0071] The pure NSE was diluted into freeze-dried calibrators with concentrations of 0, 10, 50, 100, 200, and 300 ng / mL using 0.05 M pH 7.2 solution containing 2% bovine serum albumin (BSA) and 0.2% MgCl; the pure hemoglobin was diluted into freeze-dried calibrators with concentrations of 0, 30, 60, 100, 150, and 200 g / L using 0.05 M pH 7.2 PB solution containing 2% bovine serum albumin (BSA).

[0072] Example 2

[0073] A method for detecting NSE based on back calculation of hemoglobin content.

[0074] (1) Collect blood samples from patients and separate serum.

[0075] (2) The NSE concentration was preliminarily detected using a fully automatic chemiluminescence immunoassay instrument and an NSE magnetic microparticle chemiluminescence kit. The sample, magnetic microparticles coated with NSE monoclonal antibodies, and acridinium ester-labeled NSE monoclonal antibodies were incubated together. After the NSE in the sample was incubated with the monoclonal antibodies coated on the magnetic microparticles and the monoclonal antibodies labeled with acridinium ester, an immune complex was formed. Unbound substances were removed by magnetic separation and washing. Finally, a substrate solution was added to the reaction complex, and the relative light intensity (RLU) was measured. The concentration of NSE in the sample is proportional to the relative light intensity (RLU) measured by the optical system of the fully automatic chemiluminescence immunoassay instrument. The instrument automatically fits and calculates the concentration of NSE.

[0076] (3) The hemoglobin concentration in serum is detected by a fully automatic chemiluminescence immunoassay and a hemoglobin magnetic particle chemiluminescence kit. The sample, magnetic particles coated with hemoglobin monoclonal antibodies, and acridinium ester-labeled hemoglobin monoclonal antibodies are incubated together. After the hemoglobin in the sample is incubated with the monoclonal antibodies coated on the magnetic particles and the monoclonal antibodies labeled with acridinium ester, an immune complex is formed. Unbound substances are removed by magnetic separation and washing. Finally, a substrate solution is added to the reaction complex, and the relative light intensity (RLU) is measured. The hemoglobin concentration in the sample is proportional to the relative light intensity (RLU) measured by the optical system of the fully automatic chemiluminescence immunoassay. The instrument automatically fits and calculates the hemoglobin concentration.

[0077] (4) According to the interference model formula:

[0078] y=-0.8053x 2 +36.589x-4.3696, where x is the hemoglobin concentration in the hemolyzed serum, and y is the total concentration of NSE released by hemoglobin and erythrocytes in the hemolyzed serum;

[0079] Taking the hemoglobin concentration test result as x, substituting it into the interference model formula to obtain the y value;

[0080] The result obtained by subtracting the y value from the preliminary NSE concentration test result is the corrected NSE concentration test value.

[0081] Example 3

[0082] This embodiment provides the construction of an interference model.

[0083] 1. Preparation of interference curve

[0084] 1. Hemoglobin interferes with serum NSE

[0085] Test instrument: LunLuni2000 fully automatic chemiluminescence immunoassay analyzer produced by Shanghai Pinfeng Medical Technology Co., Ltd.

[0086] Experimental steps: After the pure hemoglobin is dissolved in a solvent, a series of gradient concentration samples are prepared. The series of gradient concentration samples are detected by the magnetic particle chemiluminescence kit prepared in Example 1. The results are shown in Table 1 and Figure 1 shown.

[0087] Table 1 Results of pure hemoglobin detection by magnetic microparticle chemiluminescence kit

[0088]

[0089] From Table 1 and Figure 1 It can be seen that hemoglobin will affect the measurement results and form a quadratic linear relationship (y = -0.3823x2 +11.43x+0.2183).

[0090] 2. Interference of NSE released from erythrocytes on serum NSE

[0091] A series of samples with known serum NSE content, hemolytic NSE content and hemoglobin content of the same person were selected as the calibration set.

[0092] The serum NSE detection value and hemolytic NSE detection value were detected by magnetic particle chemiluminescence kit, and the hemoglobin concentration value of the hemolytic sample was detected by hemoglobin magnetic particle chemiluminescence kit. The results are shown in Table 2.

[0093] Table 2 Results of serum NSE, hemolytic NSE and hemoglobin concentrations of the same person

[0094]

[0095]

[0096] Hemolytic NSE concentration = serum NSE concentration + hemoglobin NSE concentration + erythrocyte-released NSE concentration.

[0097] Therefore, the relationship between NSE released by red blood cells and hemoglobin is shown in Table 3 and Figure 2 shown.

[0098] Table 3 Hemoglobin and NSE concentration released by erythrocytes

[0099] Hemoglobin concentration (g / L) NSE concentration released by erythrocytes (ng / mL) 1.507 33.208 9.844 199.833 8.222 176.753 1.709 37.01 7.735 166.032 4.825 102.586 4.561 102.859 2.011 44.208 1.747 37.915 0.911 18.259

[0100] From Table 3 and Figure 2 It can be seen that the hemoglobin concentration and the NSE concentration released by red blood cells have a quadratic linear relationship (y = -0.423x 2 +25.159x-4.5879).

[0101] 3. Establishment of interference formula

[0102] Interference NSE = hemoglobin NSE + red blood cell release NSE. As mentioned above, both are linearly related to hemoglobin. Therefore, the interference formula can be established through the relationship between hemoglobin and the two, y = -0.8053x 2 +36.589x-4.3696, such as Figure 3 shown.

[0103] 2. Application of Interference Formula

[0104] Test instrument: LunLuni2000 fully automatic chemiluminescence immunoassay analyzer produced by Shanghai Pinfeng Medical Technology Co., Ltd.

[0105] Experimental steps: 5 NSE serum samples and hemolysis samples from the same person were selected and tested using the provided NSE magnetic particle chemiluminescence kit and the hemoglobin magnetic particle chemiluminescence kit (magnetic particle chemiluminescence kit prepared in Example 1). The concentration of the hemolysis sample was corrected by the interference formula and compared with the serum sample. The results are shown in Table 4.

[0106] Table 4 Comparison of serum sample concentration and corrected hemolysis sample concentration

[0107]

[0108] It can be seen from Table 4 that the concentration corrected by the interference formula is not much different from the serum sample concentration.

[0109] It can be seen that the NSE detection method and kit based on hemoglobin content back-calculation of the present invention can accurately evaluate the interference of hemoglobin in hemolyzed samples on NSE detection, and improve the accuracy of NSE detection of hemolyzed samples through hemoglobin content back-calculation correction.

[0110] Finally, it should be noted that the terms "comprises," "includes," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements that are inherent to such process, method, article, or apparatus.

[0111] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0112] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A magnetic microparticle chemiluminescence kit for NSE calibration detection of hemolytic samples, characterized in that: The kit comprises magnetic particles coated with NSE monoclonal antibodies, magnetic particles coated with hemoglobin monoclonal antibodies, acridinium ester-labeled NSE monoclonal antibodies, acridinium ester-labeled hemoglobin monoclonal antibodies, NSE series calibrators and hemoglobin series calibrators.

2. A magnetic microparticle chemiluminescence kit for NSE correction detection of hemolytic samples according to claim 1, characterized in that: In the NSE series of calibrators, the concentration range of NSE is 1 to 300 ng / mL; In the hemoglobin series calibrators, the hemoglobin concentration range is 0 to 200 g / L.

3. A magnetic microparticle chemiluminescence kit for NSE correction detection of hemolytic samples according to claim 1, characterized in that: The NSE monoclonal antibody-coated magnetic particles are suspended in a blocking storage solution to prepare a NSE monoclonal antibody-coated magnetic particle solution with a concentration of 0.1 to 0.4 μg / mL; The hemoglobin monoclonal antibody-coated magnetic particles are suspended in the closed storage solution to prepare a hemoglobin monoclonal antibody-coated magnetic particle solution with a concentration of 0.1 to 0.4 μg / mL; The acridinium ester-labeled NSE monoclonal antibody is dissolved in a labeling diluent to prepare an acridinium ester-labeled NSE monoclonal antibody solution with a concentration of 0.05 to 0.5 μg / mL; The acridinium ester-labeled hemoglobin monoclonal antibody is dissolved in the labeling diluent to prepare an acridinium ester-labeled hemoglobin monoclonal antibody solution with a concentration of 0.05 to 0.5 μg / mL; The closed preservation solution contains the following ingredients: Tris-HCl: 0.02~0.1M; Bovine serum albumin: 0.5~3wt%; ProClin300: 0.05~0.3wt%; Triton X100: 0.05~0.3wt%; Sodium chloride: 0.5~0.9wt%; The pH value of the sealing storage solution is 7 to 8; The labeling diluent contains the following ingredients: Tris-HCl: 0.02~0.1M; Bovine serum albumin: 0.5~3wt%; ProClin300: 0.05~0.3wt%; Triton X100: 0.05~0.3wt%; Magnesium chloride: 0.1~0.5wt%.

4. A magnetic microparticle chemiluminescence kit for NSE correction detection of hemolytic samples according to claim 1, characterized in that: The NSE monoclonal antibody-coated magnetic microparticles are prepared by covalently binding the NSE monoclonal antibody to the surface of the carboxyl-modified magnetic microparticles; The hemoglobin monoclonal antibody-coated magnetic particles are prepared by covalently binding the hemoglobin monoclonal antibody to the surface of the carboxyl-modified magnetic particles; The acridinium ester-labeled NSE monoclonal antibody and the acridinium ester-labeled hemoglobin monoclonal antibody are both prepared by dialysis; The NSE series calibrants are prepared by diluting NSE with a bovine serum albumin solution and a MgCl solution; The hemoglobin series calibrants are prepared by diluting hemoglobin with a bovine serum albumin solution and a MgCl solution.

5. The magnetic microparticle chemiluminescence kit for NSE correction detection of hemolytic samples according to claim 1, characterized in that: The NSE monoclonal antibody includes NSE mouse monoclonal antibody; The hemoglobin monoclonal antibody includes a human hemoglobin monoclonal antibody.

6. Use of the magnetic microparticle chemiluminescence kit for NSE correction detection of hemolyzed samples as claimed in any one of claims 1 to 5 in the detection of NSE content in hemolyzed samples for non-disease diagnosis purposes.

7. A NSE detection method based on back calculation of hemoglobin content, characterized in that: The detection method comprises: The NSE concentration of the hemolyzed sample was tested using a magnetic particle chemiluminescence kit to obtain preliminary NSE concentration test results; Using the magnetic particle chemiluminescence kit to detect the hemoglobin concentration of the hemolyzed sample to obtain a hemoglobin concentration detection result; According to the hemoglobin concentration detection result and the interference model, the preliminary NSE concentration detection result is back-calculated and corrected to obtain a corrected NSE concentration detection value; Wherein, the magnetic particle chemiluminescence kit comprises a magnetic particle chemiluminescence kit for NSE correction detection of hemolytic samples according to any one of claims 1-5.

8. The NSE detection method based on hemoglobin content back calculation according to claim 7, characterized in that: The method of detecting the NSE concentration of the hemolyzed sample using a magnetic particle chemiluminescence kit to obtain a preliminary detection result of the NSE concentration specifically includes: The NSE concentration of the hemolyzed sample was detected using magnetic particles coated with NSE monoclonal antibody and acridinium ester-labeled NSE monoclonal antibody in the magnetic particle chemiluminescence kit to obtain preliminary detection results of NSE concentration.

9. The NSE detection method based on hemoglobin content back calculation according to claim 7, characterized in that: The method of detecting the hemoglobin concentration of the hemolyzed sample using the magnetic particle chemiluminescence kit to obtain the hemoglobin concentration detection result specifically includes: The hemoglobin concentration of the hemolyzed sample is detected using the magnetic particles coated with the hemoglobin monoclonal antibody and the hemoglobin monoclonal antibody labeled with acridinium ester in the magnetic particle chemiluminescence kit to obtain the hemoglobin concentration detection result.

10. The NSE detection method based on hemoglobin content back calculation according to claim 7, characterized in that: The step of back-calculating and correcting the preliminary NSE concentration detection result according to the hemoglobin concentration detection result and the interference model to obtain a corrected NSE concentration detection value specifically includes: The interference model formula is as follows: y=-0.8053x 2 +36.589x-4.3696, where x is the hemoglobin concentration in the hemolyzed serum, and y is the total concentration of NSE released by hemoglobin and erythrocytes in the hemolyzed serum; Taking the hemoglobin concentration test result as x, substituting it into the interference model formula to obtain the y value; The result obtained by subtracting the y value from the preliminary NSE concentration test result is the corrected NSE concentration test value.

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