CN107807238A - Anticardiolipin antibodies chemiluminescence immune detection reagent kit and preparation method thereof - Google Patents
Anticardiolipin antibodies chemiluminescence immune detection reagent kit and preparation method thereof Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- 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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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
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Abstract
The invention discloses a kind of anticardiolipin antibodies chemiluminescence immune detection reagent kit and preparation method thereof, anticardiolipin antibodies chemiluminescence immune detection reagent kit includes：The chemiluminescent labels of magnetic particle and human immunoglobulins' secondary antibody mark of the coated carboxylated of anticardiolipin antibodies monoclonal antibody.This anticardiolipin antibodies chemiluminescence immune detection reagent kit can be using Full-automatic chemiluminescence immunoassay analysis meter as detection instrument, complete this anticardiolipin antibodies chemiluminescence immune detection reagent kit of detection of anticardiolipin antibodies, by experiment, its detection sensitivity reaches 0.2AU/mL, 10 times are at least improved relative to the detection method sensitivity of traditional anticardiolipin antibodies, the accuracy of detection of this anticardiolipin antibodies chemiluminescence immune detection reagent kit is higher.
Description
Technical Field
The invention relates to the field of in-vitro detection, in particular to a chemiluminescence immunoassay kit for an anticardiolipin antibody and a preparation method thereof.
Background
Anti-cardiolipin Antibodies (ACLs) are autoantibodies that use negatively charged cardiolipin on platelet and endothelial cell membranes as target antigens.
Cardiolipin is an antigenic phospholipid isolated from the bovine myocardium by pancborn in 1941 and is named, and is the highest in mammalian muscle, and contains 4 unsaturated fatty acids per cardiolipin molecule, which is highly susceptible to oxidation. Anti-cardiolipin antibodies are common in Systemic Lupus Erythematosus (SLE) and other autoimmune diseases. The antibody can be used for treating thrombosis, small blood plate, spontaneous abortion, or intrauterine stillbirth. Studies have demonstrated that many factors are closely related to ACL production, common causes being:
(1) autoimmune diseases such as Systemic Lupus Erythematosus (SLE), Rheumatoid Arthritis (RA) scleroderma, etc.;
(2) viral infections such as adenovirus, rubella virus, varicella virus, mumps virus, etc.;
(3) other diseases such as mycoplasma system diseases;
(4) some drugs, such as chlorpromazine, phenothiazine, etc., are taken orally.
The main method for clinically detecting the anticardiolipin antibody is an enzyme-linked immunosorbent assay, but the method has the following defects:
(1) the special 96-hole micro-porous plate of 12 multiplied by 8 type, 6 multiplied by 8 type, 8 multiplied by 12 type or whole plate type is used as an antigen coating tool and a reaction vessel, and can be used only once by being divided into 12 batches, 6 batches, 8 batches or whole plate when in use, and the independent detection of single person can not be carried out;
(2) the quantitative determination has more types of reagents, each detection reagent is contained in a reagent bottle, and each reagent needs to be respectively filled into the micropores of the microporous plate by replacing a liquid suction nozzle when one reagent is used, so that not only are the types of the reagent bottles multiple, but also the operation of filling the reagents is extremely complicated;
(3) the corresponding label of the detection information is lacked, the production batch number and the validity period information of the detection reagent can be known or known only by looking up the mark of the external packing box of the kit, and the known information is not controlled in the detection process and has great randomness;
(4) the detection reagent is in an open space in the detection process, so that cross contamination among various reagents is easily caused to influence the accuracy of a detection result;
(5) manual operation is mostly adopted in the detection process, the addition of a reagent or a sample is not very accurate, the operation process is extremely complicated and complicated, operation errors are easy to occur, and the accuracy and precision of a detection result are poor;
(6) the quantity of the reagent kit in the detection items is multiplied by 48/96, if 10 items need to be detected, the quantity of the reagent kit needs to be 10 multiplied by 48/96, if only one sample needs to be detected for 10 different items, the reagent kit needs to be also multiplied by 10 multiplied by 48/96, and the method has the defect of being not economic and reasonable.
Disclosure of Invention
Therefore, it is necessary to provide a chemiluminescence immunoassay kit for an anticardiolipin antibody with higher detection sensitivity and a preparation method thereof.
A chemiluminescence immunoassay kit for an anti-cardiolipin antibody comprises: anti-cardiolipin antibody monoclonal antibody coated carboxylated magnetic microparticles and anti-human immunoglobulin secondary antibody labeled chemiluminescent markers.
In one embodiment, in the carboxylated magnetic microparticles coated with the anti-cardiolipin antibody monoclonal antibody, the ratio of the anti-cardiolipin antibody monoclonal antibody to the carboxylated magnetic microparticles is 1: 25 to 35.
In one embodiment, in the anti-human immunoglobulin secondary antibody labeled chemiluminescent label, the ratio of the anti-cardiolipin antibody monoclonal antibody to the chemiluminescent label is 50: 1 to 10.
In one embodiment, the carboxylated magnetic particles have a particle size of 0.05 μm to 1 μm.
In one embodiment, the chemiluminescent label is luminol, isoluminol, ruthenium terpyridyl, or acridinium ester.
In one embodiment, the chemiluminescent substrate solution comprises a solution A and a solution B.
In one embodiment, the solution A is H2O2And the solution B is NaOH solution.
In one embodiment, an anti-cardiolipin antibody calibrator is also included.
In one embodiment, the anticardiolipin antibody calibrator is a solution of anticardiolipin antibody at a concentration of 0AU/mL, 10AU/mL, 20AU/mL, 50AU/mL, 100AU/mL and 200AU/mL, respectively.
The preparation method of the chemiluminescence immunoassay kit for the anticardiolipin antibody comprises the following steps:
taking a suspension of the carboxylated magnetic particles, carrying out magnetic separation to remove a supernatant, then carrying out resuspension by using an MES buffer solution, then adding an EDC aqueous solution, activating surface carboxyl of the carboxylated magnetic particles, then adding an anticardiolipin antibody monoclonal antibody, suspending for 2-10 h at room temperature, carrying out magnetic separation to remove the supernatant, and then carrying out resuspension by using a Tris buffer solution to obtain carboxylated magnetic particles coated by the anticardiolipin antibody monoclonal antibody; and
and adding the anti-cardiolipin antibody monoclonal antibody into a carbonate buffer solution, uniformly mixing, adding a chemiluminescent marker, uniformly mixing, reacting at room temperature in a dark place for 1-2 h, and removing impurities to obtain the anti-human immunoglobulin secondary antibody labeled chemiluminescent marker.
The chemiluminescence immunoassay kit for the anticardiolipin antibody can complete the detection of the anticardiolipin antibody by taking a full-automatic chemiluminescence immunoassay analyzer as a detection tool, and through experiments, the detection sensitivity of the chemiluminescence immunoassay kit for the anticardiolipin antibody reaches 0.2AU/mL, is at least 10 times higher than that of the traditional detection method for the anticardiolipin antibody, and has higher detection precision.
Drawings
FIG. 1 is a flow chart illustrating a method for preparing a chemiluminescent immunoassay kit for an anti-cardiolipin antibody according to one embodiment;
FIG. 2 is a graph showing the standard curve of the anti-cardiolipin antibody obtained in example 3.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in detail below. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein, but rather should be construed as broadly as the present invention is capable of modification in various respects, all without departing from the spirit and scope of the present invention.
The chemiluminescent immunoassay kit for an anti-cardiolipin antibody of one embodiment comprises: anti-cardiolipin antibody monoclonal antibody coated carboxylated magnetic microparticles and anti-human immunoglobulin secondary antibody labeled chemiluminescent markers.
Preferably, in the carboxylated magnetic microparticles coated with the anti-cardiolipin antibody monoclonal antibody, the ratio of the anti-cardiolipin antibody monoclonal antibody to the carboxylated magnetic microparticles is 1: 25 to 35.
Preferably, in the anti-human immunoglobulin secondary antibody labeled chemiluminescent marker, the ratio of the anti-cardiolipin antibody monoclonal antibody to the chemiluminescent marker is 50: 1 to 10.
Preferably, the carboxylated magnetic microparticles have a particle size of 0.05 to 1 μm.
The chemiluminescent label may be luminol, isoluminol, ruthenium terpyridyl or acridinium ester. Among them, the chemiluminescent label is preferably an acridinium ester.
In other embodiments, the chemiluminescent immunoassay kit for the anti-cardiolipin antibody further comprises a chemiluminescent substrate solution.
The chemiluminescent substrate solution comprises solution A and solution B. The solution A can be H2O2Solution B can be NaOH solution.
In this example, the solution A was H with a concentration of 0.1mol/L2O2The solution B is NaOH solution with the concentration of 0.25 mol/L.
In other embodiments, the anti-cardiolipin antibody chemiluminescent immunoassay kit further comprises an anti-cardiolipin antibody calibrator.
The anticardiolipin antibody calibrator is a solution of anticardiolipin antibody at a concentration of 0AU/mL, 10AU/mL, 20AU/mL, 50AU/mL, 100AU/mL and 200AU/mL, respectively.
Specifically, the anticardiolipin antibody calibrator may employ a standard buffer to formulate the anticardiolipin antibody into solutions having concentrations of 0AU/mL, 10AU/mL, 20AU/mL, 50AU/mL, 100AU/mL and 200AU/mL, respectively.
When the anti-cardiolipin antibody chemiluminescence immunoassay kit is used for detecting an anti-cardiolipin antibody, a full-automatic chemiluminescence immunoassay analyzer is used for detecting an anti-cardiolipin antibody calibration sample, a standard curve is drawn, and the kit is arranged in computer software; then testing an actual sample, and calculating the concentration of the sample according to the luminous value of the sample; and finally, evaluating the performance (sensitivity, linearity, precision and interference) of the full-automatic chemiluminescence immunoassay system for the anti-cardiolipin antibody.
The chemiluminescence immunoassay kit for the anticardiolipin antibody can complete the detection of the anticardiolipin antibody by taking a full-automatic chemiluminescence immunoassay analyzer as a detection tool, and through experiments, the detection sensitivity of the chemiluminescence immunoassay kit for the anticardiolipin antibody reaches 0.2AU/mL, is at least 10 times higher than that of the traditional detection method for the anticardiolipin antibody, and has higher detection precision.
In addition, the chemiluminescence immunoassay kit for the anticardiolipin antibody also has the following advantages:
1. acridinium ester is selected as a marker material and is applied to a chemiluminescence immunoassay system, the luminophore system is direct chemiluminescence, and compared with the traditional enzymatic chemiluminescence, the reaction does not need enzyme participation, so that the cost is saved;
2. the chemiluminescence immunoassay system of the acridinium ester has wide linear range which can reach 2 AU/mL-120 AU/mL, while the traditional detection method of the anticardiolipin antibody has the linear detection range of 10 AU/mL-120 AU/mL;
3. the acridinium ester chemiluminescence immunoassay system has high repeatability, and the difference between batches and between batches is within 5 percent, which is difficult to achieve by other chemiluminescence immunoassay systems;
4. the chemiluminescence immune analysis system realizes the quantification of the sample, and the concentration value of the sample can be directly obtained only by testing the sample through internally arranging a standard curve to test software;
5. the chemiluminescence immunoassay system can realize full automation, the reagent and sample are added by instruments, the operation is simpler and more convenient, and the artificial error is reduced.
The preparation method of the anti-cardiolipin antibody chemiluminescence immunoassay kit shown in figure 1 comprises the following steps:
taking a suspension of the carboxylated magnetic particles, carrying out magnetic separation to remove a supernatant, then carrying out resuspension by using an MES buffer solution, then adding an EDC aqueous solution, activating surface carboxyl of the carboxylated magnetic particles, then adding an anticardiolipin antibody monoclonal antibody, suspending for 2-10 h at room temperature, carrying out magnetic separation to remove the supernatant, and then carrying out resuspension by using a Tris buffer solution to obtain the carboxylated magnetic particles coated by the anticardiolipin antibody monoclonal antibody.
MES (2- (N-morpholino) ethanesulfonic acid) buffer at 0.02M pH 5.5.
Tris buffer was 0.1M in concentration and contained 2% BSA, pH 8.0.
The concentration of the EDC (1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide) aqueous solution is 10mg/mL to 20mg/mL, and the ratio of EDC to carboxylated magnetic microparticles is 0.05: 0.1 to 1.
Preferably, in the carboxylated magnetic microparticles coated with the anti-cardiolipin antibody monoclonal antibody, the ratio of the anti-cardiolipin antibody monoclonal antibody to the carboxylated magnetic microparticles is 1: 25 to 35.
Preferably, the carboxylated magnetic microparticles have a particle size of 0.05 to 1 μm.
And adding the anti-cardiolipin antibody monoclonal antibody into a carbonate buffer solution, uniformly mixing, adding a chemiluminescent marker, uniformly mixing, reacting at room temperature in a dark place for 1-2 h, and removing impurities to obtain the anti-human immunoglobulin secondary antibody labeled chemiluminescent marker.
The concentration of the carbonate buffer solution is 0.1M, the pH value is 9.0-9.5,
the operation of impurity removal is centrifugal desalting column desalination, and the specific operation is as follows: the centrifugation desalting column was treated with purified water and TBS buffer (40 mM Tris-HCl, 0.5% BSA, 1% NaCl, pH 8.0), respectively, and the resultant solution of carboxylated magnetic microparticles coated with the anti-cardiolipin antibody monoclonal antibody was added, and the liquid in the centrifuge tube was collected.
Preferably, in the anti-human immunoglobulin secondary antibody labeled chemiluminescent marker, the ratio of the anti-cardiolipin antibody monoclonal antibody to the chemiluminescent marker is 50: 1 to 10.
The chemiluminescent label may be luminol, isoluminol, ruthenium terpyridyl or acridinium ester. Among them, the chemiluminescent label is preferably an acridinium ester.
The obtained carboxylated magnetic particles coated by the anti-cardiolipin antibody monoclonal antibody and the anti-human immunoglobulin secondary antibody labeled chemiluminescent marker are combined to obtain the anti-cardiolipin antibody chemiluminescent immunoassay kit.
When the chemiluminescence immunoassay kit for the anticardiolipin antibody is used, chemiluminescence substrate liquid and an anticardiolipin antibody calibration product are also needed.
The chemiluminescent substrate solution and the anticardiolipin antibody calibration sample can be prepared by self.
The chemiluminescent substrate solution comprises solution A and solution B. The solution A can be H2O2Solution B can be NaOH solution.
In this example, the solution A was H with a concentration of 0.1mol/L2O2The solution B is NaOH solution with the concentration of 0.25 mol/L.
Specifically, the anticardiolipin antibody calibrator may employ a standard buffer to formulate the anticardiolipin antibody into solutions having concentrations of 0AU/mL, 10AU/mL, 20AU/mL, 50AU/mL, 100AU/mL and 200AU/mL, respectively.
The preparation method of the anti-cardiolipin antibody chemiluminescence immunoassay kit is simple and convenient, and the prepared anti-cardiolipin antibody chemiluminescence immunoassay kit has high detection sensitivity and good application prospect.
The following are specific examples.
Example 1: preparation of chemiluminescence immunoassay kit for anticardiolipin antibody
(1) Preparation of carboxylated magnetic microparticles coated with anti-cardiolipin antibody monoclonal antibodies:
taking 50mg of carboxylated magnetic particle (MagnaBind 21353) suspension with the particle size of 0.05-1 μ M, carrying out magnetic separation to remove the supernatant, carrying out resuspension by using 0.02M of buffer solution with the pH value of 5.5 MES, adding 1mL of newly configured 10mg/mL EDC aqueous solution, activating the carboxyl groups on the surface of magnetic beads, adding 4mg of anti-cardiolipin antibody monoclonal antibody (biorbyt, product number orb 48780), suspending for 6h at room temperature, carrying out magnetic separation, removing the supernatant, carrying out resuspension to 1mg/mL by using 0.1M of buffer solution with the pH value of 8.0 and containing 2% BSA to obtain carboxylated magnetic particles coated by the anti-cardiolipin antibody monoclonal antibody, and subpackaging 5mL of each bottle at 4 ℃ for later use.
(2) Preparation of anti-human immunoglobulin secondary antibody-labeled acridinium ester:
taking 50 mu L of an anti-cardiolipin antibody monoclonal antibody with the concentration of 25mg/mL, adding 150 mu L of carbonate buffer solution with the concentration of 0.1M, pH of 9.0-9.5, uniformly mixing, then adding 1.5 mu L of acridine ester solution with the concentration of 5mg/mL, uniformly mixing, carrying out light-shielding reaction at room temperature, taking out after 1.5h, desalting by using a 2mL zeba centrifugal desalting column, firstly respectively treating by using purified water and TBS buffer solution in the desalting process, finally adding the obtained anti-human immunoglobulin secondary antibody labeled acridine ester solution, collecting liquid in a centrifuge tube to a storage tube to obtain the anti-human immunoglobulin secondary antibody labeled acridine ester, and subpackaging 5mL of each bottle at 4 ℃ for later use.
(3) Preparation of anticardiolipin antibody calibration sample:
the anticardiolipin antibody was formulated with standard buffer (40 mM Tris-HCl, 0.5% BSA, 1% NaCl, pH 8.0) at concentrations of 0AU/mL, 10AU/mL, 20AU/mL, 50AU/mL, 100AU/mL and 200AU/mL, 0.5 mL per vial was lyophilized and stored at 4 ℃ for further use.
Example 2: chemiluminescence immune detection method for anticardiolipin antibody
The method adopts a double-antibody sandwich method by using a full-automatic chemiluminescence immunoassay analyzer (YHLO, product number iFlash 3000) as a detection tool, namely adding 50 mu L of sample, 50 mu L of carboxylated magnetic particles coated by the anticardiolipin antibody monoclonal antibody and 50 mu L of sampleReacting the anticardiolipin antibody treating solution for 20 min, adding 50 μ L of acridinium ester coated with anticardiolipin antibody, reacting for 20 min, performing magnetic separation, sending the reaction mixture into a dark room, and sequentially adding luminescent substrate A solution (H)2O2) And B liquid (NaOH) to carry out luminescence reaction, and finally, recording the luminescence value.
Example 3: performance evaluation of anti-cardiolipin antibody chemiluminescence immunoassay kit
The method of example 2 was used to test the anticardiolipin antibody calibrator and the standard curve was plotted as shown in FIG. 2.
Then, for the actual sample to be tested, the sample concentration is calculated according to the sample luminescence value.
Detection of sensitivity:
the sensitivity of the chemiluminescence immunoassay kit for the anticardiolipin antibody is calculated according to the recommended experimental scheme of CLSI EP17-A document, and the calculated sensitivity is 0.2 AU/mL.
And (3) linear detection:
and (3) carrying out linear analysis on the standard substances with the concentrations of 2AU/mL, 10AU/mL, 20AU/mL, 50AU/mL and 120AU/mL, calculating a linear correlation coefficient, wherein r =0.9996, and in addition, the linear range of the detection of the kit on the anticardiolipin antibody sample is 2 AU/mL-120 AU/mL.
And (3) measuring precision:
taking two anti-cardiolipin antibody samples with the concentrations of 10AU/mL and 100AU/mL, respectively carrying out 3 parallels on each concentration of each sample, detecting by using three batches of kits, and calculating the intra-batch and inter-batch differences of the kits, wherein the results show that the intra-batch and inter-batch differences of the kits are both less than 5%.
Interference experiments:
taking mixed serum and adding interferents respectively comprises the following steps: combining bilirubin, free bilirubin, hemoglobin, ascorbic acid and glyceride, wherein the adding proportion is 1: 20, the measurement values of the mixed serum and the mixed serum added with various interferents were measured, and the deviation between the two was calculated to be within an acceptable range of. + -. 10%. The result shows that the interference reaches the file standard of NCCLS, and can be used for accurately evaluating the condition of the anti-cardiolipin antibody in a clinical laboratory.
Example 4 comparative experiment of anti-cardiolipin antibody chemiluminescence immunoassay kit
The chemiluminescence detection method and the traditional enzyme-linked immunosorbent assay are respectively used for detecting anticardiolipin antibody samples with the concentrations of 0.1AU/mL and 20AU/mL, the detection sensitivity of the two methods is compared, and the data are shown in the following table:
as can be seen from the above table, the sensitivity of the chemiluminescence detection method is improved by about 20 times compared with that of the enzyme-linked immunosorbent assay.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A chemiluminescence immunoassay kit for an anti-cardiolipin antibody is characterized by comprising: anti-cardiolipin antibody monoclonal antibody coated carboxylated magnetic microparticles and anti-human immunoglobulin secondary antibody labeled chemiluminescent markers.
2. The chemiluminescent immunoassay kit for anti-cardiolipin antibody of claim 1, wherein the ratio of the anti-cardiolipin antibody monoclonal antibody to the carboxylated magnetic microparticles in the carboxylated magnetic microparticles coated with the anti-cardiolipin antibody monoclonal antibody is 1: 25 to 35.
3. The anti-cardiolipin antibody chemiluminescent immunoassay kit of claim 1, wherein the ratio of the anti-cardiolipin antibody monoclonal antibody to the chemiluminescent label in the anti-human immunoglobulin secondary antibody labeled chemiluminescent label is 50: 1 to 10.
4. The chemiluminescent immunoassay kit for an anti-cardiolipin antibody according to claim 1, wherein the carboxylated magnetic microparticles have a particle size of 0.05 μm to 1 μm.
5. The chemiluminescent immunoassay kit for an anti-cardiolipin antibody of claim 1, wherein the chemiluminescent label is luminol, isoluminol, ruthenium terpyridyl, or acridinium ester.
6. The chemiluminescent immunoassay kit for an anti-cardiolipin antibody according to claim 1, further comprising a chemiluminescent substrate solution, wherein the chemiluminescent substrate solution comprises solution A and solution B.
7. The chemiluminescent immunoassay kit for an anti-cardiolipin antibody of claim 6, wherein the solution A is H2O2And the solution B is NaOH solution.
8. The chemiluminescent immunoassay kit for anti-cardiolipin antibody of claim 1, further comprising an anti-cardiolipin antibody calibrator.
9. The anti-cardiolipin antibody chemiluminescent immunoassay kit of claim 8, wherein the anti-cardiolipin antibody calibrator is a solution of anti-cardiolipin antibody at a concentration of 0AU/mL, 10AU/mL, 20AU/mL, 50AU/mL, 100AU/mL, and 200AU/mL, respectively.
10. A method for preparing the anti-cardiolipin antibody chemiluminescent immunoassay kit according to any one of claims 1-9, comprising the steps of:
taking a suspension of the carboxylated magnetic particles, carrying out magnetic separation to remove a supernatant, then carrying out resuspension by using an MES buffer solution, then adding an EDC aqueous solution, activating surface carboxyl of the carboxylated magnetic particles, then adding an anticardiolipin antibody monoclonal antibody, suspending for 2-10 h at room temperature, carrying out magnetic separation to remove the supernatant, and then carrying out resuspension by using a Tris buffer solution to obtain carboxylated magnetic particles coated by the anticardiolipin antibody monoclonal antibody; and adding the anti-cardiolipin antibody monoclonal antibody into a carbonate buffer solution, uniformly mixing, adding a chemiluminescent marker, uniformly mixing, reacting at room temperature in a dark place for 1-2 h, and removing impurities to obtain the anti-human immunoglobulin secondary antibody labeled chemiluminescent marker.
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