Antinuclear antibody chemiluminescence immunoassay kit and preparation method thereof

Abstract

The invention discloses an antinuclear antibody chemiluminescence immunoassay kit and a preparation method thereof. The antinuclear antibody chemiluminescence immunoassay kit comprises antinuclear antibody monoclonal antibody-coated chloracetylated magnetic particles and an inhibin monoclonal antibody-labelled chemiluminescent marker. With a full-automatic chemiluminescence immunity analyzer as a testing tool, the antinuclear antibody chemiluminescence immunoassay kit can complete detection of antinuclear antibodies. Through experiments, detection sensitivity of the antinuclear antibody chemiluminescence immunoassay kit reaches 4 AU / mL. In comparison with sensitivity of a traditional antinuclear antibody detection method, the sensitivity of the invention is raised by at least 10 times. Detection precision of the antinuclear antibody chemiluminescence immunoassay kit is high.

Description

technical field [0001] The invention relates to the field of in vitro detection, in particular to an antinuclear antibody chemiluminescence immunoassay kit and a preparation method thereof. Background technique [0002] Antinuclear antibody (antinuclear antibody, ANA), also known as anti-nucleic acid antigen antibody, is a group of various components of eukaryotic cells deoxyribonucleoprotein (DNP), DNA, extractable nuclear antigen (ENA) and RNA as The general term for autoantibodies to target antigens, mainly present in serum. Antinuclear antibodies have varying degrees of positive rates in various autoimmune diseases, such as systemic lupus erythematosus (SLE, 95% to 100%), rheumatoid arthritis (RA, 10% to 20%), mixed Connective tissue disease (MCTD, 80%-100%), Sjogren's syndrome (SS, 10%-40%), systemic scleroderma (85%-90%), lupus hepatitis (95%-100%), Primary biliary cirrhosis (95% to 100%), etc. [0003] Common methods for clinical detection of antinuclear antibodies...

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Problems solved by technology

[0006] (1) Non-specific recognition cannot be distinguished according to the size of the molecular weight when analyzing the results;

[0007] (2) The operation is relatively complicated and requires a more expensive fluorescence microscope, which is difficult to promote in many primary hospitals and is not suitable for laboratories with a large number of specimens;

[0008] (3) The background in the fluorescence measurement is relatively high, and it is difficult to use the fluorescence immunoassay technique for quantitative determination;

[0009] (4) The judgment of the results requires experienced professionals, and the objectivity of the analysis results is insufficient;

[0010] (5) Only qualitative detection can be carried out, but quantitative determination cannot be carried out

[0013] (1) Use 12×8 type, 6×8 type, 8×12 type or whole plate type 96-well special microwell plate as the antigen coating equipment and reaction container, which can only be divided into 12 batches and 6 batches when used , 8 batches or the whole board can be used at one time, and independent and single-person testing cannot be carried out;

[0014] (2) There are many kinds of reagents used in quantitative determination, and each detection reagent must be contained in a reagent bottle, and each time a reagent is used, the suction nozzle needs to be replaced to fill the microwells of the microwell plate respectively , not only there are many types of reagent bottles, but also the operation of filling reagents is extremely cumbersome;

[0015] (3) There is a lack of corresponding labeling of the testing information. The production batch number and expiry date information of the testing reagent can only be known or known by checking the label on the outer packaging box of the kit, and the known information is not controlled during the testing process, which has great potential. large randomness;

[0016] (4) The detection reagents are in an open space during the detection process, which may easily cause cross-contamination among various reagents and affect the accuracy of the detection results;

[0017] (5) Manual operation is mostly used in the detection process, the addition of reagents or samples is not very precise, the operation process is extremely cumbersome and complicated, and operation errors are prone to occur, and the accuracy and precision of the detection results are poor;

[0018] (6) The quantity configuration and use of the complete set of reagents for the test items are the number of items × 48 / 96 persons. If 10 items need to be tested, the configuration and use of reagents must be 10×48 / 96 persons. If Only one sample needs to detect 10 different items, and it also needs to configure reagents for 10×48 / 96 people, which has the disadvantage of not being economical and reasonable

[0021] Enzymatic chemiluminescence mainly includes horseradish peroxidase (HRP) and alkaline phosphatase, but both have certain limitations. The main disadvantage of horseradish peroxidase is: In the presence of biomolecules, they will also be oxidized by H2O2 to emit light. The background is relatively high, which affects the signal-to-noise ratio. The reaction kinetics is complicated, and there are many influencing factors. easy

The main disadvantages of alkaline phosphatase are: it takes a long time for the substrate to reach the plateau, and the cost of the substrate is high, resulting in high detection cost and heavy burden on patients

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