Anti-LKM (liver-kidney microsomal) 1 antibody detection kit and detection method

A technology for antibody detection and microsomes, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of unsuitable laboratories, cross-contamination of reagents, complicated operation, etc., and achieve shortened detection time, simple and convenient operation, good stability effect

Inactive Publication Date: 2016-02-17
SUZHOU HAOOUBO BIOPHARML
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] (1) False positives may occur using this method
[0010] (2) When analyzing the results, non-specific recognition cannot be distinguished according to the size of the molecular weight
[0011] (3) 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
[0012] (4) The background in the fluorescence measurement is high, and it is difficult to use the fluorescence immunoassay technique for quantitative determination
[0013] (5) Experienced professionals are required to judge the results, and the objectivity of the analysis results is insufficient
[0016] (1) The detection reagent is an open method during the detection process, which may easily cause cross-contamination between various reagents and affect the detection results
[0017] (2) ELISA methods mostly use horseradish peroxidase for detection, and its detection range and sensitivity are low
[0018] (3) The detection time of the ELISA method is usually relatively long, and the total time required to complete a test is generally more than 2 hours, which cannot fully meet the needs of clinical rapid diagnosis
[0019] (4) The ELISA method cannot be randomly injected for detection, and there is a lag in the detection results

Method used

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  • Anti-LKM (liver-kidney microsomal) 1 antibody detection kit and detection method

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Embodiment 1

[0034] Provides an anti-liver-kidney microsome 1 antibody detection kit, including biotinylated LKM-1 antigen (R1), calibrator, quality control, AP-anti-human IgG antibody (R2), magnetic particle reagent (M), Chemiluminescence substrate and wash solution. The biotinylated LKM-1 antigen is an LKM-1 antigen coupled with biotin. The AP-anti-human IgG antibody is an AP-labeled anti-human IgG antibody, in which alkaline phosphatase AP is coupled to an anti-human IgG antibody. The magnetic particle reagent is nano magnetic particle suspension with avidin SA group on the surface.

[0035] The biotinylated LKM-1 antigen is biotin and -NH on the surface of the LKM-1 antigen molecule 2 obtained by group reaction. The calibrator is prepared from human serum containing anti-LKM-1 antibody to obtain a calibration curve; the quality control product is prepared from human serum containing anti-LKM-1 antibody. The diameter of the magnetic particle in the magnetic particle reagent is 0.1-0...

Embodiment 2

[0038] Performance evaluation of the anti-liver-kidney microsome 1 antibody detection kit of the present invention:

[0039] (1) Sample comparison

[0040]Negative and positive coincidence rate: The kit of the present invention detects the content of anti-LKM-1 antibody in 217 cases of clinical serum, and conducts clinical comparison with similar products of well-known foreign companies. The comparison results are shown in the table below. The results showed that the positive coincidence rate of the anti-LKM-1 antibody kit of the present invention was 90.0% (45 / 50), and the negative coincidence rate was 99% (211 / 213), indicating that the clinical coincidence rate of the kit is high.

[0041]

[0042] (2) Sensitivity: The LOD of this detection kit is 0.145RU / mL, while the sensitivity of ELISA is 2RU / mL.

[0043] (3) Linearity: Use a kit to detect diluted samples according to 1 / 2, 1 / 4, 1 / 8, 1 / 16, 1 / 40, 1 / 80, 1 / 200, 1 / 400 of a high-value serum , make a regression curve with ...

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Abstract

The invention discloses an anti-LKM (liver-kidney microsomal) 1 antibody detection kit and a detection method. The kit comprises biotinylation LKM-1 antigen, a calibrator, a quality control material, an AP (alkaline phosphatase)-anti-human IgG (intravenous gamma globulin) antibody, a magnetic particle reagent, a chemiluminescent substrate and a cleaning liquid. According to the detection method, the reagent in the kit is used for detection of the sample. With the adoption of the scheme, a full-automatic magnetism particulate chemiluminescent detection method is adopted for selection of an alkaline phosphatase AP-adamantane AMPPD system, and the kit has the advantages of good stability, high sensitivity, good repeatability and the like; meanwhile, the detection time is greatly shortened, the total time required for one test is about 50 minutes, the operation is simple and convenient, and full-automatic detection is really realized.

Description

technical field [0001] The invention relates to the technical field of magnetic particle chemiluminescence immunodiagnosis, in particular to an anti-liver-kidney microsome 1 antibody detection kit and detection method. Background technique [0002] In recent years, the research on autoimmune liver disease at home and abroad has become more and more in-depth, and has progressed rapidly, involving pathogenesis, clinical diagnosis, clinical treatment, prognosis, genetics, and research on related autoantibodies and target antigens. [0003] Liver-kidney microsomal (LKM) autoantibodies are related to liver inflammatory diseases, and LKM autoantibodies are related to autoimmune chronic active hepatitis, also known as LKM-1 autoantibodies. LKM-1 is a protein with a molecular weight of 50KDa, which was first discovered in rat liver as cytochrome P450db1. Recent studies have reported that three P450 cytochromes ( A2, D6 and C9) can be used as an autoantigen for hepatitis disea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/576
CPCG01N33/53G01N33/576
Inventor 张伟柳乐秦枫崔利歌赵转王健王静马竹凤黄琪赵婷
Owner SUZHOU HAOOUBO BIOPHARML
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