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Kit for detecting anti-mutant citrullinated waveform protein antibody and detection method

A citrullination and vimentin technology, applied in the field of immunoassay, can solve the problems of complicated operation process, long experiment time, low accuracy and so on

Pending Publication Date: 2021-07-13
北京森美希克玛生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the enzyme-linked immunoassay method can accurately detect the concentration of anti-mutant citrullinated vimentin antibody in the sample, the actual operation takes a long time and the operation process is complicated
Colloidal gold method and immunofluorescence chromatography can be quickly detected, but only qualitative or semi-quantitative detection, the accuracy is low, and cannot provide a reliable basis for clinical judgment

Method used

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  • Kit for detecting anti-mutant citrullinated waveform protein antibody and detection method
  • Kit for detecting anti-mutant citrullinated waveform protein antibody and detection method
  • Kit for detecting anti-mutant citrullinated waveform protein antibody and detection method

Examples

Experimental program
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Effect test

Embodiment 1

[0039] This embodiment provides a kit for detecting anti-MCV antibodies, which includes: streptavidin-labeled magnetic particle solution, biotin-labeled mutant citrullinated vimentin solution, acridinium ester-labeled mouse anti-human antibody solutions, substrates compatible with acridinium esters, calibrators, controls, and sample diluents.

[0040] Wherein, the substrate includes excitation solution A and excitation solution B, and excitation solution A contains: 0.07mol / L HCl and 0.65wt% H 2 o 2 , pH 1.2; excitation solution B contains: 0.6mmol / L NaOH and 0.65wt% dodecyltrimethylammonium iodide, pH 13.2.

[0041] The above-mentioned streptavidin-labeled magnetic particle solution was prepared by the following method:

[0042] (1) Take amino-modified magnetic particles (particle size: 0.1-5 μm) for terminal modification and activation: take 50 mg of amino-modified magnetic particles in a beaker, add an equal volume of acetone and methanol 2 ml, wash 3 times, magnetically ...

Embodiment 2

[0057] This embodiment provides a method for detecting anti-MCV antibodies using the kit of Embodiment 1, which includes the following steps:

[0058] (1) When the test sample is collected serum or plasma (handle the sample to avoid contamination, the frozen sample must be completely thawed, and then fully mixed), draw 10 μl of the sample and dilute it 10 times with the sample diluent, and mix it for later use;

[0059] (2) Take 50 μl of the diluted sample, add 50 μl of streptavidin-labeled magnetic particle solution, 50 μl of biotin-labeled mutant citrullinated vimentin solution, and 50 μl of acridinium ester-labeled mouse anti-human antibody solution, Incubate at 37°C for 18 minutes.

[0060] (3) Wash 3 times with washing solution after incubation, the washing solution is sodium chloride containing 2mol / L, 2wt% Tween-20 and 0.1mol / L-1mol / L phosphate buffer, buffer The pH is 7.8.

[0061] (4) After washing, add 100 μl of pre-excitation solution A, and then add 100 μl of exc...

Embodiment 3

[0063] The composition of the kit for detecting anti-MCV antibodies provided in this example is basically the same as that in Example 1, the difference is that this example uses alkaline phosphatase-labeled mouse anti-human antibody as the labeled antibody, corresponding to the chemiluminescence substrate used Be (adamantane)-1,2-dioxetane (AMPPD); 7.00-9.50), 0.2mol / L sodium sulfate, 0.1mol / L sodium chloride, 0.2wt% Proclin-300 and 2wt% luminescence enhancer.

[0064] The preparation method of the above-mentioned alkaline phosphatase-labeled mouse anti-human antibody is as follows:

[0065] Weigh 25 mg of alkaline phosphatase in glutaraldehyde solution, mix thoroughly, and place it at room temperature overnight. The reacted solution is eluted with sodium carbonate solution (0.2mol / L, pH 9.0) through a chromatographic column. Collect the effluent liquid, dissolve 12.5mg of mouse anti-human antibody in sodium carbonate solution (0.2mol / L, pH 9.0), stir until completely dissolv...

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Abstract

The invention discloses a kit for detecting an anti-mutant citrullinated waveform protein antibody and a detection method, and relates to the technical field of immunoassay. The kit disclosed by the invention comprises magnetic particles labeled by streptavidin, mutant citrullinated waveform protein labeled by biotin, a labeled antibody labeled by a detectable marker, and a substrate applicable to the detectable marker. The kit has the characteristics of high sensitivity, good specificity, wide linear range and the like, and can quickly and accurately detect the anti-mutant citrullinated waveform protein antibody in a blood sample.

Description

technical field [0001] The invention relates to the technical field of immune analysis, in particular to a kit and a detection method for detecting anti-mutant citrullinated vimentin antibodies. Background technique [0002] Rheumatoid arthritis (RA) is a disabling chronic polyarthritis disease. It is characterized by chronic inflammation of synovial joints leading to physical disability, and the incidence is also affected by factors such as genetics and environment, and the incidence rate has a tendency to increase year by year. Some RA patients can experience irreversible bone and joint destruction within 2 years of onset, resulting in joint deformity and loss of exercise capacity. Studies have found that the earlier the intervention and treatment of RA, the more effective it is to control the deterioration of the disease and reduce the impact on the body. Therefore, early diagnosis and treatment are very important. [0003] The most common markers of RA are: anti-cyclic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/58G01N33/543
CPCG01N33/6854G01N33/6893G01N33/54326G01N33/581G01N33/583G01N2800/102
Inventor 肖文峰庞启锋胡国茂
Owner 北京森美希克玛生物科技有限公司
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