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Cyclic citrullinated peptide antibody, rheumatoid factor detection kit, and preparation method thereof

A technology of rheumatoid factor and cyclic citrullinated peptide, which can be used in biological tests, measuring devices, material inspection products, etc., can solve the problems of strong interference and high intensity, and achieve the effects of high sensitivity, fast response and easy operation.

Inactive Publication Date: 2018-03-02
WEIHAI NEOPROBIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the usual fluorescence measurement, because the test sample contains a variety of fluorescent components, the background fluorescence (scattered light caused by colloidal particles and solvent molecules in the sample and non-specific fluorescence emitted by proteins and other compounds in the serum) has a large intensity and interference. Strong, become the bottleneck of large-scale promotion of fluorescence analysis

Method used

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  • Cyclic citrullinated peptide antibody, rheumatoid factor detection kit, and preparation method thereof
  • Cyclic citrullinated peptide antibody, rheumatoid factor detection kit, and preparation method thereof
  • Cyclic citrullinated peptide antibody, rheumatoid factor detection kit, and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The various components of the test paper card in the cyclic citrullinated peptide antibody and rheumatoid factor detection kit can be prepared by the following measures:

[0036] 1. Preparation of sample pad 2:

[0037] Soak the glass fiber membrane in the treatment solution containing 2.0% Triton X-100, 2% BSA, 0.1M Tris buffer, pH7.5, soak at 4°C for 4 hours, then place it in an oven, and dry it at 37°C 2 hours.

[0038] 2. Preparation of binding pad 3 for absorbing fluorescent microsphere-labeled antibody:

[0039] Soak the glass fiber membrane in 200mM Tris-HCL treatment solution (containing 1.5% Triton X-100, 1.5% BSA, pH7.5), soak at 4°C for 4 hours, then take it out of a 37°C oven and dry it for 4 hours, and set it aside. The glass fiber membrane was placed on the Bio-DotXYZ3050 three-dimensional spraying platform, and the rare earth fluorescent microsphere-labeled cyclic citrullinated peptide antibody and rheumatoid factor monoclonal antibody were sprayed onto...

Embodiment 2

[0048] Embodiment 2: accuracy test

[0049] Select the above-mentioned test paper card and fluorescent immunochromatography analyzer (model: NEO-007), the setting of the parameters of the fluorescent immune analyzer: after setting the process parameters of the test paper card on the fluorescent immune analyzer, take the above-mentioned assembled test paper card , use 0.2, 0.5, 1, 2, 5, 20ng / ml cyclic citrullinated peptide antibody and rheumatoid factor calibrator respectively, measure with test paper card, get the fluorescence intensity value of each calibrator, and input the result into the analysis In the parameters of the analyzer, complete the setting of the parameters of the analyzer.

[0050] Main testing materials: clinical samples obtained from relevant hospitals, a total of 200 Roche electrochemiluminescence immunoassay value samples, including 100 serum samples, 100 whole blood samples, cyclic citrullinated peptide antibody and rheumatoid factor content distribution ...

Embodiment 3

[0059] Embodiment 3: precision test

[0060] Using the test paper card and measuring system of Example 2, the test paper card and the fluorescent immunochromatographic analyzer of the present invention were tested for precision.

[0061]Main testing materials: clinical samples were obtained from relevant hospitals, a total of 2 serum samples with chemiluminescence immunoassay value, among which the clinical measurement value of the low value fixed value sample was 0.24ng / ml, and the clinical measurement value of the high value fixed value sample was 1.78ng / ml .

[0062] Preparation:

[0063] Using the test paper card and measuring system of Example 2, each of the 2 fixed-value samples was repeatedly measured 20 times.

[0064] Analysis of test results:

[0065] After the clinical sample testing reagents are prepared, the clinical samples are tested according to the preparation method, and the test results are analyzed.

[0066] test results:

[0067] The low-value definit...

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Abstract

The invention relates to the technical field of fluorescent immunochromatography in medical immunology, and particularly provides a cyclic citrullinated peptide antibody, a rheumatoid factor detectionkit, and a preparation method thereof. The kit is provided with a test paper card, which successively includes a PVC board, a sample pad, a combining pad, a nitrocellulose membrane and a water absorbing pad from bottom to top. Rare earth fluorescent microsphere labeled cyclic citrullinated peptide antibody and rheumatoid factor monoclonal antibody are adsorbed on the combining pad, wherein diameters of the rare earth fluorescent microspheres are 60-120 nm, and the rare earth fluorescent microspheres are doped with rare earth lanthanides, are stable under a ground state, and emits fluorescentlight, being 540-600 nm in wavelength, under an excitation light source being 340-380 nm. The monoclonal antibody includes purified and mixed monoclonal antibodies which are sourced from a monoclonalantibody cell strain being direct at 2-6 difference cyclic citrullinated peptide antibodies and rheumatoid factor antigen epitopes. The kit has advantages of simple operation, rapid reaction, high sensitivity and strong specificity.

Description

Technical field: [0001] The invention relates to the technical field of fluorescent immunochromatography in medical immunology, in particular to a detection method for cyclic citrullinated peptide antibody and rheumatoid factor that can quickly and accurately perform quantitative analysis on cyclic citrullinated peptide antibody and rheumatoid factor Kit and preparation method thereof. Background technique: [0002] Rheumatoid arthritis (RA) is a multi-systemic inflammatory autoimmune disease mainly involving the peripheral joints. Rheumatic diseases characterized by variability of joint inflammation, joint destruction, and progressive sexual dysfunction have a prevalence of about 0.3%-0.6% in my country, and are one of the main causes of labor loss and disability in China. The disease is often misdiagnosed or cannot be diagnosed in time due to atypical symptoms in the early stage, and delayed treatment makes the prognosis of the disease poor. If the disease can be diagnosed...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/558G01N33/543G01N33/533
CPCG01N33/533G01N33/54313G01N33/558G01N33/577G01N33/6854G01N2800/102
Inventor 李红江王有志孙宁宁汪玉婷于鸿翔
Owner WEIHAI NEOPROBIO
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