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Three-in-one time-resolved fluorescence immunochromatography kit and preparation method thereof

A fluorescence immunochromatography, time-resolved fluorescence technology, applied in analytical materials, biological material analysis, biological testing and other directions, can solve the problems of low specificity, misdiagnosis and missed diagnosis, low sensitivity, etc., to achieve good specificity, improve The effect of stability and high sensitivity

Pending Publication Date: 2022-03-04
武汉塞力斯生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because rheumatoid factor is interfered by certain antigens and antibody substances, false positives often occur and the specificity is not high. Often, it can only be diagnosed based on clinical symptoms and X-ray findings, resulting in misdiagnosis and missed diagnosis.
[0004] Secondly, the anti-cyclic citrullinated peptide antibody is an autoantibody with a synthetic cyclic citrullinated peptide as the antigen. It has a high specificity for the diagnosis of RA, but its sensitivity is low. There are still one-third of RA patients Plasma anti-cyclic citrullinated peptide antibodies were negative

Method used

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  • Three-in-one time-resolved fluorescence immunochromatography kit and preparation method thereof
  • Three-in-one time-resolved fluorescence immunochromatography kit and preparation method thereof
  • Three-in-one time-resolved fluorescence immunochromatography kit and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Step 1: Preparation of test strips

[0038] Preparation of conjugation pad 3 (anti-human IgG antibody immune complex labeled with time-resolved fluorescent microspheres)

[0039] 1) Preparation of time-resolved fluorescent solution microspheres: time-resolved fluorescent microspheres (purchased from Bangs Laboratories, Inc., with a diameter of 180 nm, doped with rare earth lanthanide elements, stable in the ground state, and emitted under the action of a 340 nm excitation light source Fluorescence at 600nm.) Diluted to 0.1% (W / W) with MES buffer solution, as fluorescent microsphere solution;

[0040]2) Microsphere activation: Dissolve EDC in MES buffer solution to 10 mg / ml, then quickly add it to the fluorescent microsphere solution, vortex and mix well, the amount of EDC solution added is 0.5 mg per milliliter of fluorescent microsphere solution, set Shake in a shaker at 37°C for 15 minutes.

[0041] 3) 15000rpm, 15min high-speed centrifugation, remove the supernatan...

Embodiment 2

[0067] Step 1: Preparation of test strips

[0068] Preparation of conjugation pad 3 (anti-human IgG antibody immune complex labeled with time-resolved fluorescent microspheres)

[0069] 1) Preparation of time-resolved fluorescent solution microspheres: Time-resolved fluorescent microspheres (purchased from Bangs Laboratories, Inc., with a diameter of 200 nm, doped with rare earth lanthanide elements, stable in the ground state, and emitted under the action of a 360 nm excitation light source Fluorescence at 620nm.) Diluted to 0.1% (W / W) with MES buffer solution, as fluorescent microsphere solution;

[0070] 2) Microsphere activation: Dissolve EDC in MES buffer solution to 10 mg / ml, then quickly add it to the fluorescent microsphere solution, vortex and mix well, the amount of EDC solution added is 0.5 mg per milliliter of fluorescent microsphere solution, set Shake in a shaker at 41°C for 18 minutes.

[0071] 3) Centrifuge at 15500 rpm for 18 minutes at high speed, remove th...

Embodiment 3

[0097] Step 1: Preparation of test strips

[0098] Preparation of conjugation pad 3 (anti-human IgG antibody immune complex labeled with time-resolved fluorescent microspheres)

[0099] 1) Preparation of time-resolved fluorescent solution microspheres: time-resolved fluorescent microspheres (purchased from Bangs Laboratories, Inc., with a diameter of 220 nm, doped with rare earth lanthanide elements, stable in the ground state, and emitted under the action of an excitation light source of 380 nm Fluorescence at 640nm.) Diluted to 0.1% (W / W) with MES buffer solution, as fluorescent microsphere solution;

[0100] 2) Microsphere activation: Dissolve EDC in MES buffer solution to 10 mg / ml, then quickly add it to the fluorescent microsphere solution, vortex and mix well, the amount of EDC solution added is 0.5 mg per milliliter of fluorescent microsphere solution, set Shake in a shaker at 45°C for 20min.

[0101] 3) 15000rpm, 20min high-speed centrifugation, remove the supernatan...

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Abstract

The invention discloses a three-in-one time-resolved fluorescence immunochromatography kit and a preparation method thereof. The three-in-one time-resolved fluorescence immunochromatography kit is composed of a PVC base plate, a glass fiber membrane, a conjugate pad, a nitrocellulose membrane and absorbent paper. A mouse anti-human IgG antibody marked by time-resolved fluorescent microspheres is contained in the combination pad; the nitrocellulose membrane comprises three detection lines: a T1 line is coated with cyclic citrullinated peptide recombinant protein; the T2 line is coated with n-pentamer protein 3 recombinant protein; the T3 line is coated with bispecific phosphatase 11 recombinant protein and a quality control C line; and the C line contains a goat anti-mouse IgG antibody. The kit can be used for rapidly and qualitatively detecting an anti-cyclic citrullinated peptide antibody or an anti-n-pentamer protein 3 antibody or an anti-bispecific phosphatase 11 antibody. The kit has the advantages of simplicity and convenience in operation, rapidness in reaction, high sensitivity and good specificity for early diagnosis of rheumatoid arthritis.

Description

technical field [0001] The invention relates to the technical field of medical kits, in particular to a three-in-one time-resolved fluorescence immunochromatography kit and a preparation method thereof. Background technique [0002] Rheumatoid arthritis (RA) is a common autoimmune disease characterized by chronic inflammation caused by synovial hyperplasia and cartilage destruction. This chronic inflammatory response in most cases of RA causes thickening of the synovial membrane and eventually destruction of the joint. In RA patients, about 70% of the symptoms of joint destruction progressively aggravate within 2 years. If the disease is not diagnosed and treated in time, erosive lesions of cartilage and bone will appear, resulting in damage to the joint structure, and eventually develop into joint deformity and ankylosis, resulting in There are varying degrees of disability, so early diagnosis is crucial. [0003] At present, the most commonly used clinical laboratory dia...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/564G01N33/558G01N33/58G01N33/548G01N33/552G01N33/533
CPCG01N33/564G01N33/558G01N33/582G01N33/587G01N33/548G01N33/552G01N33/533G01N2800/102
Inventor 姚松涛鲁翌李艳敏李靖雷之恒吴双
Owner 武汉塞力斯生物技术有限公司
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