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Kit for detecting anti-mutant citrulline waveform protein antibody and preparation method thereof

A technology for detecting reagents and vimentin, which is applied in the field of medicine and biological detection, and can solve the problems of poor diagnostic effect, insufficient sensitivity, insufficient specificity and sensitivity of markers, etc.

Pending Publication Date: 2020-06-23
SHENZHEN RUIMENG INNOVATION BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the diagnostic effect of early RA is not good, and most of the markers have insufficient specificity and sensitivity
The most widely used at present is anti-CCP, which is a kind of artificially synthesized polypeptide. Its most notable performance is that it has good specificity, but its sensitivity is not high enough (less than 70%)

Method used

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  • Kit for detecting anti-mutant citrulline waveform protein antibody and preparation method thereof
  • Kit for detecting anti-mutant citrulline waveform protein antibody and preparation method thereof
  • Kit for detecting anti-mutant citrulline waveform protein antibody and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0227] Example 1: Preparation of the first detection reagent

[0228] Process steps:

[0229] In 995g water, add 4.1g sodium dihydrogen phosphate, 5.64g disodium hydrogen phosphate dodecahydrate, 2g disodium edetate, 1g bovine serum albumin, 15g sodium chloride, 20g polyethylene glycol 8000, 2g Tween-20, 1g sodium azide, each time it is added, it needs to be stirred until it is completely dissolved, and the pH value is adjusted to 7.5; filter with a 0.22μm filter membrane to obtain 1L of the first detection reagent.

Embodiment 2

[0230] Example 2: Preparation of the second detection reagent

[0231] Raw material preparation:

[0232] The protein antigens of citrullinated vimentin and its mutants were prepared by the applicant.

[0233] The latex microspheres are polystyrene latex microspheres with carboxyl groups with a diameter of 200 nm, purchased from Japan JSR.

[0234] Process steps:

[0235] Prepare the second detection reagent as follows:

[0236] (1) Activation of carboxyl latex microspheres: Obtain 20g of 160nm carboxyl latex microspheres (100mg / mL), and add 10mL of 500mM Sulfo-NHS solution (pH 6.1) and 2mL of freshly prepared concentration of 20mg / mL Make up the EDAC solution with purified water to a final volume of 100mL, mix well, and mix for 30 minutes at room temperature (20℃-25℃);

[0237] (2) Coupling: Add the citrullinated vimentin and its mutant protein antigen to the system of step (1) according to the ratio of carboxyl latex microspheres: antigen mass ratio of 10:1, at room temperature, Mix f...

Embodiment 3

[0241] Example 3: Preparation of a kit for detecting anti-mutant citrulline vimentin antibodies including a first detection reagent and a second detection reagent

[0242] Process steps:

[0243] I. Preparation of the first detection reagent:

[0244] As shown in Example 1, the first detection reagent was prepared.

[0245] II. Preparation of the second detection reagent:

[0246] Prepare the second detection reagent as shown in Example 2.

[0247] III. Configuration kit:

[0248] The prepared first detection reagent and second detection reagent are configured into a kit according to a volume ratio of 4:1. For example, the packaging specification of the kit may be:

[0249] The first test reagent (4×40mL / bottle); and

[0250] The second detection reagent (2×20mL / bottle).

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Abstract

The invention belongs to the technical field of medicine and biochemistry, and particularly relates to an anti-mutant citrulline waveform protein antibody detection kit. The kit comprises a first detection reagent and a second detection reagent. The first detection reagent comprises a buffer agent, a stabilizer, an electrolyte, a sensitizer, a surfactant and a preservative. The second detection reagent comprises latex microspheres coupled with protein antigens of citrullinated waveform protein and mutants thereof, a buffer agent, a protein protective agent, an electrolyte and a preservative. The anti-mutant citrulline waveform protein antibody detection kit adopts a latex enhanced turbidimetric immunoassay method for determination, and has the advantages of rapidness, sensitivity, good accuracy, high specificity and the like compared with the traditional anti-mutant citrulline waveform protein antibody detection kit.

Description

Technical field [0001] This application relates to the field of medicine and biological detection. Specifically, this application relates to a kit for detecting anti-mutant citrulline vimentin antibodies and a preparation method thereof. Background technique [0002] Rheumatoid arthritis (RA) is a chronic, systemic disease with inflammatory synovitis with unknown etiology. Chronic inflammation of the synovial membrane can cause the destruction of articular cartilage, hard bones and soft tissue around the joint, and the hyperplasia or fibrosis of the synovial membrane. Rheumatoid arthritis is manifested in the limitation of multiple joints such as fingers, wrists, knees, etc., causing muscle atrophy, joint swelling and deformation, and ultimately resulting in loss of function. Rheumatoid arthritis is common among young and middle-aged people, especially women. It is characterized by symmetrical damage to most joints. The lesions start from the distal small joints with swelling an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/543G01N33/6893G01N2800/102
Inventor 李昀轩林潮喜
Owner SHENZHEN RUIMENG INNOVATION BIOTECHNOLOGY CO LTD
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