Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Myositis and myasthenia gravis autoantibody detection kit and method in human body fluid

A myasthenia gravis and detection kit technology, which is applied in the field of biomedicine, can solve the problems of time-consuming detection process, large demand for antibodies, and low detection sensitivity, and achieve the effects of short detection cycle, reduced pollution, and simple operation

Active Publication Date: 2022-03-04
SHAANXI MYBIOTECH CO LTD
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problems existing in the prior art, the purpose of the present invention is to provide a detection kit and method for autoantibodies in myositis and myasthenia gravis in human body fluid, so as to solve the problem of high detection cost and antibody demand of existing detection methods or detection kits. Large size, low detection sensitivity, time-consuming detection process and other problems

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Myositis and myasthenia gravis autoantibody detection kit and method in human body fluid
  • Myositis and myasthenia gravis autoantibody detection kit and method in human body fluid
  • Myositis and myasthenia gravis autoantibody detection kit and method in human body fluid

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0092] The target antigen Ag in the detection kit of the present invention can be obtained through the existing technology, and can also be obtained through the following methods of the present invention, preferably the preparation method of the present invention. Specifically, the preparation method of myositis and myasthenia gravis autoantibody detection material in human body fluid of the present invention comprises the following steps:

[0093] Step 1, obtain the CDS sequence of the target antigen Ag as the target gene, and insert the target gene with restriction sites into the pET28a plasmid vector to obtain the recombinant plasmid vector pET28a-Ag; Ag represents myositis and myasthenia gravis autoantibody corresponding One of the target antigens;

[0094] Step 1.1, obtaining the CDS sequence of the autoantibody target antigen Ag by artificial synthesis or PCR method as the target gene, and adding NheI / NotI restriction sites at both ends of the target gene;

[0095] Step...

Embodiment 1

[0128]This example discloses a method for preparing myositis and myasthenia gravis autoantibody detection materials in human body fluid. The target antigen in this example is the target antigen MDA5 of myositis disease, and the negative control antigen is GAPDH. The preparation method is specific include:

[0129] 1. Plasmid construction:

[0130] Obtain the CDS sequences of autoantibody target antigens MDA5 and GAPDH by artificial synthesis or PCR method as the target gene, and add NheI / NotI restriction sites at both ends of the target gene;

[0131] Insert the target gene with a restriction site into the pET28a vector, the insertion site is NheI / NotI, to obtain a recombinant vector, which is named pET28a-MDA5 and pET28a-GAPDH, specifically:

[0132] Take the glycerol strain of the pET28a plasmid and inoculate it into 3 mL LB liquid medium resistant to kanamycin, place it in a constant temperature incubator at 37°C, and shake the bacteria overnight at 220r / min. On the secon...

Embodiment 2

[0151] This embodiment discloses a detection kit, which includes a test strip, a working solution and a blocking protein solution;

[0152] The test strip is the test strip corresponding to the target antigen in Example 1. Specifically, the test strip includes a backing plate, a carrier film with a control line and a detection line, an absorbent pad, a sample pad and a gold standard pad; the carrier film detects The target antigen Ag is immobilized on the point, and Ag represents a certain antigen in the target antigen corresponding to the autoantibody of myositis and myasthenia gravis disease; the negative control antigen is immobilized on the control line. Water-absorbing pads, carrier film, sample pads, and gold standard pads are all laid on the backing plate. figure 1 , figure 2 shown.

[0153] Wherein, the working solution is a mixed solution of 1×PBS, 0.5% Triton X-100, and 0.04% EDTA, and the pH of the working solution is 6.8.

[0154] The blocking protein solution ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a kit and a method for detecting autoantibodies of myositis and myasthenia gravis in human body fluid. The detection kit includes a test strip, a working solution and a closed protein solution; the detection method includes mixing the working solution and the closed protein solution to form sample dilution solution, add the sample to be tested in the sample diluent; add the sample to be tested dropwise on the sample pad of the test strip; remove the absorbent pad and sample pad on the test strip, wash the carrier film, and then wash the carrier film The absorbent pad and the gold standard pad are fixed on both ends of the carrier film; the end of the gold standard pad of the test strip is immersed in the working solution, and the color development is observed. In the detection process of the present invention, the sample chromatographic method is inverted chromatography, and the carrier membrane washing method is ultrasonic washing, which effectively increases the detection sensitivity and specificity, and the strong positive serum can still detect antibodies after being diluted more than 10,000 times.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a detection kit and method for myositis and myasthenia gravis autoantibodies in human body fluid. Background technique [0002] Idiopathic inflammatory myopathies (IIMs) are rare autoimmune diseases characterized by muscle inflammation leading to proximal muscle atrophy and disability, various skin rashes, ulcers, malignancies, etc. symptom. Autoantibodies can be detected in body fluids of more than 50% of IIM patients, so autoimmunity is considered to play an important role in the pathogenesis of myositis. Current studies have shown that these autoantibodies target nuclear and cytoplasmic proteins, and they are usually divided into two subgroups, myositis-specific autoantibodies (MSAs) and myositis-associated autoantibodies (Myositis-associated autoantibodies, MAAs). A large number of clinically relevant studies have shown that MSAs and MAAs are important mark...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/564G01N33/558C07K14/47C12N15/70
CPCG01N33/564G01N33/558C07K14/47C12N15/70C07K1/22C07K1/18C07K1/20G01N2800/2878G01N2800/10
Inventor 闫亚平李怡婷冯昆封雪裴元元李科
Owner SHAANXI MYBIOTECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com