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

Method for detecting toxoplasma antibody

A technology of toxoplasma gondii and antibody, which is applied in the field of genetic engineering technology and diagnostic reagents, can solve the problems of false positives and low antigen specificity, and achieve the effects of less non-specificity, easy expression, purification and use, and simple process

Inactive Publication Date: 2010-10-20
CHANGZHOU 21ST CENTURY BIOTECH RES INSTCO
View PDF3 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problem that the existing ELISA detection antigen is not specific and is prone to false positives, the present invention discloses a method for detecting antibodies against Toxoplasma gondii by mixing recombinant proteins and polypeptides as antigens

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
  • Method for detecting toxoplasma antibody
  • Method for detecting toxoplasma antibody
  • Method for detecting toxoplasma antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Expression and purification of SAG1 recombinant protein and preparation of GRA6 polypeptide

[0022] Main reagents: His-TrapFastFlow column is produced by Phamachia Company, and other reagents are domestic or imported analytical reagents.

[0023] 1) Toxoplasma gondii SAG1 Screening of protein and GRA6 protein epitopes

[0024] Using ANTHEWIN and other software, the complete amino acid sequence of Toxoplasma gondii SAG1 protein (GeneBank, access number: S73634) and the entire amino acid sequence of GRA6 protein (GeneBank, access number: L33814) were analyzed by computer, and the antigenicity of SAG1 protein was screened out. The protein sequence of GRA6 is located from the 45th amino acid to the 198th amino acid; the strong antigenic epitope of GRA6 is located from the 55th to the 142nd amino acid. Their amino acid and DNA sequences are as follows:

[0025] The antigenic amino acid sequence (45-198aa) of the screened SAG1 protein is shown in SEQ NO.1; the ...

Embodiment 2

[0036] Example 2: Detection of SAG1 recombinant protein and GRA6 polypeptide

[0037] main reagent

[0038] 1) Anti-toxoplasma IgG serum was provided by Changzhou Maternal and Child Health Hospital.

[0039] 2) Enzyme-linked reaction materials: the enzyme-linked plate is a 96-well plate produced in Shenzhen, and the mouse anti-human (H+L) chain antibody labeled with horseradish peroxidase (HRP) was purchased from Jackson Company. Other materials are conventional materials for enzyme-linked reactions.

[0040] ELISA test: Indirect ELISA method is used to detect IgG antibody in serum.

[0041] The basic steps are:

[0042] The antigen was diluted to 100 μl with 50 mmol / L pH9.6 sodium carbonate, and then coated with enzyme-linked plates, 100 μl per well, overnight at 4°C. The next day, block with blocking solution (10 mmol / L phosphate buffer solution pH 7.4, containing 3% BSA), 100 μl per well, 37°C, 2h. After diluting the anti-toxoplasma IgG positive and negative serum to b...

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 relates to the field of a genetic engineering technology and a diagnostic reagent, in particular to a method for detecting a toxoplasma antibody by a toxoplasma recombinant protein and a synthetic peptide antigen mixed coating. In the method, a protein the sequence of which is SEQNO.1 and a polypeptide the sequence of which is SEQNO.3 are mixed to serve as an antigen to carry out ELISA (enzyme-linked immuno sorbent assay) detection. The invention maximumly shortens the protein length which is necessary for the antigen with better antigenicity, enhances the specificity of the detection and avoids the generation of false positive; the recombinant protein and the polypeptide are mixed to serve as the antigen, and thus, the sensitivity of the detection is improved, the preparation process is simplified to the maximum and the cost is saved.

Description

technical field [0001] The invention relates to the fields of genetic engineering technology and diagnostic reagents. Specifically, it relates to a method for detecting antibodies against toxoplasma gondii by mixing and coating toxoplasma recombinant protein and synthetic peptide antigen. Background technique [0002] Toxoplasmosis, also known as toxoplasmosis, is a zoonotic parasitic disease. The pathogen of toxoplasmosis is a coccidia. The whole life and development process of toxoplasma needs two hosts. Feline animals, such as domestic cats It is the final host, but its selection of intermediate hosts is extremely loose, and both mammals and birds can be used as intermediate hosts. If your pets (such as cats, dogs, birds, etc.) are infected with Toxoplasma gondii, or eat uncooked food infected with Toxoplasma gondii, you may get toxoplasmosis. About 1 / 3 of the world's people are infected Toxoplasma gondii. Normal people have no obvious clinical symptoms after being inf...

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
IPC IPC(8): G01N33/569
Inventor 曹利民杨永刚
Owner CHANGZHOU 21ST CENTURY BIOTECH RES INSTCO
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