Haemophilus parasuis positive indirect hemagglutination diagnostic reagent containing Haemophilus parasuis oppa recombinant protein antigen and preparation method thereof

A technology of Haemophilus suis disease and Haemophilus suis, which is applied in the field of molecular biology, can solve the problems that a single antibody detection reagent cannot meet the production needs, the composition of the whole bacteria ultra-breaking antigen is complex, and lack of specificity, etc., to achieve strong promotion and Application value, intuitive detection results, and strong specificity

Active Publication Date: 2016-03-02
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these diagnostic methods have certain limitations. The antigenic composition of the whole bacteria is complex and lacks specificity. Although the capsular polysaccharide antigen has good specificity, it can only detect a single antibody of the corresponding serotype.
However, in China, the prevalent serotypes of Haemophilus parasuis are relatively complex, mainly including serotypes 4, 5, 12, and 13. Other serotypes can occasionally occur clinically, so a single antibody detection reagent cannot meet the actual conditions. Production needs

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
  • Haemophilus parasuis positive indirect hemagglutination diagnostic reagent containing Haemophilus parasuis oppa recombinant protein antigen and preparation method thereof
  • Haemophilus parasuis positive indirect hemagglutination diagnostic reagent containing Haemophilus parasuis oppa recombinant protein antigen and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] 1. Haemophilus parasuis OppA Acquisition of the target gene

[0021] 1.1 Reaction system

[0022] Premixed Enzyme (Bao Bio): 12.5 , Primers (Bao Biology): 1 each , template 1 , Sterile double water: 9.5 ;

[0023] Primer: Upstream primer 5'-ATGCAAACAACCTTTACC-3'

[0024] Downstream primer 5'-TTACTGCTTAATGATATA-3';

[0025] 1.2 Amplification conditions

[0026] 94°C, 5min; 94°C, 50s; 51°C, 40s; 72°C, 100s; 35 cycles.

[0027] 2. Acquisition of OppA target protein

[0028] 2.1 will amplify OppA The target gene is connected with the cloning vector pMD19-T and transformed into Escherichia coli JM109; the positive monoclonal strain is picked and the plasmid is extracted; endonuclease EcoR I and xho I carried out double digestion of the plasmid and Pet-30a, then connected it with T4 ligase, and then transformed it into the expression strain BL21 (DE3), and induced expression with IPTG at a final concentration of 0.08mmol / L, such as figure 1 A schematic ...

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 fast, convenient and practical Haemophilus parasuis disease positive indirect hemagglutination diagnostic reagent containing Haemophilus parasuis OppA recombinant protein antigen and a preparation method thereof, and provides a parasuis Haemophilus OppA recombinant protein antigen and its coding gene. The beneficial effects of the present invention are: 1. the detection process is simple, fast, accurate, practical, and has strong promotion and application value; 2. high sensitivity, strong specificity and good stability; 3. the diagnostic reagent can detect side effects Antibody of Haemophilus suis serotype 15; ④ safe and simple, no complicated testing instruments and equipment are needed, and the testing results are intuitive.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to Haemophilus parasuis OppA recombinant protein antigen and its coding gene, and a positive indirect hemagglutination diagnostic reagent for Haemophilus parasuis containing the Haemophilus parasuis OppA recombinant protein antigen. Background technique [0002] Haemophilus parasuis disease is caused by Haemophilus parasuis ( Haemophilus Parasuis , Hps) causes an infectious disease in pigs characterized by fibrinous serositis, polyarthritis and meningitis. The disease occurs all over the world, usually seen in pigs aged 5-8 weeks, the incidence rate is generally 10%-15%, and the mortality rate can reach 50% in severe cases. In recent years, the infection rate and the resulting mortality of this disease in large-scale pig farms have been increasing day by day, and it has become one of the more serious bacterial diseases that endanger the pig industry. [0003] Haemophilus parasuis ...

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): C07K14/285C12N15/31C12N15/63C12N5/10C12N1/21C12N15/70G01N33/68C12R1/21
Inventor 逯忠新高鹏程贺英储岳峰赵萍
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap