European and America type pig breeding and syndrome virus antibody identification and diagnosis reagent kit preparation method

A syndrome virus and differential diagnosis technology, which is applied in the field of preparation of European and American type porcine reproduction and syndrome virus antibody differential diagnosis kits, and can solve the problems of large differences between types

Inactive Publication Date: 2008-08-06
INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
View PDF1 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The virus structural proteins encoded by ORF7 of both have good antigenicity, and the nucleotide sequence of N protein is conserved within the type, and the difference between the types is large

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
  • European and America type pig breeding and syndrome virus antibody identification and diagnosis reagent kit preparation method
  • European and America type pig breeding and syndrome virus antibody identification and diagnosis reagent kit preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0022] 1) Expression and purification of target protein:

[0023] A prokaryotic expression vector pGEX-6P-1- N; Design and synthesize the nucleic acid sequences for European PRRSV N protein epitopes 2aa~12aa, 40aa~46aa and 67aa~128aa, establish a prokaryotic recombinant expression vector PQE30-N for European PRRSV N protein, and use purification kits protein purification.

[0024] After renaturation, the recombinant protein GST-N was eluted with reduced glutathione to obtain the target protein (as shown in Figure 1). After affinity chromatography purification, the purity of the target protein can reach more than 90%. After the recombinant fusion protein HIS-N is purified by nickel affinity chromatography column, there is a very small amount of impurity protein, and its purity can reach more than 95%, and the purification effect reaches the expected purpose (as shown in Figure 2).

[0025] 1.1 Western-Blot analysis of recombinant protein

[0026] Western-Blot analysis was p...

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 provides a preparation method of a European American type pig propagation and syndrome virus antibody differential diagnosis Kit. The method constructs the pronucleus expression vector pGEX-6P-1-N aimed at American type PRRSV N protein regarding the gene deficiency of the conserved epitopes 25aa-30aa, 37aa-52aa and 50aa-66aa of European and American type strain in an ORF7 gene of a PRRSV VR-2332 strain. Regarding the amino acid sequence of the European type PRRSV N protein epitopes 2aa-12aa, 40aa-46aa and 67aa-128aa, the method establishes the pronucleus recombination expression vector PQE30-N of the European type PRRSV N protein. The recombination plasmid expresses successfully in the bacillus coli, and an indirect ELISA diagnosis Kit for PRRS antibody detection is prepared through using the expression depurated North American type N protein. The coincidence rates of the antibody Kit and an IDEXX Kit both reach 90 percent, and the depurated European type protein serves as enveloping antigen, thereby optimizing the European type PRRSV antibody detection indirection ELISA method, and laying a foundation for PRRS antibody typing detection.

Description

1. Technical field [0001] The invention relates to veterinary disease diagnosis technology in the field of biotechnology, in particular to a preparation method of a European and American type porcine reproduction and syndrome virus antibody differential diagnosis kit. 2. Background technology [0002] The occurrence and prevalence of PRRS have caused considerable economic losses to the pig industry in my country. But at present, the prevention and control of the disease is still a difficult problem faced by countries all over the world. In order to better prevent and control the occurrence and prevalence of this disease, it is urgent to establish a reliable method suitable for large-scale diagnosis and detection of PRRS. Common diagnostic methods for PRRS infection include cell culture and RT-PCR. However, these diagnostic methods are time-consuming, heavy in workload, and require special instruments and equipment, which are far from meeting the current needs of diagnosing...

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 Applications(China)
IPC IPC(8): G01N33/569
Inventor 王金宝张祯涛李俊任慧英吴家强张秀美丛晓燕温建新周顺
Owner INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY 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