Porcine circovirus disease detection method based on ORF2 (Open Reading Frame 2) antigen domain

A technology of GST-ORF2-E and functional area, which is applied in botany equipment and methods, biochemical equipment and methods, viral peptides, etc., can solve the problems of time-consuming and labor-intensive, virus-dispersing infected cells, etc., to improve sensitivity and specificity , the effect of improving specificity

Inactive Publication Date: 2011-01-05
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
View PDF0 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the experimental operation of these methods is time-consuming and laborious, and there is a risk of spreading the virus ...

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
  • Porcine circovirus disease detection method based on ORF2 (Open Reading Frame 2) antigen domain
  • Porcine circovirus disease detection method based on ORF2 (Open Reading Frame 2) antigen domain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0029] 1. Primer design and PCR amplification of target gene

[0030] Data report (MahéD, Blanchard P, Truong C, Arnauld C, Le Cann P, CarioletR, Madec F, Albina E, Jestin A. 2000. Differential recognition of ORF2protein from type 1 and type 2porcine circoviruses and identification of immunorelevant epitopes. J Gen Virol. 81 (Pt 7): 1815-1824.), there are five main immune response regions on the protein encoded by PCV2, one of which is on the ORF1 protein coding region, and the other four on ORF2. Because only the antigen functional area (113-147) on ORF2 is the most effective, it may become the main antigen for the development of ELISA kits for detecting PCV2 infection in herds. In the present invention, the amino acid segment (113-147) on ORF2 is selected as the target antigen for cloning and expression, and its gene fragment is 102 bp in length. Using the whole PCV2 ORF2 gene as a template, two specific primers were designed, and the gene fragments located in the 113-147aa sp...

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 a PCV (Porcine Circovirus) disease detection method based on an ORF2 (Open Reading Frame 2) antigen domain. The method comprises the following steps of: obtaining GST-ORF2-E infusion protein through the fusion expression of a special amino acid segment (113-147aa) on PCV2 ORF2 as a target with GST (Glutathione S-Transferase); and purifying the fusion protein by utilizing affinity chromatography and using the purified fusion protein as the antigen to coat an ELISA (Enzyme-Linked Immunosorbent Assay) 96 pore plate and establishing an ELISA method. The special amino acid segment selected by the method is a segment with higher antigenicity on the PCV2 ORF2 and used as the coated antigen in ELISA to improve the sensitivity and the specificity of a traditional PCV2 ELISA method. Compared with a commonly applied immunofluorescence detection method, the specificity and the sensitivity of the method are respectively 87.7 percent and 93.57 percent. The ORF2 without antigen cross between the PCV1 and the PCV 2 can be used for differentiating and judging the infection of the PCV1 and the PCV 2. The method can be used for differentiating animals infected by the PCV1 and the PCV 2.

Description

Technical field [0001] The present invention relates to animal husbandry and veterinary science and technology, in particular to a method for detecting porcine circumcision disease based on the ORF2 (based on open reading frame 2) antigen functional region. Background technique [0002] Porcine Circumcision Disease II (PCV2) is the main pathogen that causes multiple system failure syndrome (PMWS) in weaned piglets. The disease mainly causes weaned piglets to develop stunted growth, progressive weight loss, pale skin or jaundice, difficulty breathing, diarrhea, etc. Symptoms, clinically due to secondary bacterial and viral infections, the clinical symptoms of the disease are complex and diverse. Therefore, after pigs are infected with PCV-2, PMWS can occur at 5-12 weeks of age, with an morbidity rate of 5%-30% and a mortality rate of 10%-30%. The disease has been reported in many countries and regions in the world, and the virus is also widespread in our country's herds. The dis...

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): G01N33/569C12N15/34C12N15/70C07K14/01
Inventor 郭慧琛孙世琪刘湘涛尹双辉尚佑军殷宏才学鹏
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