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

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...

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 during the preparation of infected cells, especially these diagnostic methods require experienced researchers to make correct judgments

Images