Isolation And Identification Of Etiological Agent And The Study Of Indirect ELISA For Exudative Epidermitis

Staphylococcus hyicus is the causal agent of the Exudative Epidermitis(EE) seen in piglets. The classic disease occurs as an acute or peracute infection in suckling and newly weaned piglets in which a generalized epidermitis may lead to dehydration and death. Piglets usually develop the disease between 4-6days and 5-6 weeks of age. Clinical signs begin with dejection and a reddish or coppery skin color. Thin, pale brown scales of exudates develop in the axillae and groin and within 3-5 days spread to all parts of the body and rapidly become dark in color and greasy in texture. Anorexia and dehydration are features of this disease.Until now, there were only two aspects been reported. One was about isolated and identification of Staphylococcus hyicus, the other was the treatment by antibiotic of Exudative Epidermitis. There was no studied on Etiology,Pathogenesis and Diagnosis.Those detection was time consuming and laborious. It could quickly detect a lot of pathological samples by Indirect ELISE with epidemiology investigation. The ExhB gene of Staphylococcus hyicus was amplified by PCR . The products were cloned into a prokaryotic expression vector pET32a. After certification by restriction analysis, PCR amplification and sequence analysis, the recombinant expression plasmid pET32a-ExhB was transformed into E.coli BL21(DE3) and induced by IPTG.. After soluble analysis the fusion protein was furified. The culture was collected and tested by SDS-PAGE, Western blot and indirect ELISA. The result shows in comparison with GENE BANK data the homologies of the nueleofide sequence is 98%. The 30ku fusion protein was existed in the form of inclusion body and can be recognized by the positive serum of Staphylococcus hyicus. Indirect ELISA of Exudative epidermitis had been established, which do good for the study of the kit.