Isolation and purification of O and H antigens from Salmonella Enteritidis as diagnostic tool

Lipopolysaccharide (LPS) and flagellin are the major antigenic structures of Salmonella Enteritidis. Purified antigens are essential components for the diagnosis of and research on Salmonella Enteritidis. In this study, LPS were isolated and purified with modified hot phenol–water procedure in a yield of 6%. The flagellin isolation and purification were done by strong shaking with glass pearl beads with 3.9 mg/ml protein content. Purity was demonstrated on SDS-PAGE, ELISA and with colorimetric assay for both isolated antigens. The antigen-mediated ELISA results exhibited immunoreactive properties for both LPS and flagellar antigen specific for Salmonella Enteritidis O:9 and H:g,m, respectively. The findings demonstrated low reactivity with Mab O:9 for isolated LPS due to their mono-epitopic specificity. However, isolated flagellin reactivity was high with Mab H:g,m related to their cross-reactivity potential between H:g and H:m. Additionally in this study. the results indicated that similar antigenic structures have alternative potential for antigenic isolation of Salmonella Enteritidis, such as Salmonella Gallinarum for LPS antigen and Salmonella Adeyo for flagellar antigen isolation.