Detection of live and heat‐treatedSalmonella enteritidisby a D1‐serospecific anti‐lipopolysaccharide O‐9 monoclonal antibody

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Abstract
A murine monoclonal antibody 2F11 (IgG2a) against Salmonella enteritidis was produced by a fusion of P3X63 Ag8.653 myeloma cells with splenocytes of a mouse immunized with heat‐attenuated (80°C, 20 min) S. enteritidis cells. The specificity of this antibody was tested in an indirect ELISA and sodium dodecyl sulphate polyacrylamide gel electrophoresis followed by immunoblotting. The monoclonal antibody was specific to D1‐serogroup Salmonella, exhibiting the highest reactivity with all tested phage types of S. enteritidis (1, 4, 8, 13, and 13a). The monoclonal antibody was reactive with heat‐attenuated, as well as live S. enteritidis cells. In addition, this antibody exhibited high and equal avidity to lipopolysaccharides isolated from S. enteritidis, regardless of phage types. The monoclonal antibody 2F11 proved to be specific to lipopolysaccharide O‐9 present in D1‐serogroup Salmonella. Immunoblotting and ELISA results demonstrated that the epitope recognized by this antibody was partially composed of tyvelose and mannose. It was also determined by the nature of glycosidic bonds between monosaccharides in the polysaccharide backbone region. Employing poly‐L‐lysine precoated microplates, this antibody exhibited the detection limit of 104 S. enteritidis cells ml‐1 of buffer, as assessed by ELISA.

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