18D-10

Development of specific monoclonal antibody probes and user-friendly cost effective assay formats for the enumeration of Campylobacter jejuni in raw poultry is an important need for the poultry industries and for regulatory purposes. New rapid, portable and sensitive detection systems are also needed for improving the environmental monitoring for Campylobacter survival and persistence.

 

The objective was to develop a panel of new monoclonal antibodies exhibiting different reactive patterns and specificities to Campylobacter genus and C. jejuni cell surface antigens.

 

BALB/c female mice were immunized at four week intervals for a period of 12 months using a sonicated cell suspension of C. jejuni mix comprised of two human and two poultry isolates. Spleen cells from immunized BALB/c mice were fused with myeloma NS1 cells for the production of hybridoma clones. The hybridoma supernatants were screened by indirect ELISA against C. jejuni and C. coli whole cells to identify monoclonal antibody producing hybridoma clones.

 

We have developed a panel of new monoclonal antibodies (MAbs) exhibiting different reactivity patterns to Campylobacter and C. jejuni antigens. A total of 1518 hybridoma clones were produced, out of which 592 positive clones were identified by indirect ELISA against C. jejuni live spiral cells; based on this screening, 84 clones were preserved as frozen stocks. These MAbs were classified into two distinct groups based on current evaluation: (1) MAbs specific for C. jejuni without reacting with C. coli or other Campylobacter antigens; (2) MAbs reacting with C. jejuni, C. coli and other Campylobacter species.

 

The new MAbs will be extremely useful in leading to improved detection and enumeration strategies for C. jejuni and/or C. coli in raw foods and in environmental samples.