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As the number of incidents of bacterial infections continues to rise around the globe, simpler, faster, and more sensitive diagnostic techniques are required to improve the safety of the food supply and to screen for potential bacterial infections in humans. We present here direct and indirect approaches for the detection of bacteria, which are based upon a combination of immunofluorescent staining and capillary electrophoresis. In the direct approach, Escherichia coli O157:H7 bacteria stained with fluorescein-tagged specific antibodies are detected by CE, while in the indirect approach fluorescein-tagged specific antibodies to E. coli are first captured by E. coli O157:H7 bacteria and then released and detected by CE. We have identified suitable bacteria staining and CE protocols, which involved a 10 mM Tris-borate-EDTA (TBE) buffer, 0.25 micro g antibody/1 million bacteria, and capillaries dynamically coated with poly-N-hydroxyethylacrylamide (polyDuramide). We have also successfully detected the presence of E. coli O157:H7 in contaminated meat. The total time required for analysis was 6-8 h, which is less than that realized in most commercial assays presently available.

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Membrane proteins of Pasteurella multocida have been shown previously to elicit protective immunity. We have identified an 87-kDa outer membrane antigen, Oma87, which is present in all 16 serotypes of P. multocida. The gene encoding this protein was cloned and sequenced and found to have significant similarity to the D15 protective surface antigen of Haemophilus influenzae. Oma87 was localized to the outer membrane of the cell, and proteinase K treatment suggested that the protein is surface exposed. Native and recombinant Oma87 were strongly immunostained by convalescent-phase antiserum, indicating that the protein is expressed in vivo. Specific Oma87 antiserum protected mice against homologous, lethal P. multocida challenge. These results suggest that Oma87 is a protective outer membrane antigen of P. multocida.