Heterologous expression of the membrane protein OmpA from bovine Pasteurella multocida and establishment of an indirect ELISA for antibody detection
Year: 2026
Type: article
Abstract: Reliable serological tools are needed for surveillance of Pasteurella multocida antibodies. Four P. multocida membrane proteins were expressed in a prokaryotic system and screened by Western blotting and indirect ELISA. Among them, a dilution-refolded OmpA preparation retained serum immunoreactivity and met the antigen-selection criterion under the tested conditions. The optimized assay used a nominal total-protein coating concentration of 1 µg/mL, 1% skim milk blocking at 37 °C for 1 h, serum diluted 1:400, secondary antibody diluted 1:25,000, and 15 min colour development. In the preliminary analytical-specificity panel, the available PmB-positive serum and positive sera against eight other bovine respiratory pathogens remained below the lower decision threshold. PmA-positive serum remained test-positive at a dilution of 1:1280, and the intra-assay and inter-assay coefficients of variation were below 5%. Within the vaccination-defined 60-serum development panel, ROC analysis yielded an AUC of 0.969 and apparent sensitivity and specificity of 95.00% each relative to the predefined serum groups. These estimates are preliminary because the panel was derived from a single ranch, the same 20 predefined negative-reference sera were also used to establish the decision thresholds, and no independent reference method was applied.
Source: Scientific Reports
Institution: Inner Mongolia University of Technology
Language: English
Subfield: Microbiology
Field: Immunology and Microbiology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,690
OpenAlex ID: W7223031389