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Avian pathogenic  Escherichia coli  (APEC) is the bacterial pathogen of poultry colibacillosis, which causes significant economic losses to the poultry industry. The lack of an effective vaccine against multiple serotypes and the emergence of multi-resistant isolates have made the control of avian colibacillosis troublesome. To identify conserved potential vaccine candidates, 58 genomes of APEC were obtained (54 sequenced by our laboratory and four downloaded from NCBI). A reverse vaccinology (RV) method based on the pangenome – called Pan-RV analysis – was performed in APEC-protective protein mining for the first time. Finally, four proteins were selected, and their immunoreactivity with anti-O1, O2, and O78 serum was verified by western blotting. Our  in silico  method of analysis will pave the way for rapid screening of vaccine candidates and will lay the foundation for the development of a highly effective subunit vaccine controlling APEC infection. RESEARCH HIGHLIGHTSPan-RV analysis was used for the first time in the discovery of APEC-protective proteins.A total of 53 potential protective proteins were screened out.Four proteins were verified as potential vaccine candidates using western blotting. Pan-RV analysis was used for the first time in the discovery of APEC-protective proteins. A total of 53 potential protective proteins were screened out. Four proteins were verified as potential vaccine candidates using western blotting.

Combining pangenome analysis to identify potential cross-protective antigens against avian pathogenic Escherichia coli