A rapid detection of Avian Pathogenic Escherichia coli (APEC) strains based on minimal number of virulence markers identified by whole genome sequencing.

Abstract

Background: Colibacillosis is an important epidemiological and economic issue in poultry farming and breeding. A common problem with avian pathogenic Escherichia coli strains (APEC) that cause this disease is a lack of an uniform identification system, resulting from a variety of serotypes, phylogenetic groups, sequence types and combinations of virulence factors. There are no clearly defined features that can be associated with pathogenicity. Therefore, without precise identification of pathogenic strains and differentiation from commensal strains, there is no possibility of appropriate selection of targeted therapy. The widespread use of whole genome sequencing (WGS) in recent years creates new possibilities in diagnostics. Therefore, the purpose of this study was to select features defining the APEC pathotype, based on next generation sequencing (NGS), and design a diagnostic test based on selected factors.

Results: A PCR diagnostic test is proposed. Three predictors of virulence were chosen according to in silico analysis: two virulence genes: iroC and hlyF, as well as one molecular marker of O78 serotype (wzx-O-antigen flippase of the O78 serotype). A choice of markers was supported by a chicken embryo model.

Conclusions: Whole genome sequencing of E. coli genomes allowed for the development of a rapid diagnostic method identifying pathogenic strains for poultry: APEC. The developed test can support field observations connected with the strain isolation source and clinical symptoms of the disease.