Full-length genome reveals genetic diversity and extensive recombination patterns of Saudi GI-1 and GI-23 genotypes of infectious bronchitis virus

Background: Despite numerous genetic studies on Infectious Bronchitis Virus (IBV), many strains from the Middle East remain misclassified or unclassified. Genotype 1 (GI-1) is found globally, while genotype 23 (GI-23) has emerged as the predominant genotype in the Middle East region, evolving continuously through inter- and intra-genotypic recombination. The GI-23 genotype is now enzootic in Europe and Asia.

Methods: Over a 24-month period from May 2022 to June 2024, 360 samples were collected from 19 layer and 3 broiler poultry farms in central Saudi Arabia. The chickens exhibited reduced laying rates and symptoms such as weakness and respiratory distress, while broilers showed respiratory issues. Samples, including tracheal swabs and various tissue specimens, were pooled, homogenized, and stored at -20 °C prior to PCR analysis. The samples underwent virus isolation in embryonated chicken eggs, RNA extraction using automated systems, and detection of IBV through real-time RT-PCR targeting a conserved 5'-UTR fragment. Full-length genome sequencing was performed, and recombination analysis was conducted using RDP 4.6.

Results: Saudi IBV strains were found to cluster into genotypes GI-1 and GI-23.1. The study identified critical amino acid substitutions in the hypervariable regions of the spike protein and detected recombination events in the ORF1ab, N, M, 3ab, and 5ab genes, with nsp3 of the ORF1ab showing the greatest number of recombination events.

Conclusion: The multiple inter- and intra-genotypic recombination events that were detected in different genes indicate that the circulating IBV strains do not share a single ancestor but have emerged through successive recombination events.