Isolation and molecular characterization of an enteric isolate of the genotype-Ia bovine coronavirus with notable mutations in the receptor binding domain of the spike glycoprotein

Abid Ullah Shah; Phillip Gauger; Maged Gomaa Hemida

doi:10.1016/j.virol.2024.110313

Isolation and molecular characterization of an enteric isolate of the genotype-Ia bovine coronavirus with notable mutations in the receptor binding domain of the spike glycoprotein

Virology. 2024 Nov 29:603:110313. doi: 10.1016/j.virol.2024.110313. Online ahead of print.

Authors

Abid Ullah Shah¹, Phillip Gauger², Maged Gomaa Hemida³

Affiliations

¹ Department of Veterinary Biomedical Sciences, College of Veterinary Medicine, Long Island University, Brookville, NY, 11548NY, USA. Electronic address: abidullah.shah@liu.edu.
² Department of Veterinary Diagnostic and Production Animal Medicine, Veterinary Diagnostic University, Ames, IA, 50011, USA. Electronic address: pcgauger@iastate.edu.
³ Department of Veterinary Biomedical Sciences, College of Veterinary Medicine, Long Island University, Brookville, NY, 11548NY, USA. Electronic address: maged.hemida@liu.edu.

PMID: 39681059
DOI: 10.1016/j.virol.2024.110313

Abstract

BCoV new isolate was plaque purified, isolated, and propagated in vitro using MDBK and HRT-18. The full-length genome sequencing of this new BCoV isolate (31 Kbs) was drafted and deported in the GenBank. The genome organization is (5'-UTR-Gene-1-32kDa-HE-S-4.9 kDa-4.8 kDa-12.7 kDa-E-M-N-UTR-3'). Phylogenetic analysis based on the sequences of (the full-length genome, S, HE, and N) showed that the BCoV-13 clustered with other North American BCoV genotype I members. The sequence analysis shows several synonymous mutations among various domains of the S glycoprotein, especially the receptor binding domain. We found nine notable nucleotide deletions immediately downstream of the RNA binding domain of the nucleocapsid gene. Further gene function studies are encouraged to study the function of these mutations on the BCoV molecular pathogenesis and immune regulation. This research enhances our understanding of BCoV genomics and contributes to improved diagnostic and control measures for BCoV infections in cattle.

Keywords: BCoV; Enteric isolate; Genotype-1; HRT-18; IFA; Isolation; MDBK; NGS; Phylogenetic analysis; Plaque morphology; Plaque purification; RT-qPCR.