Isolation and genetic characterization of MERS-CoV from dromedary camels in the United Arab Emirates

Abdelmalik Ibrahim Khalafalla; Hassan Zackaria Ali Ishag; Hamdoon Ismail Abdulla Albalushi; Zulaikha Mohamed Abdel-Hameed Al-Hammadi; Saeed Mohamed Saeed Al Yammahi; Asma Abdi Mohamed Shah; Salama Suhail Mohammed Al Muhairi

doi:10.3389/fvets.2023.1182165

Isolation and genetic characterization of MERS-CoV from dromedary camels in the United Arab Emirates

Front Vet Sci. 2023 Aug 31:10:1182165. doi: 10.3389/fvets.2023.1182165. eCollection 2023.

Authors

Abdelmalik Ibrahim Khalafalla¹, Hassan Zackaria Ali Ishag¹, Hamdoon Ismail Abdulla Albalushi¹, Zulaikha Mohamed Abdel-Hameed Al-Hammadi¹, Saeed Mohamed Saeed Al Yammahi¹, Asma Abdi Mohamed Shah¹, Salama Suhail Mohammed Al Muhairi¹

Affiliation

¹ Biosecurity Affairs Division, Development and Innovation Sector, Abu Dhabi Agriculture and Food Safety Authority (ADAFSA), Abu Dhabi, United Arab Emirates.

Abstract

Background: The study of coronaviruses has grown significantly in recent years.Middle East respiratory syndrome coronavirus (MERS-CoV) replicates in various cell types, and quick development has been made of assays for its growth and quantification. However, only a few viral isolates are now available for investigation with full characterization. The current study aimed to isolate MERS-CoV from nasal swabs of dromedary camels and molecularly analyze the virus in order to detect strain-specific mutations and ascertain lineage classification.

Methods: We isolated the virus in Vero cells and adapted it for in vitro cultivation. The isolates were subjected to complete genome sequencing using next-generation sequencing followed by phylogenetic, mutation, and recombination analysis of the sequences.

Results: A total of five viral isolates were obtained in Vero cells and adapted to in vitro cultures. Phylogenetic analysis classified all the isolates within clade B3. Four isolates clustered close to the MERS-CoV isolate camel/KFU-HKU-I/2017 (GenBank ID: MN758606.1) with nucleotide identity 99.90-99.91%. The later isolate clustered close to the MERS-CoV isolate Al-Hasa-SA2407/2016 (GenBank ID: MN654975.1) with a sequence identity of 99.86%. Furthermore, the isolates contained several amino acids substitutions in ORF1a (32), ORF1ab (25), S (2), ORF3 (4), ORF4b (4), M (3), ORF8b (1), and the N protein (1). The analysis further identified a recombination event in one of the reported sequences (OQ423284/MERS-CoV/dromedary/UAE-Al Ain/13/2016).

Conclusion: Data presented in this study indicated the need for continuous identification and characterization of MERS-CoV to monitor virus circulation in the region, which is necessary to develop effective control measures. The mutations described in this investigation might not accurately represent the virus's natural evolution as artificial mutations may develop during cell culture passage. The isolated MERS-CoV strains would be helpful in new live attenuated vaccine development and efficacy studies.

Keywords: MERS-CoV; camel; characterization; coronavirus; dromedary; isolation; zoonotic disease.