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Int. J. Mol. Sci. 2016, 17(5), 691; doi:10.3390/ijms17050691

Isolation and Characterization of Dromedary Camel Coronavirus UAE-HKU23 from Dromedaries of the Middle East: Minimal Serological Cross-Reactivity between MERS Coronavirus and Dromedary Camel Coronavirus UAE-HKU23

1
State Key Laboratory of Emerging Infectious Diseases, the University of Hong Kong, Pokfulam, Hong Kong
2
Department of Microbiology, the University of Hong Kong, Pokfulam, Hong Kong
3
Research Centre of Infection and Immunology, the University of Hong Kong, Pokfulam, Hong Kong
4
Carol Yu Centre for Infection, the University of Hong Kong, Pokfulam, Hong Kong
5
Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Zhejiang University, Hangzhou 310006, China
6
Central Veterinary Research Laboratory, Dubai, UAE
These authors contributed equally to this work.
*
Authors to whom correspondence should be addressed.
Academic Editor: Charles A. Collyer
Received: 17 March 2016 / Revised: 21 April 2016 / Accepted: 25 April 2016 / Published: 7 May 2016
(This article belongs to the Special Issue Host-Microbe Interaction)
View Full-Text   |   Download PDF [3577 KB, uploaded 7 May 2016]   |  

Abstract

Recently, we reported the discovery of a dromedary camel coronavirus UAE-HKU23 (DcCoV UAE-HKU23) from dromedaries in the Middle East. In this study, DcCoV UAE-HKU23 was successfully isolated in two of the 14 dromedary fecal samples using HRT-18G cells, with cytopathic effects observed five days after inoculation. Northern blot analysis revealed at least seven distinct RNA species, corresponding to predicted subgenomic mRNAs and confirming the core sequence of transcription regulatory sequence motifs as 5′-UCUAAAC-3′ as we predicted previously. Antibodies against DcCoV UAE-HKU23 were detected in 58 (98.3%) and 59 (100%) of the 59 dromedary sera by immunofluorescence and neutralization antibody tests, respectively. There was significant correlation between the antibody titers determined by immunofluorescence and neutralization assays (Pearson coefficient = 0.525, p < 0.0001). Immunization of mice using recombinant N proteins of DcCoV UAE-HKU23 and Middle East respiratory syndrome coronavirus (MERS-CoV), respectively, and heat-inactivated DcCoV UAE-HKU23 showed minimal cross-antigenicity between DcCoV UAE-HKU23 and MERS-CoV by Western blot and neutralization antibody assays. Codon usage and genetic distance analysis of RdRp, S and N genes showed that the 14 strains of DcCoV UAE-HKU23 formed a distinct cluster, separated from those of other closely related members of Betacoronavirus 1, including alpaca CoV, confirming that DcCoV UAE-HKU23 is a novel member of Betacoronavirus 1. View Full-Text
Keywords: coronavirus; dromedary camel; isolation; characterization coronavirus; dromedary camel; isolation; characterization
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Woo, P.C.Y.; Lau, S.K.P.; Fan, R.Y.Y.; Lau, C.C.Y.; Wong, E.Y.M.; Joseph, S.; Tsang, A.K.L.; Wernery, R.; Yip, C.C.Y.; Tsang, C.-C.; Wernery, U.; Yuen, K.-Y. Isolation and Characterization of Dromedary Camel Coronavirus UAE-HKU23 from Dromedaries of the Middle East: Minimal Serological Cross-Reactivity between MERS Coronavirus and Dromedary Camel Coronavirus UAE-HKU23. Int. J. Mol. Sci. 2016, 17, 691.

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