Isolation and Characterization of a Novel Betacoronavirus Subgroup A ...

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Coronavirus, Rabbit Coronavirus HKU14, from Domestic Rabbits. Susanna K. P. ..... TTCTGAGGTGTCTTCAG-3= (for human coronavirus OC43 [HCoV. OC43]) ...

Isolation and Characterization of a Novel Betacoronavirus Subgroup A Coronavirus, Rabbit Coronavirus HKU14, from Domestic Rabbits Susanna K. P. Lau,a,b,c,d Patrick C. Y. Woo,a,b,c,d Cyril C. Y. Yip,d Rachel Y. Y. Fan,d Yi Huang,d Ming Wang,e Rongtong Guo,e Carol S. F. Lam,d Alan K. L. Tsang,d Kenneth K. Y. Lai,d Kwok-Hung Chan,d Xiao-Yan Che,f Bo-Jian Zheng,a,b,c,d and Kwok-Yung Yuena,b,c,d State Key Laboratory of Emerging Infectious Diseases,a Research Centre of Infection and Immunology,b Carol Yu Centre for Infection,c and Department of Microbiology,d The University of Hong Kong, Hong Kong, and Guangzhou Center for Disease Control and Prevention,e and Center of Laboratory, Zhujiang Hospital, Southern Medical University,f Guangzhou, China

We describe the isolation and characterization of a novel Betacoronavirus subgroup A coronavirus, rabbit coronavirus HKU14 (RbCoV HKU14), from domestic rabbits. The virus was detected in 11 (8.1%) of 136 rabbit fecal samples by reverse transcriptase PCR (RT-PCR), with a viral load of up to 108 copies/ml. RbCoV HKU14 was able to replicate in HRT-18G and RK13 cells with cytopathic effects. Northern blotting confirmed the production of subgenomic mRNAs coding for the HE, S, NS5a, E, M, and N proteins. Subgenomic mRNA analysis revealed a transcription regulatory sequence, 5=-UCUAAAC-3=. Phylogenetic analysis showed that RbCoV HKU14 formed a distinct branch among Betacoronavirus subgroup A coronaviruses, being most closely related to but separate from the species Betacoronavirus 1. A comparison of the conserved replicase domains showed that RbCoV HKU14 possessed

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