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Journal of Virology, January 2005, p. 884-895, Vol. 79, No. 2
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.2.884-895.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Characterization and Complete Genome Sequence of a Novel Coronavirus, Coronavirus HKU1, from Patients with Pneumonia

Patrick C. Y. Woo,1,2,{dagger} Susanna K. P. Lau,1,2,{dagger} Chung-ming Chu,3 Kwok-hung Chan,1 Hoi-wah Tsoi,1 Yi Huang,1 Beatrice H. L. Wong,1 Rosana W. S. Poon,1 James J. Cai,1 Wei-kwang Luk,4 Leo L. M. Poon,1,2 Samson S. Y. Wong,1,2 Yi Guan,1,2 J. S. Malik Peiris,1,2 and Kwok-yung Yuen1,2,{dagger}

Department of Microbiology,1 State Key Laboratory of Emerging Infectious Diseases, The University of Hong Kong,2 Division of Respiratory Medicine, Department of Medicine, United Christian Hospital,3 Department of Microbiology, Tseung Kwan O Hospital, Hong Kong4

Received 29 July 2004/ Accepted 3 September 2004

Despite extensive laboratory investigations in patients with respiratory tract infections, no microbiological cause can be identified in a significant proportion of patients. In the past 3 years, several novel respiratory viruses, including human metapneumovirus, severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV), and human coronavirus NL63, were discovered. Here we report the discovery of another novel coronavirus, coronavirus HKU1 (CoV-HKU1), from a 71-year-old man with pneumonia who had just returned from Shenzhen, China. Quantitative reverse transcription-PCR showed that the amount of CoV-HKU1 RNA was 8.5 to 9.6 x 106 copies per ml in his nasopharyngeal aspirates (NPAs) during the first week of the illness and dropped progressively to undetectable levels in subsequent weeks. He developed increasing serum levels of specific antibodies against the recombinant nucleocapsid protein of CoV-HKU1, with immunoglobulin M (IgM) titers of 1:20, 1:40, and 1:80 and IgG titers of <1:1,000, 1:2,000, and 1:8,000 in the first, second and fourth weeks of the illness, respectively. Isolation of the virus by using various cell lines, mixed neuron-glia culture, and intracerebral inoculation of suckling mice was unsuccessful. The complete genome sequence of CoV-HKU1 is a 29,926-nucleotide, polyadenylated RNA, with G+C content of 32%, the lowest among all known coronaviruses with available genome sequence. Phylogenetic analysis reveals that CoV-HKU1 is a new group 2 coronavirus. Screening of 400 NPAs, negative for SARS-CoV, from patients with respiratory illness during the SARS period identified the presence of CoV-HKU1 RNA in an additional specimen, with a viral load of 1.13 x 106 copies per ml, from a 35-year-old woman with pneumonia. Our data support the existence of a novel group 2 coronavirus associated with pneumonia in humans.

* Corresponding author. Mailing address: Department of Microbiology, The University of Hong Kong, University Pathology Building, Queen Mary Hospital, Hong Kong. Phone: (852) 28554892. Fax: (852) 28551241. E-mail: hkumicro{at}hkucc.hku.hk.

{dagger} P. C. Y. Woo, S. K. P. Lau, and K.-y. Yuen are all principal investigators and contributed equally to the manuscript.

Journal of Virology, January 2005, p. 884-895, Vol. 79, No. 2
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.2.884-895.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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