Isolation, propagation, genome analysis and epidemiology of HKU1 betacoronaviruses

Autor: Susmita Shrivastava, Amy Ransier, Andrew Berglund, Edison Luiz Durigon, Zhaohui Qian, Philip Weston, José Antonio Jerez, Christine C. Robinson, Luiz Gustavo Bentim Góes, Rebecca A. Halpin, Nadia Fedorova, Kathryn V. Holmes, Christopher D. Town, Samuel R. Dominguez
Rok vydání: 2014
Předmět:
Zdroj: Journal of General Virology
Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual)
Universidade de São Paulo (USP)
instacron:USP
ISSN: 1465-2099
0022-1317
DOI: 10.1099/vir.0.059832-0
Popis: From 1 January 2009 to 31 May 2013, 15 287 respiratory specimens submitted to the Clinical Virology Laboratory at the Children’s Hospital Colorado were tested for human coronavirus RNA by reverse transcription-PCR. Human coronaviruses HKU1, OC43, 229E and NL63 co-circulated during each of the respiratory seasons but with significant year-to-year variability, and cumulatively accounted for 7.4–15.6 % of all samples tested during the months of peak activity. A total of 79 (0.5 % prevalence) specimens were positive for human betacoronavirus HKU1 RNA. Genotypes HKU1 A and B were both isolated from clinical specimens and propagated on primary human tracheal–bronchial epithelial cells cultured at the air–liquid interface and were neutralizedin vitroby human intravenous immunoglobulin and by polyclonal rabbit antibodies to the spike glycoprotein of HKU1. Phylogenetic analysis of the deduced amino acid sequences of seven full-length genomes of Colorado HKU1 viruses and the spike glycoproteins from four additional HKU1 viruses from Colorado and three from Brazil demonstrated remarkable conservation of these sequences with genotypes circulating in Hong Kong and France. Within genotype A, all but one of the Colorado HKU1 sequences formed a unique subclade defined by three amino acid substitutions (W197F, F613Y and S752F) in the spike glycoprotein and exhibited a unique signature in the acidic tandem repeat in the N-terminal region of the nsp3 subdomain. Elucidating the function of and mechanisms responsible for the formation of these varying tandem repeats will increase our understanding of the replication process and pathogenicity of HKU1 and potentially of other coronaviruses.
Databáze: OpenAIRE