Isolation, propagation, genome analysis and epidemiology of HKU1 betacoronaviruses

摘要: From 1 January 2009 to 31 May 2013, 15 287 respiratory specimens submitted the Clinical Virology Laboratory at 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 seasons but with significant year-to-year variability, cumulatively accounted 7.4–15.6 % all samples months peak activity. A total 79 (0.5 % prevalence) positive betacoronavirus HKU1 RNA. Genotypes B both isolated from clinical propagated on primary tracheal–bronchial epithelial cells cultured air–liquid interface neutralized in vitro intravenous immunoglobulin polyclonal rabbit antibodies spike glycoprotein HKU1. Phylogenetic analysis deduced amino acid sequences seven full-length genomes viruses glycoproteins four additional three Brazil demonstrated remarkable conservation these genotypes circulating Hong Kong France. Within genotype A, one formed a unique subclade defined substitutions (W197F, F613Y S752F) exhibited signature acidic tandem repeat N-terminal region nsp3 subdomain. Elucidating function mechanisms responsible formation varying repeats will increase our understanding replication process pathogenicity potentially other coronaviruses.