Autor(es): O'Brien Caitlin A, Hobson-Peters Jody, Yam Alice Wei Yee, Colmant Agathe M G, McLean Breeanna J, Prow Natalie A, Watterson Daniel, Hall-Mendelin Sonja, Warrilow David, Ng Mah-Lee, Khromykh Alexander A, Hall Roy A

Resumo: Mosquito-borne viruses encompass a range of virus families, comprising a number of significant human pathogens (e.g., dengue viruses, West Nile virus, Chikungunya virus). Virulent strains of these viruses are continually evolving - exp-ing their geographic range, thus rapid - sensitive screening assays are required to detect emerging viruses - monitor their prevalence - spread in mosquito populations. Double-str-ed RNA (dsRNA) is produced during the replication of many of these viruses as either an intermediate in RNA replication (e.g., flaviviruses, togaviruses) or the double-str-ed RNA genome (e.g., reoviruses). Detection - discovery of novel viruses from field - clinical samples usually relies on recognition of antigens or nucleotide sequences conserved within a virus genus or family. However, due to the wide antigenic - genetic variation within - between viral families, many novel or divergent species can be overlooked by these approaches. We have developed two monoclonal antibodies (mAbs) which show co-localised staining with proteins involved in viral RNA replication in immunofluorescence assay (IFA), suggesting specific reactivity to viral dsRNA. By assessing binding against a panel of synthetic dsRNA molecules, we have shown that these mAbs recognise dsRNA greater than 30 base pairs in length in a sequence-independent manner. IFA - enzyme-linked immunosorbent assay (ELISA) were employed to demonstrate detection of a panel of RNA viruses from several families, in a range of cell types. These mAbs, termed monoclonal antibodies to viral RNA intermediates in cells (MAVRIC), have now been incorporated into a high-throughput, economical ELISA-based screening system for the detection - discovery of viruses from mosquito populations. Our results have demonstrated that this simple system enables the efficient detection - isolation of a range of known - novel viruses in cells inoculated with field-caught mosquito samples, - represents a rapid, sequence-independent, - cost-effective approach to virus discovery.

Imprenta: PLoS Neglected Tropical Diseases, v. 9, n. 3, p. e0003629, 2015

Identificador do objeto digital: 10.1371/journal.pntd.0003629

Descritores: Chikungunya virus - Antigenic variation ; Chikungunya virus - Biosynthesis ; Chikungunya virus - Cell ; Chikungunya virus - Flaviviridae ; Chikungunya virus - Genome ; Chikungunya virus - Molecular structure ; Chikungunya virus - Pathogenesis ; Chikungunya virus - Proteins ; Chikungunya virus - RNA ; Chikungunya virus - Antibodies ; Chikungunya virus - Viral infections ; Chikungunya Virus - Virus ; Chikungunya virus - Molecular screening ; Chikungunya virus - Dengue ; Chikungunya virus - Immunology ; Chikungunya virus - Public health

Data de publicação: 2015

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