Viral RNA intermediates as targets for detection and discovery of novel and emerging mosquito-borne viruses

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. and Hall, Roy A. (2015) Viral RNA intermediates as targets for detection and discovery of novel and emerging mosquito-borne viruses. PLoS Neglected Tropical Diseases, 9 3: 1-27. doi:10.1371/journal.pntd.0003629

Author 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.
Title Viral RNA intermediates as targets for detection and discovery of novel and emerging mosquito-borne viruses
Journal name PLoS Neglected Tropical Diseases   Check publisher's open access policy
ISSN 1935-2735
Publication date 2015-03-23
Year available 2015
Sub-type Article (original research)
DOI 10.1371/journal.pntd.0003629
Open Access Status DOI
Volume 9
Issue 3
Start page 1
End page 27
Total pages 27
Place of publication San Francisco, CA United States
Publisher Public Library of Science
Language eng
Subject 2739 Public Health, Environmental and Occupational Health
2725 Infectious Diseases
Abstract 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 and expanding their geographic range, thus rapid and sensitive screening assays are required to detect emerging viruses and monitor their prevalence and spread in mosquito populations. Double-stranded 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-stranded RNA genome (e.g., reoviruses). Detection and discovery of novel viruses from field and clinical samples usually relies on recognition of antigens or nucleotide sequences conserved within a virus genus or family. However, due to the wide antigenic and genetic variation within and 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 and 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 and discovery of viruses from mosquito populations. Our results have demonstrated that this simple system enables the efficient detection and isolation of a range of known and novel viruses in cells inoculated with field-caught mosquito samples, and represents a rapid, sequence-independent, and cost-effective approach to virus discovery.
Keyword Infectious Diseases
Tropical Medicine
Infectious Diseases
Tropical Medicine
Q-Index Code C1
Q-Index Status Confirmed Code
Grant ID DP120103994 12/14
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
School of Chemistry and Molecular Biosciences
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Citation counts: TR Web of Science Citation Count  Cited 19 times in Thomson Reuters Web of Science Article | Citations
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Created: Sat, 09 May 2015, 00:07:15 EST by Mrs Louise Nimwegen on behalf of School of Chemistry & Molecular Biosciences