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Archerfield, Australia

Jansen C.C.,CSIRO | Jansen C.C.,University of Queensland | Jansen C.C.,Metro North Public Health Unit | Hemmerter S.,University of Technology, Sydney | And 4 more authors.
Australian Journal of Entomology | Year: 2013

Members of the Culex sitiens subgroup of mosquitoes are important arbovirus vectors in the Australasian region. However, some species in this group, particularly Cx.annulirostris and Cx.palpalis, are difficult to distinguish based on morphological characters alone. We evaluated the reliability of morphological characters commonly used in taxonomic keys against a PCR-based diagnostic tool that examined the ribosomal ITS1 gene region. Although reliable morphological characters allow identification, molecular identification remains the most accurate means for distinguishing Cx. annulirostris and Cx. palpalis. © 2013 Australian Entomological Society.

O'Brien C.A.,University of Queensland | Hobson-Peters J.,University of Queensland | Yam A.W.Y.,University of Queensland | Yam A.W.Y.,Cancer Science Institute | And 9 more authors.
PLoS Neglected Tropical Diseases | Year: 2015

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. © 2015 O’Brien et al.

Huang B.,Public Health Virology Laboratory | Allcock R.,University of Western Australia | Allcock R.,Medical Center | Warrilow D.,Public Health Virology Laboratory
Virology Reports | Year: 2016

Arboviruses circulate in the biota of northern Australia, sometimes causing disease in animals and humans. Five different arboviruses, isolated and stored for over 30 years, were re-cultured and sequenced using high-throughput sequencing technology (Ion Torrent PGM) for identification and genetic characterization. Phylogenetic analysis, primarily of sequence fragments of the large segment, indicated classification as members of the Family Bunyaviridae (Belmont, Little Sussex, Parker's Farm, and Thimiri viruses). Another was identified as a member of the Family Rhabdoviridae (Beatrice Hill virus) based on phylogenetic analysis of the RNA polymerase region. © 2016 Published by Elsevier B.V.

Warrilow D.,Public Health Virology Laboratory | Watterson D.,University of Queensland | Hall R.A.,University of Queensland | Davis S.S.,Australian Department of Primary Industries and Fisheries | And 8 more authors.
PLoS ONE | Year: 2014

Here we describe Casuarina virus (CASV), a new virus in the family Mesoniviridae. This is the first report of a mesonivirus in Australia, which extends the geographical range of this virus family to 3 continents. The virus was isolated in 2010 from Coquillettidia xanthogaster mosquitoes during surveillance in the suburbs of Darwin, the capital of the Northern Territory. Cryo-electron microscopy of the CASV virions revealed spherical particles of 65 nm in size with large club-shaped projections of approximately 15 nm in length. The new virus was most closely related to Alphamesonivirus 1, the only currently recognized species in the family. In 2013 a further 5 putative new mesonivirus species were described: Hana, Méno, Nsé, Moumo and Dak Nong viruses. The evolutionary distance between CASV and two of its closest relatives, Cavally and Hana viruses (Jones-Taylor-Thornton distance of 0.151 and 0.224, respectively), along with its isolation from a different genus of mosquitoes captured on a separate continent indicate that CASV is a new species. © 2014 Warrilow et al.

Shinwari M.W.,Biosecurity science Laboratory | Annand E.J.,Randwick Equine Center | Driver L.,Biosecurity science Laboratory | Warrilow D.,Public Health Virology Laboratory | And 8 more authors.
Veterinary Microbiology | Year: 2014

In May 2013, the first cases of Australian bat lyssavirus infections in domestic animals were identified in Australia. Two horses (filly-H1 and gelding-H2) were infected with the Yellow-bellied sheathtail bat (YBST) variant of Australian bat lyssavirus (ABLV). The horses presented with neurological signs, pyrexia and progressing ataxia. Intra-cytoplasmic inclusion bodies (Negri bodies) were detected in some Purkinje neurons in haematoxylin and eosin (H&E) stained sections from the brain of one of the two infected horses (H2) by histological examination. A morphological diagnosis of sub-acute moderate non-suppurative, predominantly angiocentric, meningo-encephalomyelitis of viral aetiology was made. The presumptive diagnosis of ABLV infection was confirmed by the positive testing of the affected brain tissue from (H2) in a range of laboratory tests including fluorescent antibody test (FAT) and real-time PCR targeting the nucleocapsid (N) gene. Retrospective testing of the oral swab from (H1) in the real-time PCR also returned a positive result. The FAT and immunohistochemistry (IHC) revealed an abundance of ABLV antigen throughout the examined brain sections. ABLV was isolated from the brain (H2) and oral swab/saliva (H1) in the neuroblastoma cell line (MNA). Alignment of the genome sequence revealed a 97.7% identity with the YBST ABLV strain. © 2014.

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