Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2012 May 4;156(3-4):418-24.
doi: 10.1016/j.vetmic.2011.11.013. Epub 2011 Nov 25.

Identification of a novel nidovirus associated with a neurological disease of the Australian brushtail possum (Trichosurus vulpecula)

M Dunowska  1 , P J Biggs, T Zheng, M R Perrott
Affiliations

Identification of a novel nidovirus associated with a neurological disease of the Australian brushtail possum (Trichosurus vulpecula)

M Dunowska et al. Vet Microbiol. .

Abstract

A novel, fatal neurological disease of the Australian brushtail possum (Trichosurus vulpecula) was first identified in 1995 in a research facility and subsequently in free-living possums in New Zealand and termed wobbly possum disease (WPD). The results of previous transmission studies suggested that the aetiological agent of WPD is most likely a virus. However, the identity of the presumed viral agent had not been elucidated. In the current report, we describe identification of a novel virus from tissues of WPD-affected possums using a combination of next generation sequencing and traditional molecular methods. The proportion of possums positive for the novel virus by PCR was significantly higher (p<0.0001) among animals with WPD than clinically healthy possums, strongly suggesting an aetiological involvement of the virus in WPD. Analysis of the partial genomic sequence of the putative WPD virus indicated that it is a novel nidovirus, most closely related to the current members of the family Arteriviridae.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
Sequencing strategy and organisation of the partial genome of the novel nidovirus. Top: Location of predicted ORFs (boundaries as per GenBank JN116253), ribosomal frameshift site (RSF), the initial NGS contig, overlapping amplicons (1–10), and 5′/3′ RACE products (primer sequences available on request). Middle: conserved protein domains including 3CLPRO (arterivirus NSP4 peptidase domain, NSP4_PRO), transmembrane (TM), RNA-dependent RNA polymerase (RdRp), putative zinc binding domain (ZBD), viral helicase 1 (HEL1), and Nidovirus EndoU-like (NendoU) domains. Bottom: Alignment of the putative ZBD of arteriviruses (abbreviations defined in Fig. 3) with the corresponding sequence of the putative wobbly possum disease virus (WPDV). Arrows point to conserved His/Cys residues (van Dinten et al., 2000) identified in the WPDV sequence; His/Cys residues identical in all five sequences are shaded.
Fig. 2
Fig. 2
An example of in situ hybridisation results: tissues from experimentally infected possums 674 (a and b) and 677 (c) with clinical wobbly possum disease (WPD) showing specific staining following hybridisation with the virus-specific probe (arrows in a); the same tissues hybridised with the control probe are shown in (d), (e) and (f).
Fig. 3
Fig. 3
Computer-based sequence analysis of the putative ORF8 protein. The same predicted features of the Equine arteritis virus (EAV, GenBank accession NC_002532.2) protein M are shown for comparison.
Fig. 4
Fig. 4
Maximum likelihood phylogenetic tree inferred from predicted amino acid sequence alignments of viral helicase (HEL1) rooted using Gill associated virus (GAV) as an outgroup. The numbers at the nodes depict % of bootstrap support. Accession numbers for the sequences are listed next to the virus names. Current taxonomic classification is included on the right. Abbreviations: wobbly possum disease virus (WPDV), White bream virus (WBV), Equine arteritis virus (EAV), Simian haemorrhagic fever virus (SHFV), Lactate dehydrogenase elevating virus (LDV), Porcine respiratory and reproductive syndrome virus (PRRSV), Avian infectious bronchitis virus (IBV), Transmissible gastroenteritis virus (TGE), Porcine epidemic diarrhoea virus (PEDV), Human coronavirus (HCoV), Severe acute respiratory syndrome (SARS), Murine hepatitis virus (MHV), and Porcine hemagglutinating encephalomyelitis virus (PHEV).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Brierley I., Jenner A.J., Inglis S.C. Mutational analysis of the “slippery-sequence” component of a coronavirus ribosomal frameshifting signal. J. Mol. Biol. 1992;227:463–479. - PMC - PubMed
    1. Carstens E.B. Ratification vote on taxonomic proposals to the International Committee on Taxonomy of Viruses (2009) Arch. Virol. 2010;155:133–146. - PMC - PubMed
    1. Cowan P.E. Brushtail possum. In: King C.M., editor. The Handbook of New Zealand Mammals. Oxford University Press; Melbourne: 2005.
    1. Cowley J.A., Dimmock C.M., Spann K.M., Walker P.J. Gill-associated virus of Penaeus monodon prawns: an invertebrate virus with ORF1a and ORF1b genes related to arteri- and coronaviruses. J. Gen. Virol. 2000;81:1473–1484. - PubMed
    1. de Vries A.A., Raamsman M.J., van Dijk H.A., Horzinek M.C., Rottier P.J. The small envelope glycoprotein (GS) of equine arteritis virus folds into three distinct monomers and a disulfide-linked dimer. J. Virol. 1995;69:3441–3448. - PMC - PubMed

Publication types

LinkOut - more resources