Tibet Orbivirus, a novel Orbivirus species isolated from Anopheles maculatus mosquitoes in Tibet, China

PLoS One. 2014 Feb 12;9(2):e88738. doi: 10.1371/journal.pone.0088738. eCollection 2014.

Abstract

Background: The genus Orbivirus includes a number of important pathogenic viruses, including Bluetongue virus (BTV), African horse sickness virus (AHSV), and Epizootic hemorrhagic disease virus (EHDV). In this study we describe the isolation and characterization of an Orbivirus strain isolated from Anopheles maculatus mosquitoes collected in Tibet, China.

Methods and results: Initial viral screening identified a viral strain (XZ0906) that caused significant cytopathic effect (CPE) in BHK-21 cells, including rounding, cell rupture, and floating. Although CPE was not observed in insect cells (C6/36), these cells supported viral replication. Polyacrylamide gel analysis revealed a genome consisting of 10 segments of double-stranded RNA (dsRNA), with a distribution pattern of 3-3-3-1. 454 high throughput sequencing of culture supernatant was used for viral identification. Complete genome sequencing was performed by Sanger sequencing in combination with 5'-RACE and 3'-RACE. Sequence analysis demonstrated that all 5'- and 3'- untranslated regions (UTRs) for each of the 10 genome segments contained a series of six highly conserved nucleotides. In addition, homology analysis and phylogenetic analysis based on amino acid sequence was completed, and all results show that virus XZ0906 was not a member of any known species or serotype of Orbivirus, indicating it to be a new species within the genus Orbivirus.

Conclusions: The isolated Orbivirus strain was designated Tibet Orbivirus, TIBOV to denote the location from which it was isolated. TIBOV is a novel orbivirus species which is isolated from Anopheles maculatus mosquitoes collected in Tibet, China.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions / genetics
  • 5' Untranslated Regions / genetics
  • Amino Acid Sequence
  • Animals
  • Anopheles / virology*
  • Base Sequence
  • Cell Line
  • Cricetinae
  • Genome, Viral / genetics
  • Orbivirus / genetics
  • Orbivirus / isolation & purification*
  • Orbivirus / physiology
  • Sequence Analysis, DNA
  • Sequence Analysis, RNA
  • Tibet
  • Viral Proteins / chemistry
  • Viral Proteins / genetics

Substances

  • 3' Untranslated Regions
  • 5' Untranslated Regions
  • Viral Proteins

Grants and funding

This work is supported by grants from National Natural Science Foundation of China (81290342), The Ministry of Science and Technology, China (2011CB504702) and Development Grant of State Key Laboratory for Infectious Disease Prevention and Control (2008SKLID105). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.