In situ hybridisation assay for localisation of rabbit calicivirus Australia-1 (RCV-A1) in European rabbit (Oryctolagus cuniculus) tissues

J Virol Methods. 2013 Mar;188(1-2):148-52. doi: 10.1016/j.jviromet.2012.11.043. Epub 2012 Dec 20.

Abstract

Recently, a new lagovirus enzootic in Australian wild rabbits was identified and described as rabbit calicivirus Australia-1 (RCV-A1). Unlike the closely related Rabbit Haemorrhagic Disease Virus (RHDV), which causes fulminant hepatitis and rabbit death, RCV-A1 does not appear to induce any clinical disease. RCV-A1 has been postulated to act as an imperfect natural vaccine to RHDV thus reducing RHDV-induced rabbit mortality, which is detrimental for bio-control of rabbits in Australia. This study was carried out to determine in which cells RCV-A1 replication occurs. An in situ hybridisation (ISH) protocol was developed using a RCV-A1 specific probe to localise the virus in rabbit tissues. The results were compared to those obtained with a quantitative RT-PCR assay that had previously been developed to measure RCV-A1 RNA in rabbit tissues. The histology of the tissues was also examined. ISH showed that virus replication, inferred by the presence of detectable RNA, was limited to a small number of epithelial cells towards the tip of the villi in the duodenum. Quantitative RT-PCR detected RCV-A1 RNA in jejunum, ileum and lymphoid tissue at day 3, 4 and 7 post-infection, but no hybridisation was detected in these tissues.

MeSH terms

  • Animals
  • Bunyaviridae Infections / veterinary*
  • Bunyaviridae Infections / virology
  • Histocytochemistry
  • Ileum / virology
  • In Situ Hybridization*
  • Jejunum / virology
  • Lagovirus / genetics
  • Lagovirus / isolation & purification*
  • Lymphoid Tissue / virology
  • Rabbits
  • Real-Time Polymerase Chain Reaction