title: In situ hybridisation assay for localisation of rabbit calicivirus Australia-1 (RCV-A1) in European rabbit (Oryctolagus cuniculus) tissues.
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.
authors.affiliation: Commonwealth Scientific and Industrial Research Organisation, Division of Ecosystem Sciences, Australia.
authors.forename: Marion
authors.initials: M
authors.lastname: Hoehn
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Kerr
authors.forename: Tanja
authors.initials: T
authors.lastname: Strive
mesh_terms: D000818:Animals; D002044:Bunyaviridae Infections / Q000662:veterinary* / Q000821:virology; D006651:Histocytochemistry; D007082:Ileum / Q000821:virology; D017403:In Situ Hybridization; D007583:Jejunum / Q000821:virology; D029321:Lagovirus / Q000235:genetics / Q000302:isolation & purification*; D008221:Lymphoid Tissue / Q000821:virology; D011817:Rabbits; D060888:Real-Time Polymerase Chain Reaction