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Improved antivirus strategies are needed to slow the spread of this pathogen.

Variant Rabbit Hemorrhagic Disease Virus in Young Rabbits, Spain. Investigation identified a rabbit hemorrhagic disease virus variant genetically related to apathogenic rabbit caliciviruses.

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RHDV is not cultivatable in cell culture; therefore, detection of virus genome, virions, and anti-RHDV antibodies and experimental infection of rabbits are required for diagnosis and virus characterization ( During September 2011–February 2012, our laboratory received liver samples from 9 rabbitries from 3 areas of northeastern Spain where acute outbreaks of RHD were occurring in adult rabbits and kits.

Effective inactivated vaccines prepared from liver extracts of rabbits experimentally infected with classic RHDV strains are used as a prophylactic and postoutbreak strategy to combat disease ().

The single serotype of RHDV is divided into 2 subtypes, classic RHDV and RHDVa.

), a virus with a positive-sense, single-stranded RNA genome of 7.4 kb.

Reverse transcription PCR was performed by using RNA extracted from 20 mg of liver samples using the mini RNAeasy RNA extraction kit (QIAGEN Iberia, Madrid, Spain), Superscript III reverse transcription (Invitrogen Corp., Carlsbad, CA, USA), LA-Taq polymerase (Takara Bio, Otsu, Japan), and forward and reverse primers annealing at nt 6056–60–6794, respectively (positions refer to the genomic sequence of RHDV Ast/89; Gen Bank accession no. A band of the expected size (738 bp) was purified after gel electrophoresis and sequenced; this isolate was named RHDV-N11 and deposited into Gen Bank (accession no. The sequenced region consisted of nt 6108–6716, corresponding to domains CDE and partial B and F domains of the VP1 capsid protein (Figure 1. Evolutionary relationships of rabbit hemorrhagic disease virus (RHDV) and related viruses.

A total of 38 nt sequences were analyzed: the isolate from this study, designated RHDV-N11 (Gen Bank accession no. Multiple sequence alignment and phylogenetic analysis were performed by using the RHDV-N11 VP1 sequence and 37 other sequences (18 classic RHDV, 12 RHDVa, and 6 RCV-like, with European brown hare syndrome virus as an outlier).

To confirm the presence of virions in the infected livers, liver homogenates (10% in sterile phosphate-buffered saline) were clarified by low-speed centrifugation followed by ultracentrifugation through a 30% sucrose cushion.

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