scholarly journals Recovery of Infectious Rabbit Hemorrhagic Disease Virus from Rabbits after Direct Inoculation with In Vitro-Transcribed RNA

2006 ◽  
Vol 80 (13) ◽  
pp. 6597-6602 ◽  
Author(s):  
Guangqing Liu ◽  
Yuying Zhang ◽  
Zheng Ni ◽  
Tao Yun ◽  
Zutian Sheng ◽  
...  
Keyword(s):  
Disease Virus ◽  
Infectious Clone ◽  
Full Length ◽  
Rabbit Hemorrhagic Disease Virus ◽  
Progeny Virus ◽  
Cdna Clones ◽  
Hemorrhagic Disease ◽  
Rabbit Hemorrhagic Disease ◽  
Rna Transcripts

ABSTRACT We report the first full-length infectious clone of strain JX/CHA/97 of rabbit hemorrhagic disease virus (RHDV). The transcripts from the full-length cDNA clones were infectious when they were directly injected into rabbits. The sequence of the virus recovered from the rabbits was identical to that of the injected RNA transcripts. The cDNA clone was engineered to contain one silent nucleotide change to create an EcoRV site (A to T at nucleotide 2908). The genetic marker was retained in the recovered progeny virus. The transfection of RNA transcripts into RK-13 cells resulted in the synthesis of viral antigens, indicating that the cDNA clones were replication competent. This stable infectious molecular clone should be an important tool for developing a better understanding of the molecular biology and pathogenesis of RHDV.

scholarly journals A Review on the Methods Used for the Detection and Diagnosis of Rabbit Hemorrhagic Disease Virus (RHDV)

2021 ◽  
Vol 9 (5) ◽  
pp. 972
Author(s):  
Joana Abrantes ◽  
Ana M. Lopes
Keyword(s):  
Disease Virus ◽  
Diagnostic Tests ◽  
High Throughput Sequencing ◽  
Current Knowledge ◽  
European Rabbit ◽  
Rabbit Hemorrhagic Disease Virus ◽  
Hemorrhagic Disease ◽  
Rabbit Hemorrhagic Disease ◽  
Detection And Diagnosis

Since the early 1980s, the European rabbit (Oryctolagus cuniculus) has been threatened by the rabbit hemorrhagic disease (RHD). The disease is caused by a lagovirus of the family Caliciviridae, the rabbit hemorrhagic disease virus (RHDV). The need for detection, identification and further characterization of RHDV led to the development of several diagnostic tests. Owing to the lack of an appropriate cell culture system for in vitro propagation of the virus, much of the methods involved in these tests contributed to our current knowledge on RHD and RHDV and to the development of vaccines to contain the disease. Here, we provide a comprehensive review of the RHDV diagnostic tests used since the first RHD outbreak and that include molecular, histological and serological techniques, ranging from simpler tests initially used, such as the hemagglutination test, to the more recent and sophisticated high-throughput sequencing, along with an overview of their potential and their limitations.

scholarly journals Recombinant VP60 in the form of virion-like particles as a potential vaccine against rabbit hemorrhagic disease virus.

2006 ◽  
Vol 53 (2) ◽  
pp. 371-376 ◽  
Author(s):  
Beata Gromadzka ◽  
Bogusław Szewczyk ◽  
Grazyna Konopa ◽  
Andrzej Fitzner ◽  
Andrzej Kesy
Keyword(s):  
Disease Virus ◽  
Rna Virus ◽  
Full Length ◽  
Rabbit Hemorrhagic Disease Virus ◽  
Hemorrhagic Disease ◽  
Rabbit Hemorrhagic Disease ◽  
Vp60 Gene ◽  
A Genome ◽  
Potential Vaccine ◽  
Domestic Rabbits

Rabbit hemorrhagic disease virus (RHDV) which causes a highly contagious disease of wild and domestic rabbits belongs to the family Caliciviridae. It is a small, positive single-stranded RNA virus with a genome of 7.5 kb and has a diameter of approximately 40 nm. In negatively stained electron micrographs the virus shows typical calicivirus morphology with regularly arranged cup-shaped structures on the surface. It is a major pathogen of rabbits in many countries. Vp60 - a coat protein of molecular mass around 60 kDa is the major antigen of RHDV. It is present as 90 dimeric units per virion particle. We have expressed VP60 gene in the baculovirus system with the aim to use it as a potential vaccine against RHDV and a diagnostic reagent in immunological tests. cDNA of the vp60 gene of strain SGM, was cloned into a baculovirus transfer vector as full-length gene, as well as truncated gene lacking 600 5'-terminal nucleotides. The sequence of SGM VP60 differed markedly from that of the reference strain. Full-length recombinant VP60 protein from the SGM strain self-assembled to form virus-like particles (VLPs). These particles observed by electron microscopy were morphologically similar to native virions and were able to agglutinate human group 0 erythrocytes. After immunization the recombinant particles induced RHDV-specific antibodies in rabbits and guinea pigs. Rabbits immunized with the VLPs were fully protected against challenge with a virulent RHDV.

scholarly journals Expression of Enzymatically Active Rabbit Hemorrhagic Disease Virus RNA-Dependent RNA Polymerase in Escherichia coli

1998 ◽  
Vol 72 (4) ◽  
pp. 2999-3004 ◽  
Author(s):  
Ana López Vázquez ◽  
José M. Martín Alonso ◽  
Rosa Casais ◽  
José A. Boga ◽  
Francisco Parra
Keyword(s):  
Escherichia Coli ◽  
Rna Polymerase ◽  
Disease Virus ◽  
Rabbit Hemorrhagic Disease Virus ◽  
Hemorrhagic Disease ◽  
Coding Region ◽  
Recombinant Polypeptide ◽  
Rna Dependent Rna Polymerase ◽  
Rabbit Hemorrhagic Disease

ABSTRACT The rabbit hemorrhagic disease virus (RHDV) (isolate AST/89) RNA-dependent RNA-polymerase (3Dpol) coding region was expressed in Escherichia coli by using a glutathioneS-transferase-based vector, which allowed milligram purification of a homogeneous enzyme with an expected molecular mass of about 58 kDa. The recombinant polypeptide exhibited rifampin- and actinomycin D-resistant, poly(A)-dependent poly(U) polymerase. The enzyme also showed RNA polymerase activity in in vitro reactions with synthetic RHDV subgenomic RNA in the presence or absence of an oligo(U) primer. Template-size products were synthesized in the oligo(U)-primed reactions, whereas in the absence of added primer, RNA products up to twice the length of the template were made. The double-length RNA products were double stranded and hybridized to both positive- and negative-sense probes.

scholarly journals ATP Binding and ATPase Activities Associated with Recombinant Rabbit Hemorrhagic Disease Virus 2C-Like Polypeptide

2000 ◽  
Vol 74 (22) ◽  
pp. 10846-10851 ◽  
Author(s):  
M. Soledad Marín ◽  
Rosa Casais ◽  
José M. Martín Alonso ◽  
Francisco Parra
Keyword(s):  
Disease Virus ◽  
Site Directed Mutagenesis ◽  
Rabbit Hemorrhagic Disease Virus ◽  
Atp Binding ◽  
Hemorrhagic Disease ◽  
Rabbit Hemorrhagic Disease ◽  
Carboxy Terminal Region ◽  
Carboxy Terminal

ABSTRACT The carboxy-terminal region of the rabbit hemorrhagic disease virus p37 polyprotein cleavage product has been expressed inEscherichia coli as a glutathione S-transferase (GST) fusion protein. The recombinant GST-Δ2C protein showed in vitro ATP-binding and ATPase activities. Site-directed mutagenesis studies of the conserved residues G522 and T529 in motif A, D566 and E567 in motif B, and K600 in motif C were also performed. These results provide the first experimental characterization of a 2C-like ATPase activity in a member of the Caliciviridae.

scholarly journals Detection of Viral Proteins after Infection of Cultured Hepatocytes with Rabbit Hemorrhagic Disease Virus

1998 ◽  
Vol 72 (5) ◽  
pp. 4492-4497 ◽  
Author(s):  
Matthias König ◽  
Heinz-Jürgen Thiel ◽  
Gregor Meyers
Keyword(s):  
Disease Virus ◽  
Genome Organization ◽  
Gradient Centrifugation ◽  
Liver Perfusion ◽  
Viral Proteins ◽  
Rabbit Hemorrhagic Disease Virus ◽  
Hemorrhagic Disease ◽  
Rabbit Hemorrhagic Disease ◽  
Infected Cells

ABSTRACT The calicivirus rabbit hemorrhagic disease virus (RHDV), which replicates predominantly in the livers of infected rabbits, cannot be propagated in tissue culture. To enable the performance of in vitro studies, rabbit hepatocytes were isolated by liver perfusion and gradient centrifugation. After inoculation with purified RHDV, more than 50% of the cells proved to be infected. Protein analyses led to the detection of 13 RHDV-specific polypeptides within the infected cells. These proteins were assigned to defined regions of the viral genome, resulting in a refined model of RHDV genome organization.

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