Conventional and real time RT-PCR assays for the detection and differentiation of variant rabbit hemorrhagic disease virus (RHDVb) and its recombinants

Dalton, Kevin P.; Arnal, José Luis; Benito, Alfredo A.; Chacón, G.; Alonso, J.M.; Parra, Francisco

doi:10.1016/j.jviromet.2017.10.009

Cited by 25 publications

(42 citation statements)

References 27 publications

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“…Rabbit hemorrhagic disease (RHD) is a highly contagious disease characterized by acute liver damage and disseminated intravascular coagulation (DIC) (Alonso et al, 1998; Jung et al, 2000). The first outbreak of RHD was reported in 1984 in the Jiangsu Province of China (Zhu et al, 2016) and has quickly spread to most parts of the world (Calvete et al, 2018; Dalton et al, 2018). This has led to the deaths of millions of rabbits and hares, representing a serious threat to their populations.…”

Section: Introductionmentioning

confidence: 99%

Rabbit Hemorrhagic Disease Virus Non-structural Protein 6 Induces Apoptosis in Rabbit Kidney Cells

Chen

Liu

et al. 2019

Front. Microbiol.

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Rabbit hemorrhagic disease (RHD) is a highly contagious disease caused by rabbit hemorrhagic disease virus (RHDV). Previous research has shown that RHDV induces apoptosis in numerous cell types, although the molecular mechanisms underlying the apoptosis induced by RHDV are not well understood. One possible factor is non-structural protein 6 (NSP6), a 3C-like protease that plays an important role in processing viral polyprotein precursors into mature non-structural proteins. To fully establish a role for NSP6, the present study examined the effects of ectopic expression of the protein in rabbit (RK13) and human (HeLa and HepG2) cells. We found that NSP6 suppressed cell viability and promoted apoptosis in all three cell types in a dose-dependent manner. We also identified increased caspase-3, -8, and -9 activities in RK13 cell, and an increased Bax to Bcl2 mRNA ratio. Mechanistically, the ability of NSP6 to induce apoptosis was impaired by mutation of the catalytic His27 residue. Our study has shown that RHDV NSP6 can induce apoptosis in host cells and is likely an important contributor to RHDV-induced apoptosis and pathogenesis.

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Section: Introductionmentioning

confidence: 99%

Rabbit Hemorrhagic Disease Virus Non-structural Protein 6 Induces Apoptosis in Rabbit Kidney Cells

Chen

Liu

et al. 2019

Front. Microbiol.

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“…Another approach is the use of an endogenous amplification control consisting of the beta-actin gene, which has been incorporated into the RT-rtPCR designed for RHDVb detection. 22 The endogenous amplification control confirmed the correct extraction and amplification performance of the assay.…”

Section: Detection Of Lagomorph Calicivirusesmentioning

confidence: 67%

“…71 The emergence of new RHDV-2 strains in rabbits initiated research toward the development of strain-specific TaqMan RT-rtPCR methods. 16,22,30 The primers and probes targeted a highly conserved fragment of the RHDV-2 VP60 gene, with only a few nucleotide mismatches compared to RHDV genes as well as to non-and low-pathogenic RCVs. Although these methods are able to detect RHDV-2 strains, full sequence analysis of the complete VP60 gene is necessary when differentiation between vaccine and field virus strains is needed.…”

Section: Detection Of Lagomorph Calicivirusesmentioning

confidence: 99%

Molecular methods in detection and epidemiologic studies of rabbit and hare viruses: a review

Kwit

Rzeżutka

2019

J VET Diagn Invest

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Various PCR-based assays for rabbit viruses have gradually replaced traditional virologic assays, such as virus isolation, because they offer high-throughput analysis, better test sensitivity and specificity, and allow vaccine and wild-type virus strains to be fully typed and differentiated. In addition, PCR is irreplaceable in the detection of uncultivable or fastidious rabbit pathogens or those occurring in low quantity in a tested sample. We provide herein an overview of the current state of the art in the molecular detection of lagomorph viral pathogens along with details of their targeted gene or nucleic acid sequence and recommendations for their application. Apart from the nucleic acids-based methods used for identification and comprehensive typing of rabbit viruses, novel methods such as microarray, next-generation sequencing, and mass spectrometry (MALDI-TOF MS) could also be employed given that they offer greater throughput in sample screening for viral pathogens. Molecular methods should be provided with an appropriate set of controls, including an internal amplification control, to confirm the validity of the results obtained.

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“…More recently, Dalton et al [121] developed a diagnostic real-time RT-PCR for the detection of RHDVb strains targeting a 91-bp amplicon within the VP60 protein that covers nucleotides (nts) 6190-6280. The RT-PCR is carried out as a duplex using the endogenous amplification control of the beta-actin gene from a commercial kit (EXOPOL S.L.).…”

Section: Rabbit Hemorrhagic Diseasementioning

confidence: 99%

“…Primers were designed to cover a 449-nt region from the 3′ region of the RHDV polymerase (3D) to the sg promoter region. Degenerate primers were designed to bind at positions 4837-4856 and 5266-5286 in the RHDV2 genome [121].…”

Section: Rabbit Hemorrhagic Diseasementioning

confidence: 99%

The Health and Future of the Six Hare Species in Europe: A Closer Look at the Iberian Hare

Duarte¹,

Carvalho²,

Santos³

et al. 2021

Lagomorpha Characteristics

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Although there are around 40 species of hares in the world divided into three different genera (Lepus, Caprolagus, and Pronolagus), only six species inhabit Europe, all belonging to genus Lepus. The conservation status of these six species was recently revised in the International Conservation Union (IUCN) Red List of Threatened Species. Lepus castroviejoi and L. corsicanus were attributed the status of "vulnerable". The other four species, L. europaeus, L. timidus, L. capensis, and L. granatensis, were considered of "least concern" although a declining trend was recognized for the last two species' wild populations. Here we review the major threats to the hare species in Europe, with emphasis on infectious diseases. Furthermore, we present the sanitary data regarding the Iberian hare populations from Portugal, which were severely affected by the emergence of a naturally occurring recombinant myxoma virus (MYXV), first reported in mid-2018. The recent detection in 2019 of a leporid herpesvirus (LeHV-5), which pathogenicity appears to be exacerbated in MYXV-infected hares, brings additional concerns to the health and conservation of the Iberian hare.

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Conventional and real time RT-PCR assays for the detection and differentiation of variant rabbit hemorrhagic disease virus (RHDVb) and its recombinants

Cited by 25 publications

References 27 publications

Rabbit Hemorrhagic Disease Virus Non-structural Protein 6 Induces Apoptosis in Rabbit Kidney Cells

Rabbit Hemorrhagic Disease Virus Non-structural Protein 6 Induces Apoptosis in Rabbit Kidney Cells

Molecular methods in detection and epidemiologic studies of rabbit and hare viruses: a review

The Health and Future of the Six Hare Species in Europe: A Closer Look at the Iberian Hare

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