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

Abrantes, Joana; Lopes, Ana M.

doi:10.3390/microorganisms9050972

Cited by 11 publications

(6 citation statements)

References 169 publications

(375 reference statements)

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“…In short, a semi-purified preparation of each virus was then prepared from liver homogenates and inactivated using binary ethylenimine [ 22 ]. The antigen preparation, measured in hemagglutination (HA) units per mL [ 23 ], was diluted to the desired concentration in phosphate buffered saline and adjuvanted with an equal volume of Montanide ISA-201 (Seppic, Courbevoie, France). Rabbits were injected subcutaneously with either 1 mL of the G1.2-specific vaccine (representing a 100 HA dose), or 0.5 mL of the multivalent RHDV vaccine (consisting of a 32 HA dose of each genotype—GI.1a, GI.1c and GI.2).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Immunological Cross-Protection between Different Rabbit Hemorrhagic Disease Viruses—Implications for Rabbit Biocontrol and Vaccine Development

et al. 2022

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The use of rabbit hemorrhagic disease virus (RHDV) as a biocontrol agent to control feral rabbit populations in Australia, in combination with circulating endemic strains, provides a unique environment to observe the interactions between different lagoviruses competing for the same host. Following the arrival of RHDV2 (GI.2) in Australia, it became necessary to investigate the potential for immunological cross-protection between different variants, and the implications of this for biocontrol programs and vaccine development. Laboratory rabbits of various immune status—(1) rabbits with no detectable immunity against RHDV; (2) rabbits with experimentally acquired immunity after laboratory challenge; (3) rabbits immunised with a GI.2-specific or a multivalent RHDV inactivated virus prototype vaccine; or (4) rabbits with naturally acquired immunity—were challenged with one of three different RHDV variants (GI.1c, GI.1a or GI.2). The degree of cross-protection observed in immune rabbits was associated with the variant used for challenge, infectious dose of the virus and age, or time since acquisition of the immunity, at challenge. The immune status of feral rabbit populations should be determined prior to intentional RHDV release because of the high survival proportions in rabbits with pre-existing immunity. In addition, to protect domestic rabbits in Australia, a multivalent RHDV vaccine should be considered because of the limited cross-protection observed in rabbits given monovalent vaccines.

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Section: Methodsmentioning

confidence: 99%

“…To help clarify the immune status of individual rabbits, where cross-reactivity between the different antigens was suspected, samples were also tested in three separate hemagglutination inhibition (HI) tests against GI.1a, GI.1c and GI.2 antigens [ 23 ].…”

Section: Methodsmentioning

confidence: 99%

Immunological Cross-Protection between Different Rabbit Hemorrhagic Disease Viruses—Implications for Rabbit Biocontrol and Vaccine Development

et al. 2022

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“…Within one to three days after infection via oral or nasal route transmission in animals, rabbits manifest the first clinical signs, such as apathy, fever, and respiratory disorders [ 12 , 13 ]. The highest titer of L. europaeus /RHDV is found in the liver, spleen (reported in the presence of the chronic or sub-acute forms of the disease), lungs, kidneys, and bone marrow, which are the most frequently assessed organs in the diagnostic process [ 12 , 14 , 15 ]. L. europaeus /RHDV can also be detected in biological fluids such as serum, urine, and feces samples [ 15 ].…”

Section: Introductionmentioning

confidence: 99%

“…The highest titer of L. europaeus /RHDV is found in the liver, spleen (reported in the presence of the chronic or sub-acute forms of the disease), lungs, kidneys, and bone marrow, which are the most frequently assessed organs in the diagnostic process [ 12 , 14 , 15 ]. L. europaeus /RHDV can also be detected in biological fluids such as serum, urine, and feces samples [ 15 ]. During the development of RHD, many pathological changes occur in rabbit organs, especially in the liver (the site of viral replication), lungs, kidneys, spleen, and trachea [ 12 ].…”

Section: Introductionmentioning

confidence: 99%

Digital PCR (dPCR) Quantification of miR-155-5p as a Potential Candidate for a Tissue Biomarker of Inflammation in Rabbits Infected with Lagovirus europaeus/Rabbit Hemorrhagic Disease Virus (RHDV)

Hukowska-Szematowicz

Ostrycharz

Dudzińska

et al. 2023

Viruses

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MicroRNAs (miRNAs, miRs) are a group of small, 17–25 nucleotide, non-coding RNA sequences that, in their mature form, regulate gene expression at the post-transcriptional level. They participate in many physiological and pathological processes in both humans and animals. One such process is viral infection, in which miR-155 participates in innate and adaptive immune responses to a broad range of inflammatory mediators. Recently, the study of microRNA has become an interesting field of research as a potential candidate for biomarkers for various processes and disease. To use miRNAs as potential biomarkers of inflammation in viral diseases of animals and humans, it is necessary to improve their detection and quantification. In a previous study, using reverse transcription real-time quantitative PCR (RT-qPCR), we showed that the expression of ocu-miR-155-5p in liver tissue was significantly higher in rabbits infected with Lagovirus europaeus/Rabbit Hemorrhagic Disease Virus (RHDV) compared to healthy rabbits. The results indicated a role for ocu-miR-155-5p in Lagovirus europaeus/RHDV infection and reflected hepatitis and the impairment/dysfunction of this organ during RHD. MiR-155-5p was, therefore, hypothesized as a potential candidate for a tissue biomarker of inflammation and examined in tissues in Lagovirus europaeus/RHDV infection by dPCR. The objective of the study is the absolute quantification of ocu-miR-155-5p in four tissues (liver, lung, kidney, and spleen) of rabbits infected with Lagovirus europaeus/RHDV by digital PCR, a robust technique for the precise and direct quantification of small amounts of nucleic acids, including miRNAs, without standard curves and external references. The average copy number/µL (copies/µL) of ocu-miRNA-155-5p in rabbits infected with Lagovirus europaeus GI.1a/Rossi in the liver tissue was 12.26 ± 0.14, that in the lung tissue was 48.90 ± 9.23, that in the kidney tissue was 16.92 ± 2.89, and that in the spleen was 25.10 ± 0.90. In contrast, in the tissues of healthy control rabbits, the average number of copies/µL of ocu-miRNA-155-5p was 5.07 ± 1.10 for the liver, 23.52 ± 2.77 for lungs, 8.10 ± 0.86 for kidneys, and 42.12 ± 3.68 for the spleen. The increased expression of ocu-miRNA-155-5p in infected rabbits was demonstrated in the liver (a fold-change of 2.4, p-value = 0.0003), lung (a fold-change of 2.1, p-value = 0.03), and kidneys (a fold-change of 2.1, p-value = 0.01), with a decrease in the spleen (a fold-change of 0.6, p-value = 0.002). In the study of Lagovirus europaeus/RHDV infection and in the context of viral infections, this is the first report that shows the potential use of dPCR for the sensitive and absolute quantification of microRNA-155-5p in tissues during viral infection. We think miR-155-5p may be a potential candidate for a tissue biomarker of inflammation with Lagovirus europaeus/RHDV infection. Our report presents a new path in discovering potential candidates for the tissue biomarkers of inflammation.

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“…Regarding GI.1 or GI.2 detection, several techniques of different complexity can be employed, such as immunohistochemistry, next-generation sequencing or RT-PCR (reviewed in [32]). Moreover, some commercial and non-commercial ELISA tests for which reagents are sold have been described for antigen detection [2,33,34].…”

Section: Introductionmentioning

confidence: 99%

Development and Evaluation of a Duplex Lateral Flow Assay for the Detection and Differentiation between Rabbit Haemorrhagic Disease Virus Lagovirus europaeus/GI.1 and /GI.2

Fresco-Taboada

Montón

Tapia

et al. 2022

Biology

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Rabbit Haemorrhagic Disease Virus 2 (RHDV2, recently named Lagovirus europaeus/GI.2) was first reported in France in 2010 and has spread globally since then, replacing most of the circulating former RHDV (genotype GI.1) in many countries. The detection and differentiation of both genotypes is of crucial importance for the surveillance of the disease. In this article, a duplex lateral flow assay (LFA) for antigen detection is described and evaluated, providing the first description of a quick and easy-to-use test that allows for the simultaneous detection and differentiation of RHDV genotypes GI.1 and GI.2. A panel of GI.1- or GI.2-infected and non-infected rabbit liver samples and liver exudates (136 samples) was analysed, obtaining a total sensitivity of 94.4% and specificity of 100%. These data confirm that the developed duplex LFA can be used as a reliable diagnostic test for RHD surveillance, especially in farms and the field.

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

Cited by 11 publications

References 169 publications

Immunological Cross-Protection between Different Rabbit Hemorrhagic Disease Viruses—Implications for Rabbit Biocontrol and Vaccine Development

Immunological Cross-Protection between Different Rabbit Hemorrhagic Disease Viruses—Implications for Rabbit Biocontrol and Vaccine Development

Digital PCR (dPCR) Quantification of miR-155-5p as a Potential Candidate for a Tissue Biomarker of Inflammation in Rabbits Infected with Lagovirus europaeus/Rabbit Hemorrhagic Disease Virus (RHDV)

Development and Evaluation of a Duplex Lateral Flow Assay for the Detection and Differentiation between Rabbit Haemorrhagic Disease Virus Lagovirus europaeus/GI.1 and /GI.2

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