An improved Real-Time Polymerase Chain Reaction for the Simultaneous Detection of All Serotypes of <i>Epizootic Hemorrhagic Disease Virus</i>

Clavijo, Alfonso; Sun, Feng; Lester, Thomas; Jasperson, Dane C.; Wilson, William C.

doi:10.1177/104063871002200414

Cited by 32 publications

(24 citation statements)

References 21 publications

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“…A previously described rRT-PCR method was used to detect EHDV (Clavijo et al, 2010). Using the sequence for VP7, a 59-nuclease probe was designed to detect the mobuck virus.…”

Section: E Cooper and Othersmentioning

confidence: 99%

Mobuck virus genome sequence and phylogenetic analysis: identification of a novel Orbivirus isolated from a white-tailed deer in Missouri, USA

Cooper

Anbalagan

Klumper

et al. 2014

Journal of General Virology

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The genus Orbivirus includes a diverse group of segmented dsRNA viruses that are transmitted via arthropods, have a global distribution and affect a wide range of hosts. A novel orbivirus was co-isolated with epizootic haemorrhagic disease virus (EHDV) from a white-tailed deer (Odocoileus virginianus) exhibiting clinical signs characteristic of EHDV. Using antiserum generated against EHDV, a pure isolate of the novel non-cytopathic orbivirus was obtained in Aedes albopictus cell culture. Genomic sequencing and phylogenetic analysis of predicted ORFs showed that eight of the ten ORFs were most homologous to Peruvian horse sickness virus (PHSV), with amino acid identities of 44.3-73.7 %. The remaining two ORFs, VP3 and VP5, were most similar to Middle Point orbivirus (35.9 %) and Yunnan orbivirus (59.8 %), respectively. Taxonomic classification of orbiviruses is largely based on homology of the major subcore structural protein VP2(T2), encoded by segment 2 for mobuck virus. With only 69.1 % amino acid identity to PHSV, we propose mobuck virus as the prototype of a new species of Orbivirus.

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“…A previously described rRT-PCR method was used to detect EHDV (Clavijo et al, 2010). Using the sequence for VP7, a 59-nuclease probe was designed to detect the mobuck virus.…”

Section: E Cooper and Othersmentioning

confidence: 99%

Mobuck virus genome sequence and phylogenetic analysis: identification of a novel Orbivirus isolated from a white-tailed deer in Missouri, USA

Cooper

Anbalagan

Klumper

et al. 2014

Journal of General Virology

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“…Quantitative PCR assays are the preferred strategy, as they are for BTV (93), since the amount of virus in an individual sample can be estimated, and the risks of laboratory contamination are reduced. Both serogroup-specific and serotype-specific real-time PCR assays are now available to quickly identify and serotype any EHDV contained within a biological sample (4,69,74,94).…”

Section: Diagnosismentioning

confidence: 99%

Epizootic haemorrhagic disease

Maclachlan¹,

Zientara²,

Savini³

et al. 2015

Rev. Sci. Tech. OIE

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SummaryEpizootic haemorrhagic disease (EHD) is an arthropod-transmitted viral disease of certain wild ungulates, notably North American white-tailed deer and, more rarely, cattle. The disease in white-tailed deer results from vascular injury analogous to that caused by bluetongue virus (BTV), to which EHD virus (EHDV) is closely related. There are seven serotypes of EHDV recognised, and Ibaraki virus, which is the cause of sporadic disease outbreaks in cattle in Asia, is included in EHDV serotype 2. The global distribution and epidemiology of BTV and EHDV infections are also similar, as both viruses occur throughout temperate and tropical regions of the world where they are transmitted by biting Culicoides midges and infect a wide variety of domestic and wild ungulates. However, the global distribution and epidemiology of EHDV infection are less well characterised than they are for BTV. Whereas most natural and experimental EHDV infections (other than Ibaraki virus infection) of livestock are subclinical or asymptomatic, outbreaks of EHD have recently been reported among cattle in the Mediterranean Basin, Reunion Island, South Africa, and the United States. Accurate and convenient laboratory tests are increasingly available for the sensitive and specific serological and virological diagnosis of EHDV infection and confirmation of EHD in animals, but commercial vaccines are available only for prevention of Ibaraki disease and not for protection against other strains and serotypes of EHDV.

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“…The translated product of the segment 2 gene (VP2) is considered as the major serotype-specific antigen (Anthony et al 2009). A TaqMan real-time reverse transcriptase (RT)-PCR capable of detecting all eight serotypes of EHDV has been described (Clavijo et al 2010); however, the assay was not designed to differentiate the most common North American serotypes of EHDV. Identification of the specific serotypes of EHDV in clinical samples is important to better understand the epidemiology of the disease and to facilitate the development and use of vaccines.…”

mentioning

confidence: 99%

“…We collected tissue (spleen, lung, or lymphatic tissue) or ethylenediaminetetraacetic acid (EDTA) blood samples from white-tailed deer, submitted to the Texas A&M Veterinary Medical Diagnostic Laboratory (TVMDL, College Station, Texas, USA), and found EHDV positive by TaqMan real-time RT-PCR (Clavijo et al 2010). Viral RNA was extracted by a highthroughput Kingfisher 96 magnetic particle processor (Thermo Scientific, Waltham, Massachusetts, USA) using the Ambion MagMax RNA viral isolation kit (AM1836; Life Technologies, Carlsbad, California, USA).…”

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confidence: 99%

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Molecular Typing of Epizootic Hemorrhagic Disease Virus Serotypes by One-Step Multiplex RT-PCR

Sun

Cochran

Beckham

et al. 2014

Journal of Wildlife Diseases

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ABSTRACT:Epizootic hemorrhagic disease virus (EHDV) causes a highly infectious noncontagious hemorrhagic disease in wild and captive deer (Cervidae) populations in the US. Although rapid and accurate identification of the disease is important, identification of the serotype is equally important for understanding the epidemiology of the disease in white-tailed deer (Odocoileus virginianus) populations. We developed a one-step multiplex reverse transcriptase PCR assay for rapid differentiation and identification of EHDV serotypes 1, 2, and 6 in cell culture and clinical samples by targeting the viral gene segment 2 (L2) that encodes for the structural protein VP2. From 2009 to 2012, 427 clinical samples including tissue and blood (in ethylenediaminetetraacetic acid) from white-tailed deer, found EHDV positive by real-time PCR, were used to evaluate this subtyping assay. Eighteen percent of the positive samples tested were EHDV-1, 59% were EHDV-2, and 21% were EHDV-6; 2% of the samples were positive for more than one subtype, indicating mixed infection. This assay provides a rapid, sensitive, specific diagnostic tool for differentiation and identification of EHDV serotypes in field samples and virus isolates.

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An improved Real-Time Polymerase Chain Reaction for the Simultaneous Detection of All Serotypes of Epizootic Hemorrhagic Disease Virus

Cited by 32 publications

References 21 publications

Mobuck virus genome sequence and phylogenetic analysis: identification of a novel Orbivirus isolated from a white-tailed deer in Missouri, USA

Mobuck virus genome sequence and phylogenetic analysis: identification of a novel Orbivirus isolated from a white-tailed deer in Missouri, USA

Epizootic haemorrhagic disease

Molecular Typing of Epizootic Hemorrhagic Disease Virus Serotypes by One-Step Multiplex RT-PCR

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