Development of a real time polymerase chain reaction assay for equine encephalosis virus

Rathogwa, N.M.; Quan, Melvyn; Smit, J.Q.; Lourens, Carina W.; Guthrie, Alan John; Vuuren, Moritz Van

doi:10.1016/j.jviromet.2013.10.018

Cited by 7 publications

(7 citation statements)

References 23 publications

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“…This test provides the means for a differential diagnosis between EEV and AHSV. A real-time PCR assay targeting genome segment 7 of EEV was also recently developed (15). The assay uses group-specific EEV primers (EEVVP7_F0028_ 0048: 5′-GATAGCGGCTAGAGCCCTTTC-3′ and EEVVP7_ R0085_0106: 5′-AACTTGAGGAGCCATRGTAGCT-3′) and a TaqMan ® MGB ™ hydrolysis probe (EEVVP7_P0054_ 0072_MGBTM probe: 5′-TAAGAGCATGTGTTACTGC-3′) to allow the detection of a median tissue culture infective dose (TCID 50 ) as small as 10 2.9 TCID 50 /ml of blood (with 95% confidence).…”

Section: Diagnostic Assays For Equine Encephalosis Virusmentioning

confidence: 99%

Zoonotic and emerging orbivirus infections

Attoui¹,

Jaafar²

2015

Rev. Sci. Tech. OIE

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Many novel emerging orbiviruses have been isolated in the past 15 years. Important viruses include Peruvian horse sickness virus (PHSV) and Yunnan orbivirus (YUOV), pathogens of equids which were originally isolated almost simultaneously from 1997 to 1999 in the People's Republic of China, Australia and Peru. YUOV has also been isolated from cattle, sheep and a dog. The isolation of YUOV from a dog is not the first case of an orbivirus being isolated from a carnivore. Bluetongue virus and African horse sickness virus were earlier detected in carnivores which fed on contaminated meat. PHSV and YUOV both offer an opportunity to study the emergence of a single pathogen in geographically distant locations, although the original point of emergence is still unidentified. PHSV has been isolated from horses with neurological disease both in Australia and in Peru (where it is now endemic). Serological and molecular diagnostic assays have been developed for these viruses to assist in their identification and diagnosis. Other orbiviruses, such as Palyam virus and Equine encephalosis virus, have more recently been identified outside their geographical boundaries and may represent a threat to domesticated livestock and horses, respectively. The article also reviews four zoonotic orbivirus species (Corriparta virus, Changuinola virus, Kemerovo virus and Orungo virus) which have been identified in livestock and/or wildlife.

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Section: Diagnostic Assays For Equine Encephalosis Virusmentioning

confidence: 99%

Zoonotic and emerging orbivirus infections

Attoui¹,

Jaafar²

2015

Rev. Sci. Tech. OIE

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“…For well-characterised orbiviruses such as BTV, EHDV and AHSV, serotype-specific qRT-PCR diagnostic tests have been successfully developed based on a highly variable genome Seg-2 (encoding the serotype-specific VP2 protein), and in many reference laboratories, these assays have become the first choice for diagnostic testing of 'suspect' clinical samples [25,26,42]. However, a virus species-specific qRT-PCR assay that was recently developed for EEV targeting, the conserved sequences of Seg-7 has comparatively low efficiency (81%) [35].…”

Section: Discussionmentioning

confidence: 99%

“…These methods, which can detect and identify the dsRNA viral genome by conventional gel-based reverse transcription polymerase chain reactions (RT-PCR) [27]; real-time RT-PCR (qRT-PCR) assays [29,31]; reverse transcription loop-mediated isothermal amplification (RT-LAMP) [32]; sequence analyses and phylogenetic comparisons [33,34] are more sensitive and more specific than serological diagnostic techniques and can provide an accurate quantitative assessment of the relatedness of different viruses. Although a qRT-PCR has recently been developed for detection of EEV genome Seg-7 [35], no qRT-PCR assays are as yet available for the identification of EEV serotype.…”

Section: Introductionmentioning

confidence: 99%

Quantitative RT-PCR assays for identification and typing of the Equine encephalosis virus

et al. 2018

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Equine encephalosis (EE) is an acute, arthropod-borne, noncontagious, febrile disease of equids. The clinical signs of EE are similar to milder forms of African horse sickness (AHS) and the two diseases can be easily confused. The Equine encephalosis virus (EEV) is a distinct virus species within the genus Orbivirus, family Reoviridae, with ten linear segments of dsRNA genome. Seven distinct serotypes of EEV have been recognised on the basis of sequence analyses of Seg-2. The need for differential diagnosis of similar forms of EE and AHS warranted the development of molecular diagnostic methods for specific detection and identification of EEV. We report the development of quantitative real-time RT-PCR assay for detection of any member of the EEV species targeting the highly conserved EEV Seg-9. Similar serotype-specific qRT-PCR assays were designed for each of the seven EEV serotypes targeting genome Seg-2, encoding the serotype determining VP2 protein. These assays were evaluated using different EEV serotypes and other closely related orbiviruses. They were shown to be EEV virus species-specific, or EEV type-specific capable of detecting 1 to 13 copies of viral RNA in clinical samples. The assays failed to detect RNA from closely related orbiviruses, including AHSV and Peruvian horse sickness virus (PHSV) isolates.

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“…The differences in prevalence and rate of exposure to individual serotypes within and between regions in South Africa may be attributed to the differences in their spatial and temporal distribution of certain Culicoides species, in combination with the differences in the competence of these vectors to specific serotypes. C. imicola is the main species in the northern regions of South Africa, which correspond with NS3 gene cluster B, while C. bolitinos is more abundant in the southern districts, corresponding with NS3 gene cluster A [5,20,38]. In addition, the low EEV seroprevalence in the Western Cape Province in South Africa could be attributed to the lower abundance of C. imicola in this region [5].…”

Section: Culicoides Sppmentioning

confidence: 98%

Equine Encephalosis Virus

Tirosh-Levy

Steinman

2022

Animals

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Equine encephalosis (EE) is an arthropod-borne, noncontagious, febrile disease of horses. It is caused by EE virus (EEV), an Orbivirus of the Reoviridae family transmitted by Culicoides. Within the EEV serogroup, seven serotypes (EEV-1–7) have been identified to date. This virus was first isolated from a horse in South Africa in 1967 and until 2008 was believed to be restricted to southern Africa. In 2008–2009, isolation of EEV in an outbreak reported from Israel demonstrated the emergence of this pathogen into new niches. Indeed, testing in retrospect sera samples revealed that EEV had already been circulating outside of South Africa since 2001. Although EEV normally does not cause severe clinical disease, it should be considered important since it may indicate the possible spread of other related, much more pathogenic viruses, such as African horse sickness virus (AHSV). The spread of EEV from South Africa to central Africa, the Middle East and India is an example of the possible emergence of new pathogens in new niches, as was seen in the case of West Nile virus, and should be a reminder not to limit the differential list when facing a possible outbreak or a cluster of clinical cases. This review summarizes current knowledge regarding EEV structure, pathogenesis, clinical significance, and epidemiology.

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Development of a real time polymerase chain reaction assay for equine encephalosis virus

Cited by 7 publications

References 23 publications

Zoonotic and emerging orbivirus infections

Zoonotic and emerging orbivirus infections

Quantitative RT-PCR assays for identification and typing of the Equine encephalosis virus

Equine Encephalosis Virus

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