Development and initial evaluation of a real-time RT-PCR assay to detect bluetongue virus genome segment 1

Shaw, Andrew; Monaghan, Paul; Alpar, H. Oya; Anthony, Simon J.; Darpel, Karin E.; Batten, Carrie; Guercio, Annalisa; Vitale, María; Bankowska, K.; Carpenter, Simon; Jones, Helen G.R.; Oura, Christopher; King, Donald P.; Elliott, Heather G.; Mellor, Philip A.; Mertens, Peter P. C.

doi:10.1016/j.jviromet.2007.05.014

Cited by 139 publications

(114 citation statements)

References 34 publications

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“…12 A number of real-time RT-PCR assays were recently developed for BTV. 7,14,29,32,36 The genome segments are commonly denoted as L1-3, M4-6, and S7-10, based on sequence size, similar to the Reoviridae nomenclature. The coding assignments are based on sequence size and protein produced, with structural proteins denoted as VP1-7 and nonstructural protein denoted as NS1-3.…”

Section: Introductionmentioning

confidence: 99%

A Multiplex Real-Time Reverse Transcription Polymerase Chain Reaction Assay for Detection and Differentiation of Bluetongue Virus and Epizootic Hemorrhagic Disease Virus Serogroups

et al. 2009

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Abstract. Bluetongue virus (BTV) causes disease in domestic and wild ruminants and results in significant economic loss. The closely related Epizootic hemorrhagic disease virus (EHDV) has been associated with bluetongue-like disease in cattle. Although U.S. EHDV strains have not been experimentally proven to cause disease in cattle, there is serologic evidence of infection in cattle. Therefore, rapid diagnosis and differentiation of BTV and EHDV is required. The genetic sequence information and bioinformatic analysis necessary to design a real-time reverse transcription polymerase chain reaction (RT-PCR) assay for the early detection of indigenous and exotic BTV and EHDV is described. This sequence data foundation focused on 2 conserved target genes: one that is highly expressed in infected mammalian cells, and the other is highly expressed in infected insect cells. The analysis of all BTV and EHDV prototype strains indicated that a complex primer design was necessary for both a virus group-comprehensive and virus group-specific gene amplification diagnostic test. This information has been used as the basis for the development of a rapid multiplex BTV-EHDV real-time RT-PCR that detects all known serotypes of both viruses and distinguishes between BTV and EHDV serogroups. The sensitivity of this rapid, single-tube, real-time RT-PCR assay is sufficient for diagnostic application, without the contamination problems associated with standard gel-based RT-PCR, especially nested RT-PCR tests.

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Section: Introductionmentioning

confidence: 99%

A Multiplex Real-Time Reverse Transcription Polymerase Chain Reaction Assay for Detection and Differentiation of Bluetongue Virus and Epizootic Hemorrhagic Disease Virus Serogroups

et al. 2009

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“…However, levels of antibody and viral RNA were unexpectedly low. Furthermore, presence of BTV RNA could not be confi rmed by use of other BTV-specifi c rRT-PCR protocols (13,14), and amplifi cation curves obtained in the screening rRT-PCR suggested that the target sequence on RNA segment 10 of this virus might be different from all known BTV strains (data not shown). For this reason, we determined the nucleotide sequence of the rRT-PCR amplifi cation product.…”

mentioning

confidence: 97%

Genetic Characterization of Toggenburg Orbivirus, a New Bluetongue Virus, from Goats, Switzerland

Hofmann¹,

Renzullo²,

Mader³

et al. 2008

Emerg. Infect. Dis.

274

221

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A novel bluetongue virus (BTV) termed Toggenburg orbivirus (TOV) was detected in goats from Switzerland by using real-time reverse transcription-PCR. cDNA corresponding to the complete sequence of 7 of 10 double-stranded RNA segments of the viral genome was amplifi ed by PCR and cloned into a plasmid vector. Five clones for each genome segment were sequenced to determine a consensus sequence. BLAST analysis and dendrogram construction showed that TOV is closely related to BTV, although some genome segments are distinct from the 24 known BTV serotypes. Maximal sequence identity to any BTV ranged from 63% (segment 2) to 79% (segments 7 and 10). Because the gene encoding outer capsid protein 2 (VP2), which determines the serotype of BTV, is placed within the BTV serogroup, we propose that TOV represents an unknown 25th serotype of BTV.

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“…rRT-PCR assays have also been reported for the detection of other viruses which cause vesicular disease of livestock including swine vesicular disease (SVD) (29), vesicular stomatitis (VS) (11,28) and vesicular exanthema of swine (VES) (31) or symptomatic look-alike diseases including bluetongue (13,27,33), bovine viral diarrhea…”

Section: Introductionmentioning

confidence: 99%

Diagnostic evaluation of a multiplexed RT-PCR microsphere array assay for the detection of foot-and-mouth and look-alike disease viruses

Hindson¹,

Baker²,

Tammero³

et al. 2007

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A high-throughput multiplexed assay (Multiplex Version 1.0) was developed for the differential laboratory diagnosis of foot-and-mouth disease virus (FMDV) from viruses which cause clinically similar diseases of livestock. This assay simultaneously screens for five RNA and two DNA viruses using multiplexed reverse transcription PCR (mRT-PCR) amplification coupled with a microsphere hybridization array and flow-cytometric detection. Two of the seventeen primer-probe sets included in this multiplex assay were adopted from previously characterized

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Development and initial evaluation of a real-time RT-PCR assay to detect bluetongue virus genome segment 1

Cited by 139 publications

References 34 publications

A Multiplex Real-Time Reverse Transcription Polymerase Chain Reaction Assay for Detection and Differentiation of Bluetongue Virus and Epizootic Hemorrhagic Disease Virus Serogroups

A Multiplex Real-Time Reverse Transcription Polymerase Chain Reaction Assay for Detection and Differentiation of Bluetongue Virus and Epizootic Hemorrhagic Disease Virus Serogroups

Genetic Characterization of Toggenburg Orbivirus, a New Bluetongue Virus, from Goats, Switzerland

Diagnostic evaluation of a multiplexed RT-PCR microsphere array assay for the detection of foot-and-mouth and look-alike disease viruses

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