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

Wilson, William C.; Hindson, Benjamin J.; O'Hearn, Emily S.; Hall, Steven G.; Tellgren-Roth, Christian; Torres, Clinton; Naraghi-Arani, Pejman; Mecham, James O.; Lenhoff, Raymond J.

doi:10.1177/104063870902100602

Cited by 34 publications

(24 citation statements)

References 41 publications

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“…There are fewer real-time PCR assays available for EHDV (Wilson et al 2009, Clavijo et al 2010, Eschbaumer et al 2012). These PCR-based diagnostic procedures are exquisitely analytically sensitive and theoretically can be configured to any desired specificity if the sequence of the target viral gene of interest is known.…”

Section: Nucleic Acid Detectionmentioning

confidence: 98%

“…Detection of viral RNA by conventional or real-time RT-PCR (real-time PCR) is useful for screening samples prior to virus isolation, with the latter being the preferred method. There are published and/or commercial real-time PCR tests for both of these viruses ( Jimenez-Clavero et al 2006, Shaw et al 2007, Toussaint et al 2007, Wilson et al 2009, Hoffmann et al 2009, Clavijo et al 2010, Schroeder et al 2013). There is a conventional single-tube multiplex RT-PCR assay for determining the serotype of the primary US BTV strains ( Johnson et al 2000), as well as an array of conventional RT-PCR assays able to identify and differentiate between the 26 known BTV serotypes (Maan et al 2012).…”

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confidence: 95%

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Diagnostic Tools for Bluetongue and Epizootic Hemorrhagic Disease Viruses Applicable to North American Veterinary Diagnosticians

Wilson

Daniels

Ostlund

et al. 2015

Vector-Borne and Zoonotic Diseases

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This review provides an overview of current and potential new diagnostic tests for bluetongue (BT) and epizootic hemorrhagic disease (EHD) viruses compiled from international participants of the Orbivirus Gap Analysis Workshop, Diagnostic Group. The emphasis of this review is on diagnostic tools available to North American veterinary diagnosticians. Standard diagnostic tests are readily available for BT/EHD viruses, and there are described tests that are published in the World Organization for Animal Health (OIE) Terrestrial Manual. There is however considerable variation in the diagnostic approach to these viruses. Serological assays are well established, and many laboratories are experienced in running these assays. Numerous nucleic acid amplification assays are also available for BT virus (BTV) and EHD virus (EHDV). Although there is considerable experience with BTV reverse-transcriptase PCR (RT-PCR), there are no standards or comparisons of the protocols used by various state and federal veterinary diagnostic laboratories. Methods for genotyping BTV and EHDV isolates are available and are valuable tools for monitoring and analyzing circulating viruses. These methods include RT-PCR panels or arrays, RT-PCR and sequencing of specific genome segments, or the use of next-generation sequencing. In addition to enabling virus characterization, use of advanced molecular detection methods, including DNA microarrays and next-generation sequencing, significantly enhance the ability to detect unique virus strains that may arise through genetic drift, recombination, or viral genome segment reassortment, as well as incursions of new virus strains from other geographical areas.

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Section: Nucleic Acid Detectionmentioning

confidence: 98%

mentioning

confidence: 95%

Diagnostic Tools for Bluetongue and Epizootic Hemorrhagic Disease Viruses Applicable to North American Veterinary Diagnosticians

Wilson

Daniels

Ostlund

et al. 2015

Vector-Borne and Zoonotic Diseases

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“…Notably, a leucine residue is found at position 24 of the majority of BTV NS3 sequences available in the database, but among those belonging to the Western group 1 topotype, P24 is not unique to BTV-2RSA. Indeed, P24 also is present in the NS3 protein sequences of at least three BTV-2 (isolated from India or Italy) and two viruses of other serotypes (BTV-11 from the United States and BTV-12 from Jamaica) (51,(80)(81)(82). Interestingly, NS3/NS3a bearing P24 instead of L24 appears very unstable in ovine but not in Culicoides cells, most likely due to a more efficient and host-specific protein degradation.…”

Section: Discussionmentioning

confidence: 99%

Turnover Rate of NS3 Proteins Modulates Bluetongue Virus Replication Kinetics in a Host-Specific Manner

Ftaich

Ciancia

Viarouge

et al. 2015

J Virol

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Bluetongue virus (BTV) is an arbovirus transmitted to livestock by midges of the Culicoides family and is the etiological agent of a hemorrhagic disease in sheep and other ruminants. In mammalian cells, BTV particles are released primarily by virus-induced cell lysis, while in insect cells they bud from the plasma membrane and establish a persistent infection. BTV possesses a ten-segmented double-stranded RNA genome, and NS3 proteins are encoded by segment 10 (Seg-10). The viral nonstructural protein 3 (NS3) plays a key role in mediating BTV egress as well as in impeding the in vitro synthesis of type I interferon in mammalian cells. In this study, we asked whether genetically distant NS3 proteins can alter BTV-host interactions. Using a reverse genetics approach, we showed that, depending on the NS3 considered, BTV replication kinetics varied in mammals but not in insects. In particular, one of the NS3 proteins analyzed harbored a proline at position 24 that leads to its rapid intracellular decay in ovine but not in Culicoides cells and to the attenuation of BTV virulence in a mouse model of disease. Overall, our data reveal that the genetic variability of Seg-10/NS3 differentially modulates BTV replication kinetics in a host-specific manner and highlight the role of the host-specific variation in NS3 protein turnover rate. IMPORTANCEBTV is the causative agent of a severe disease transmitted between ruminants by biting midges of Culicoides species. NS3, encoded by Seg-10 of the BTV genome, fulfills key roles in BTV infection. As Seg-10 sequences from various BTV strains display genetic variability, we assessed the impact of different Seg-10 and NS3 proteins on BTV infection and host interactions. In this study, we revealed that various Seg-10/NS3 proteins alter BTV replication kinetics in mammals but not in insects. Notably, we found that NS3 protein turnover may vary in ovine but not in Culicoides cells due to a single amino acid residue that, most likely, leads to rapid and host-dependent protein degradation. Overall, this study highlights that genetically distant BTV Seg-10/NS3 influence BTV biological properties in a host-specific manner and increases our understanding of how NS3 proteins contribute to the outcome of BTV infection. Bluetongue virus (BTV) is an arthropod-borne virus responsible for a hemorrhagic disease of domestic and wild ruminants (1, 2) and is transmitted between mammalian hosts by biting midges of the Culicoides family (3, 4). C. imicola represents the major vector species in Africa (3) and in southern Europe (5, 6), whereas C. obsoletus and C. pulicaris species have been involved in the transmission of BTV in central and northern Europe (7-9). The impact of bluetongue disease in Europe was relatively low before the 1990s (6, 10, 11). However, since 1998, several seasonal incursions of distinct BTV serotypes/strains have occurred virtually every year, causing massive economic losses to animal husbandry (7,(12)(13)(14)(15)(16). BTV is a nonenveloped virus that belongs to the Orbi...

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“…However, Seg-5/NS1 of IND2003/08 showed up to 99% identity with western topotype viruses (prototype 600565 strain), providing further evidence for the introduction of western BTV strains and reassortment between eastern and western field strains in India (6,9,23). Western topotype or reassortant BTV strains may be at least partly responsible for the increased virulence of bluetongue outbreaks seen in India, in indigenous sheep breeds.…”

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confidence: 85%

The Genome Sequence of a Reassortant Bluetongue Virus Serotype 3 from India

Maan

Guimera

et al. 2012

J Virol

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All 10 genome segments (Seg-1 to 10 -a total of 19,188 bp) were sequenced from a strain of bluetongue virus serotype 3 (BTV-3) from India (strain IND2003/08). Sequence comparisons showed that nine of the genome segments from this virus group with other eastern topotype strains. Genome Seg-2 and Seg-6 group with eastern BTV-3 strains from Japan. However, Seg-5 (the NS1 gene) from IND2003/08 belongs to a western lineage, demonstrating that IND2003/08 is a reassortant between eastern and western topotype bluetongue viruses. This confirms that western BTV strains have been imported and are circulating within the subcontinent.

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

Cited by 34 publications

References 41 publications

Diagnostic Tools for Bluetongue and Epizootic Hemorrhagic Disease Viruses Applicable to North American Veterinary Diagnosticians

Diagnostic Tools for Bluetongue and Epizootic Hemorrhagic Disease Viruses Applicable to North American Veterinary Diagnosticians

Turnover Rate of NS3 Proteins Modulates Bluetongue Virus Replication Kinetics in a Host-Specific Manner

The Genome Sequence of a Reassortant Bluetongue Virus Serotype 3 from India

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