BlueTYPE – A low density TaqMan-RT-qPCR array for the identification of all 24 classical Bluetongue virus serotypes

Ries, Christina; Beer, Martin

doi:10.1016/j.jviromet.2020.113881

Cited by 8 publications

(12 citation statements)

References 46 publications

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“…However, several outbreaks in regions located further north have been reported, mainly facilitated by the recent spread of insect vectors [ 3 , 4 , 5 , 6 ]. To date, 29 serotypes of BTV have been identified [ 7 ], with two more putative serotypes and several other variants being further described. At this time, the described serotypes of BTV are classified as typical, ranging from 1 to 24, recognized by cross-neutralization assays and confirmed by phylogenetic analysis [ 1 ], and atypical (25–27) [ 2 , 3 , 4 , 7 ], found exclusively in small ruminants, non-pathogenic and spread by direct contact transmission [ 8 , 9 , 10 , 11 ].…”

Section: Introductionmentioning

confidence: 99%

“…To date, 29 serotypes of BTV have been identified [ 7 ], with two more putative serotypes and several other variants being further described. At this time, the described serotypes of BTV are classified as typical, ranging from 1 to 24, recognized by cross-neutralization assays and confirmed by phylogenetic analysis [ 1 ], and atypical (25–27) [ 2 , 3 , 4 , 7 ], found exclusively in small ruminants, non-pathogenic and spread by direct contact transmission [ 8 , 9 , 10 , 11 ]. Additionally, serotypes BTV-28 and BTV-29, which are closely related to other typical BTV serotypes, have recently been described, and two others are being studied [ 4 , 12 , 13 ].…”

Section: Introductionmentioning

confidence: 99%

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Heterologous Combination of ChAdOx1 and MVA Vectors Expressing Protein NS1 as Vaccination Strategy to Induce Durable and Cross-Protective CD8+ T Cell Immunity to Bluetongue Virus

et al. 2020

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The sequence of non-structural protein NS1 of bluetongue virus (BTV), which contains immunodominant CD8+ T cell epitopes, is highly conserved among BTV serotypes, and has therefore become a major tool in the development of a universal BTV vaccine. In this work, we have engineered multiserotype BTV vaccine candidates based on recombinant chimpanzee adenovirus (ChAdOx1) and modified vaccinia virus Ankara (MVA) vectors expressing the NS1 protein of BTV-4 or its truncated form NS1-Nt. A single dose of ChAdOx1-NS1 or ChAdOx1-NS1-Nt induced a moderate CD8+ T cell response and protected IFNAR(-/-) mice against a lethal dose of BTV-4/MOR09, a reassortant strain between BTV-1 and BTV-4, although the animals showed low viremia after infection. Furthermore, IFNAR(-/-) mice immunized with a single dose of ChAdOx1-NS1 were protected after challenge with a lethal dose of BTV-8 in absence of viremia nor clinical signs. Additionally, the heterologous prime-boost ChAdOx1/MVA expressing NS1 or NS1-Nt elicited a robust NS1 specific CD8+ T cell response and protected the animals against BTV-4/MOR09 even 16 weeks after immunization, with undetectable levels of viremia at any time after challenge. Subsequently, the best immunization strategy based on ChAdOx1/MVA-NS1 was assayed in sheep. Non-immunized animals presented fever and viremia levels up to 104 PFU/mL after infection. In contrast, although viremia was detected in immunized sheep, the level of virus in blood was 100 times lower than in non-immunized animals in absence of clinical signs.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Heterologous Combination of ChAdOx1 and MVA Vectors Expressing Protein NS1 as Vaccination Strategy to Induce Durable and Cross-Protective CD8+ T Cell Immunity to Bluetongue Virus

et al. 2020

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“…The five BTV xMAP serotyping panels (A–E) represent the end result of an exhaustive optimization process which, we consider, will facilitate the implementation of the BTV xMAP assay in other laboratories. Recently, a low-density TaqMan-RT-qPCR array has been described for the classical BTV serotypes [ 43 ] and represents an approach to expedite BTV serotype determination. However, this assay utilizes numerous singleplex RT-qPCR assays, allowing for three samples to be fully serotyped on a 96-wellplate, which is less than what can be achieved using the BTV xMAP ( n = 12).…”

Section: Discussionmentioning

confidence: 99%

Simultaneous Detection of Bluetongue Virus Serotypes Using xMAP Technology

et al. 2020

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Bluetongue is an economically important disease of ruminants caused by the bluetongue virus (BTV). BTV is serologically diverse, which complicates vaccination strategies. Rapid identification of the causative BTV serotypes is critical, however, real-time PCR (RT-qPCR) can be costly and time consuming to perform when the circulating serotypes are unknown. The Luminex xMAP technology is a high-throughput platform that uses fluorescent beads to detect multiple targets simultaneously. We utilized existing BTV serotyping RT-qPCR assays for BTV-1 to BTV-24 and adapted them for use with the xMAP platform. The xMAP assay specifically detected all 24 BTV serotypes when testing reference strains. In all BTV-positive samples, the sensitivity of the BTV xMAP was 87.55% whereas the sensitivity of the serotype-specific RT-qPCR was 79.85%. The BTV xMAP assay allowed for the specific detection of BTV serotypes 1–24 at a lower cost than current RT-qPCR assays. Overall, the assay provides a useful novel diagnostic tool, particularly when analyzing large sample sets. The use of the BTV xMAP assay will allow for the rapid assessment of BTV epidemiology and may inform decision-making related to control and prevention measures.

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“…At all 5 blood sampling time points, EDTA blood was analyzed with the BTV-25 specific RT-qPCR (BTV-25-Mix13 assay). Furthermore, individual EDTA blood samples were tested in the BlueTYPE array as described previously [9].…”

Section: Rna Extraction and Rt-qpcrmentioning

confidence: 99%

“…Nevertheless, the OIE recommended the Pan-BTV-segment 10 RT-qPCR [7,8] in order to detect all BTV serotypes, including the known atypical BTVs. Recently, we established the Pan-BTV-Classic-S1-RT-qPCR assay, targeting BTV segment 1 for distinction between classical and atypical serotypes [9]. The discovery of TOV was followed by the description of BTV-26 in samples from symptomatic sheep in Kuwait [3].…”

Section: Introductionmentioning

confidence: 99%

Isolation and Cultivation of a New Isolate of BTV-25 and Presumptive Evidence for a Potential Persistent Infection in Healthy Goats

Ries

Domes²,

Janowetz³

et al. 2020

Viruses

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Recently, several so-called “atypical” Bluetongue virus (BTV) serotypes were discovered, including BTV-25 (Toggenburg virus), in Switzerland. Most “atypical” BTV were identified in small ruminants without clinical signs. In 2018, two goats from a holding in Germany tested positive for BTV-25 genome by RT-qPCR prior to export. After experimental inoculation of the two goats with the BTV-25 positive field blood samples for generation of reference materials, viremia could be observed in one animal. For the first time, the BTV-25-related virus was isolated in cell culture from EDTA-blood and the full genome of isolate “BTV-25-GER2018” could be generated. BTV-25-GER2018 was only incompletely neutralized by ELISA-positive sera. We could monitor the BTV-25 occurrence in the respective affected goat flock of approximately 120 goats over several years. EDTA blood samples were screened with RT-qPCR using a newly developed BTV-25 specific assay. For serological surveillance, serum samples were screened using a commercial cELISA. BTV-25-GER2018 was detected over 4.5 years in the goat flock with intermittent PCR-positivity in some animals, and with or without concomitantly detected antibodies since 2015. We could demonstrate the viral persistence of BTV-25-GER2018 in goats for up to 4.5 years, and the first BTV-25 isolate is now available for further characterization.

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BlueTYPE – A low density TaqMan-RT-qPCR array for the identification of all 24 classical Bluetongue virus serotypes

Cited by 8 publications

References 46 publications

Heterologous Combination of ChAdOx1 and MVA Vectors Expressing Protein NS1 as Vaccination Strategy to Induce Durable and Cross-Protective CD8+ T Cell Immunity to Bluetongue Virus

Heterologous Combination of ChAdOx1 and MVA Vectors Expressing Protein NS1 as Vaccination Strategy to Induce Durable and Cross-Protective CD8+ T Cell Immunity to Bluetongue Virus

Simultaneous Detection of Bluetongue Virus Serotypes Using xMAP Technology

Isolation and Cultivation of a New Isolate of BTV-25 and Presumptive Evidence for a Potential Persistent Infection in Healthy Goats

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