A duplex real-time RT-PCR for the detection of bluetongue virus in bovine semen

Vanbinst, T.; Vandenbussche, Frank; Dernelle, Eric; Clercq, Kris De

doi:10.1016/j.jviromet.2010.07.019

Cited by 33 publications

(46 citation statements)

References 26 publications

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“…The results for the S5 assay, based on the same primers and probe used in the assay optimized for the screening of semen from bulls infected with BTV-8, 18,19 provides a good example of this variability. Even though it was the second best assay and detected many isolates with similar sensitivity to the S10 assay, there were instances where differences up to 6.6 Ct (almost 2 log 10 ) were observed.…”

Section: 10mentioning

confidence: 99%

Longitudinal study of the detection of Bluetongue virus in bull semen and comparison of real-time polymerase chain reaction assays

Davis

Walsh

et al. 2014

J VET Diagn Invest

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Abstract. Infection with Bluetongue virus (BTV) is a significant impediment to the global movement of bovine semen. Repeat testing of blood from donor animals is specified in the World Organization for Animal Health (OIE) Manual for the export of semen from regions where BTV may be present. Screening of blood or semen samples has usually been carried out by virus isolation (VI) either by inoculation of chicken embryos followed by passage onto insect and mammalian cell cultures or in vivo inoculation of sheep followed by serology to detect seroconversion. Direct testing of semen for BTV would enable earlier release of semen samples and avoid repeat testing of the donor, as well as provide an option for releasing batches of semen that were collected without certification of the donor. Quantitative (real-time) reverse transcription polymerase chain reaction (qRT-PCR) assays overcome most of the limitations of other methods and have the potential to provide higher sensitivity. The present study compared 5 qRT-PCR assays, including 2 commercially available kits, for the detection of BTV in semen serially collected from 8 bulls over a period of 90 days after experimental infection. The results of the study show that at least one of the qRT-PCR assays is extremely reproducible and has both very high sensitivity and specificity to reliably detect all available serotypes. The preferred qRT-PCR gave consistently superior results to VI, sheep inoculation, and conventional RT-PCR. Therefore, the assay can be recommended for the screening of bovine semen for freedom from BTV.

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Section: 10mentioning

confidence: 99%

Longitudinal study of the detection of Bluetongue virus in bull semen and comparison of real-time polymerase chain reaction assays

Davis

Walsh

et al. 2014

J VET Diagn Invest

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“…A direct identification of BTV in blood or tissue samples is possible with use of the reverse transcription-polymerase chain reaction (RT-PCR) method that allows for serotyping and can detect BTV RNA in samples as late as 6 months after infection (Sperlova and Zendulkova, 2011). A quantitative assessment of RNA in a sample is possible by real time-RT-PCR Toussaint et al, 2007;Vanbinst et al, 2010;De Leeuw et al, 2013). RNA polyacrylamide gel electrophoresis (PAGE) has been used as a diagnostic tool for the identification BTV 10 segments.…”

Section: Antigen Identificationmentioning

confidence: 99%

“…So far the real-time RT-PCR assays have not been validated to the level required by the OIE, although ring trials have been conducted . Vanbinst et al (2010) …”

Section: Real Time Rt-pcr (Qpcr)mentioning

confidence: 99%

Bluetongue: Virus proteins and recent diagnostic approaches

Bitew¹,

Sukdeb²,

Ravishankar³

2013

Afr. J. Microbiol. Res.

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“…The BTV-8 strain also appears to have had an increased tendency to be secreted in the semen of infected rams and bulls. Two studies have demonstrated the presence of the BTV-8 RNA and/or infectious virus in the semen of naturally infected bulls and rams by using PCR and/or virus isolation (Leemans et al, 2011;Vanbinst et al, 2010). In one of the studies 75-100% of semen samples collected from rams between 25-116 days post observation of clinical signs still tested positive for the virus by PCR (Leemans et al, 2011).…”

Section: Why Is It Important To Identify the Molecular Determinants Tmentioning

confidence: 99%

Bluetongue virus genetic and phenotypic diversity: Towards identifying the molecular determinants that influence virulence and transmission potential

Coetzee

Vuuren

Stokstad

et al. 2012

Veterinary Microbiology

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A duplex real-time RT-PCR for the detection of bluetongue virus in bovine semen

Cited by 33 publications

References 26 publications

Longitudinal study of the detection of Bluetongue virus in bull semen and comparison of real-time polymerase chain reaction assays

Longitudinal study of the detection of Bluetongue virus in bull semen and comparison of real-time polymerase chain reaction assays

Bluetongue: Virus proteins and recent diagnostic approaches

Bluetongue virus genetic and phenotypic diversity: Towards identifying the molecular determinants that influence virulence and transmission potential

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