Abstract
In recent years, real-time reverse transcription polymerase chain reaction (rRT-PCR) has become one of the most widely used methods for the diagnosis of infectious pathogens. The combined properties of high sensitivity, specificity, and speed, along with a low contamination risk, have made real-time PCR technology a highly attractive alternative to more conventional diagnostic methods. Numerous robust rRT-PCR systems have been developed and validated for important epizootic diseases of livestock, and in this chapter we describe an rRT-PCR protocol for the detection of bluetongue virus. The assay uses oligonucleotide primers to specifically amplify target regions of the viral genome and a dual-labeled fluorogenic (TaqMan®) probe which allows for the assay to be performed in a closed-tube format, thus minimizing the potential for cross-contamination.
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Batten, C., Frost, L., Oura, C. (2015). Real-Time Reverse Transcriptase PCR for the Detection of Bluetongue Virus. In: Cunha, M., Inácio, J. (eds) Veterinary Infection Biology: Molecular Diagnostics and High-Throughput Strategies. Methods in Molecular Biology, vol 1247. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2004-4_8
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DOI: https://doi.org/10.1007/978-1-4939-2004-4_8
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2003-7
Online ISBN: 978-1-4939-2004-4
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