Van Eeden, CharmaineZaayman, DewaldVenter, Marietjie2015-07-012015-07-012014-01Van Eeden, C, Zaayman, D & Venter, M 2014, 'A sensitive nested real-time RT-PCR for the detection of Shuni virus', Journal of Virological Methods, vol. 195, no. 1, pp. 100-105.0166-093410.1016/j.jviromet.2013.10.008http://hdl.handle.net/2263/45825Recently Shuni virus (SHUV) has been identified in clinical cases of neurological disease in horses in South Africa. Being that it was one of the less recognized orthobunyaviruses, with limited clinical descriptions of disease dating back to the 1960s and 1970s, SHUV-specific assays were never developed. In this study, the development of a nested real-time PCR assay is described for the detection of SHUV by means of melt-curve analysis using fluorescence resonance energy transfer (FRET) probe technology. The assay was validated against previously positive clinical specimens and a dilution series of controls. This assay was applied in the screening of 386 clinical specimens of horses with neurological signs. SHUV could be amplified from cell culture, blood and tissue specimens from horses with acute disease. This is a rapid and sensitive assay that may be implemented in both diagnostic and research laboratories.en© 2013 Elsevier B.V. All rights reserved. Notice : this is the author’s version of a work that was accepted for publication in Journal of Virological Methods. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Journal of Virological Methods, vol. 195, no. 1, pp. 100-105, 2014. doi : 10.1016/j.jviromet.2013.10.008Real-time RT-PCRFRET probesDiagnostic PCRShuni virus (SHUV)fluorescence resonance energy transfer (FRET)A sensitive nested real-time RT-PCR for the detection of Shuni virusPostprint Article