A sensitive nested real-time RT-PCR for the detection of Shuni virus

Abstract

Recently 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.