title: 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.
authors.affiliation: Zoonoses Research Unit, Department of Medical Virology, Faculty of Health Sciences, University of Pretoria, South Africa. Electronic address: charmaine.vaneeden@gmail.com.
authors.forename: C
authors.initials: C
authors.lastname: Van Eeden
authors.forename: D
authors.initials: D
authors.lastname: Zaayman
authors.forename: M
authors.initials: M
authors.lastname: Venter
mesh_terms: D000818:Animals; D002044:Bunyaviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D031541:Fluorescence Resonance Energy Transfer; D006734:Horse Diseases / Q000175:diagnosis* / Q000821:virology; D006736:Horses; D025202:Molecular Diagnostic Techniques / Q000379:methods; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D016851:Orthobunyavirus / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013019:South Africa; D013997:Time Factors; D014730:Veterinary Medicine / Q000379:methods*
chemical_list: D015345:Oligonucleotide Probes; D012367:RNA, Viral
keywords: Diagnostic PCR; FRET probes; Real-time RT-PCR; Shuni virus