Espach, AnelRomito, M.Viljoen, G.J.Boomker, Jacob Diederik Frederik2012-12-122012-12-1220122002Espach, A, Romito, M, Nel, LH & Viljoen, GJ 2002, 'Development of a diagnostic one-tube RT-PCR for the detection of Rift Valley fever virus’. Onderstepoort Journal of Veterinary Research, vol. 69, no. 3, pp. 247-252.0030-2465http://hdl.handle.net/2263/20695The articles have been scanned in colour with a HP Scanjet 5590; 600dpi. Adobe Acrobat v.9 was used to OCR the text and also for the merging and conversion to the final presentation PDF-format.Diagnosis of Rift Valley fever (RVF) is based on serology and virus isolation. The disadvantages of the former include poor sensitivity, high cost, risks associated with using infectious virus as antigen, the lengthy duration of ELISA as well as cross-reactivity with other Phleboviruses. We developed, optimised and evaluated a one-tube reverse-transcription-polymerase chain reaction (RT-PCR) for the detection of Rift Valley fever virus (RVFV) in ruminants. The PCR primers for this assay were designed to anneal to a region within the M segment of the virus genome, encoding glycoproteins G1 and G2. A PCR amplicon of 363 bp was obtained. The sensitivity of the assay was determined to be 0.25 TCID₅₀. This test should allow for the early and rapid detection of RVFV in both serum and whole blood. In addition, it could facilitate the quantification of antigen for the manufacture of current vaccines.© ARC-Onderstepoort and Faculty of Veterinary Science, University of Pretoria (original). © University of Pretoria. Dept of Library Services (digital).Veterinary medicineDiagnosisM segmentOne-tube RT-PCRRift Valley feverVeterinary medicine -- South AfricaArticle