| Summary |
A quantitative one-step SYBR Green I-based reverse-transcriptase (RT)- PCR system was developed for the detection of bovine ephemeral fever virus in samples. The primer pair was designed based on conserved sequences in the G protein and evaluated for clinical diagnosis. A serial of ten-fold dilutions of the viral genome were used as templates for the reaction in which they served as standards to quantitate unknown viral samples. By using this system it was shown that as few as 300 copies of a viral genome could be detected. The diagnostic sensitivity of the real time RT-PCR and conventional RT- PCR, nested PCR and cell culture method in the detection of 72 clinical samples suspected to have BEFV infections were 19.04%, 12.8%（8/72）, 19.04%（14/72）and 18.06%（13/72）, respectively. The results indicated that the assay developed in this study was more sensitive than the conventional RT-PCR and isolation for the detection of BEFV. The novel assay is an excellent diagnostic tool for BEF infection.
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