Abstract: To establish a real-time fluorogenetic quantitative RT-PCR assay for detection of classical swine fever virus (CSFV)，the 5′ non-translated region sequences of CSFV in GenBank were aligned and a pair of specific primers was designed from the conserved sequence within 5′ non-translated region. The reactive conditions were optimized to improve the sensitivity and specificity of the assay. The results of reproducibility of this assay were reliable and the intra assay variations were not significant. The specificity test proved that this assay had a high specificity which could not detected PRRSV，PRV and PCV2. The assay also proven to be specific，and the detection limit was up to 2×102 copies. The accuracy of real-time fluorogenetic quantitative RT-PCR was evaluated by testing 15 clinical samples. The results of real-time fluorogenetic quantitative RT-PCR were consistent with results of rabbit-cross reaction test，which suggested that real-time quantitative RT-PCR assay may be used as a powerful tool for repaid detection of CSFV.

Key words: classical swine fever virus; real-time fluorogenetic quantitative RT-PCR〖