Abstract: A real-time quantitative RT-PCR assay was developed for detection of Swine influenza virus (SIV).Primers and probe were designed based on NP gene of SIV by Primer Express 2.0 software. The plasmid containing the target sequence was constructed to detect the sensitivity and prepare the standard curve. The real-time RT-PCR assay had a detection limit of 20 copies, with a dynamic range of detection between 2×10⁸ to 2×10² copies/μL. The standard curve was prepared based on the linear relationship between the amount of plasmid RNA and cycle threshold (Ct).The method is specific for SIV, which failed to react with the classical swine fever virus, pseudorabies virus, foot and mouth disease virus and porcine reproductive and respiratory syndrome virus nucleic acid.The real-time RT-PCR assay that described with high sensitivity, specificity and accuracy is considered to be a powerful tool for the rapid detection and quantification of SIV.

Key words: SIV; real-time quantitative RT-PCR; TaqMan probe