Abstract: A pair of primers were designed for detection of avian influenza viruses(AIV). 4 isolates belonging to different subtypes were used to infect MDCK cells, then total RNA were extracted from supernatant fluids after infecting 6，12，24，48，72 h. Throat, tracheal swabs samples from chicken were extracted too.SYBR Green I RT-PCR of M gene was done with Rotor Gene 3000 PCR amplifier. We attempted to establish a SYBR Green I RTPCR method to detect AIV.At the same time, the results of SYBR Green I RT-PCR were compared with those of conventional RT-PCR and virus titration.The results showed that the assay was sensitive, specific and rapid enough to detect AIV RNA genome in cell culture supernatant and swabs samples, while other viruses were not detected. As such, the described method is reliable and provides faster detection of AIV than other conventional methods.