Abstract: To establish the fluorescent realtime quantitative PCR (FQRTPCR) assay for rapid detection of egg drop syndrome virus, a pair of primers and a TaqMan probe were designed according to the sequences from highly conserved regions of the hexon protein gene of egg drop syndrome virus (EDSV). A positive recombinant plasmid was employed as standard template for the construction of standard curve. The detection limit of the assay was 10 copies DNA·μL-1 and the FQRTPCR was reproducible, as shown by satisfying its wide dynamic range from 101 to 108 copies DNA·μL-1. This assay is specific and has no crossreaction with DNA of other avian virus. The quantity of EDSV in eggs after artificial infection with NE4 strain of EDSV were detected by using the developed FQRTPCR method. Compared with common PCR assay, the FQRTPCR method is more sensitive and more suitable for detection of egg drop syndrome virus in vivo and in vitro.