Abstract: Goose astrovirus (GAstV), as an emerging pathogen causing visceral gout and high mortality in goslings and result in huge economic losses. Thus, it is very important to develop credible detection methods to prevent this diseases wide spread in the absence of effective treatment measure. In order to establish a rapid, specific and sensitive method for detection of genotype GAstV Ⅰ and GAstV Ⅱ viruses, a duplex TaqMan real-time quantitative reverse transcription PCR (RT-qPCR) assay was established. Two pairs of specific primers and two kinds of TaqMan probes labeled with different fluorescent groups according to the RdRp gene sequences were designed and synthesized. The results showed that the minimum detectable amounts of GAstV Ⅰ and GAstV Ⅱ were 43.3 and 6.49 DNA copies·μL^-1, respectively; the coefficient of variation of repeated tests was less than 0.5%; and the method had no cross reaction to viral nucleic acids such as DPV, GPV, GTMUV, GRV, AIV (H9N2), etc. In summary, the results above indicate that this duplex real-time PCR method can distinguish GAstV Ⅰ and GAstV Ⅱ with high sensitivity and specificity. It can be used for the clinical rapid differential diagnosis of GAstV Ⅰ and GAstV Ⅱ, and quantitative analysis of two genotypes of GAstV.

Key words: GAstV Ⅰ, GAstV Ⅱ, double real-time RT-PCR, TaqMan probe