The invention discloses a chicken embryo culture method of Portunus tritubereulatus reovirus and is characterized by comprising the following steps of: using SPF chicken embryo as an in vitro culture system of Portunus tritubereulatus reovirus, selecting healthy chicken embryo of 3 and 4 days-old, inoculating a prepared Portunus tritubereulatus reovirus inoculated solution onto a chorioallantoic membrane of the chicken embryo, continuously culturing until the chicken embryo is dead, collecting dead chicken embryo cultured for 3-10 days, separating the chorioallantoic membrane and amniotic fluid, mixing the chorioallantoic membrane and the amniotic fluid, homogenizing the mixture, centrifuging and filtering to obtain a supernatant of Portunus tritubereulatus reovirus. The invention has the following advantages: a more efficient and adaptive Portunus tritubereulatus reovirus in vitro culture method is provided for rapid and effective in vitro subcultring of the virus; and the method overcomes the defect that there is no adaptive culture system during the culture process of Portunus tritubereulatus reovirus, fills the blank of chicken embryo culture system of crab virus and has laid a foundation for further research and application.