Abstract: Grass carp (Ctenopharyngodon idella) is an important freshwater economic fish in China and its production was 5.5×109 kg in 2014, accounting for 13% of global freshwater fishery production, which was the highest output and consumption of freshwater fish worldwide. Hemorrhagic disease caused by grass carp reovirus (GCRV) seriously affects the grass carp production. This study isolated and purified a new strain of GCRV from diseased grass carp from muscle of dead fish with typical symptoms of hemorrhagic diseases, whihc was further determined.by electron microscope and RT-PCR. A large number of virus particles with 70-80 nm in diameter were observed by electron microscope, which was diagnosed as genotype Ⅱ (GCRV-Ⅱ) by RT-PCR. The titer of virus was determined by absolute quantitative PCR (2.97×10³ copy/μL). The GCRV artificial infection experiments with 3 three different concentrations and one PBS control group were conducted with new method of lavage. The mortality rate of three experimental groups were all about 80%, while only one fish died in PBS control group. The typical symptoms of hemorrhagic disease were found in the abdomen, fin base and gill cover of dead fish. GCRV-Ⅱ was detected from four randomly selected dead fish using RT-PCR. These results indicate that lavage could be a new effective way for GCRV artificial infection experiment.