Objective HCV will be inoculated rapidly enough to tree shrew liver in vivo by using abdominal transverse operation and hepatic portal vein injection , to improve the rate of HCV infection in tree shrews. Methods Tree shrew peritoneal were opened by the transverse abdominal operation, the hepatic portal vein were found, HCV were inoculated by hepatic portal vein injection. Results 20 tree shrews are transverse operation and hepatic portal vein injection，The success rate of operation and hepatic portal vein injection accuracy rate reached 100%, HCV is completely into the tree shrew liver. Tree shrews had good health and survived after the postoperative nursing care, the operation had no effect on the blood sampling and testing. The other 2 tree shrews of abdominal longitudinal incision were not inoculated by the hepatic portal vein injection, because hepatic portal vein was covered by the liver and was not easy to be found . Conclusion Abdominal transverse has many advantages , including fully exposed the hepatic portal vein, convenient operation, in accordance with physiological solution planing, less tissue trauma,, good wound healing and so on, it is suitable for hepatic portal vein injection, which lays a foundation for the further study of HCV infection of tree shrew, and has certain experience and reference value for other related experimental animal and animal experiment.