Abstract: Objective To understand the distribution of avian influenza A (H7N9) virus in human and poultry in Lishui, Zhejiang province and provide evidence for the prevention and control of human infection with avian influenza A (H7N9) virus. Methods Fluorescent Quantitative PCR was conducted to detect the samples from the human cases with fever and respiratory symptoms in medical institutions and environmental samples from April 2013 to February 2014 in Lishui. Results A total of 768 cases with mild influenza like symptoms and 605 cases with severe respiratory tract infection were detected, however, all the cases were H7N9 virus negative, and 92 cases with pneumonia with unknown origin were detected, only 1 was positive for H7N9 virus (1.09%). A total of 1317 poultry stool samples, smears of cages and egg shell surface, nasal and throat swabs of poultry and samples from unexplained dead poultry/wildfowl were detected, and only 22 (1.67%) were H7N9 virus positive, all the positive samples were from 8 live poultry markets in 4 counties (district). Conclusion It is the first time to detect avian influenza A (H7N9) virus in some live poultry markets in Lishui, indicating that the avian influenza A (H7N9) virus transmission among the poultry has occurred. The risk of human infection exists.