In order to understand the infection status and main epidemic subgroups of avian leukosis virus (ALV) in indigenous chicken of Hubei province,7 477 samples were collected from three local core breeds groups:the Macheng green-shell layer chicken,the Jianghan chicken and the Jingyang chicken groups,followed by the detection of the egg white p27 antigen,isolation and identification of ALV from roosters,and sequence analysis of the gp85 gene. The results showed that the positive rate of p27 antigen in the three breed groups were 1.32%,6.54% and 2.82%,respectively,and the isolation rates of ALV from roosters were 6.11%,30.14% and 8.64%,respectively,indicating that the ALV infection rate in chickens in Hubei province was relatively serious. The 15 ALV isolates can be divided into two subsets using PCR. Genetic evolution analysis of the gp85 gene showed that the J subgroup (ALV-J) and K subgroup (ALV-K) were prevalent in the three indigenous chicken breed groups,and co-infection of ALV-J and ALV-K was also found in all the three groups. The results indicated that the ALV infection in indigenous chickens in Hubei province was complicated.