In order to rapidly detect avian leukemia virus（ALV）and simultaneously identify its subgroup A（ALV-A）from J（ALV-J），specific primers and TaqMan fluorescent probes were designed based on different subgroup genes，followed by optimizing reaction conditions，a triple PCR for ALV universal type，subgroups A and J as well as a double fluorescent probe method for subgroups A and J were established. The results showed that the triple PCR could be used to specifically amplify all subgroups including A and J，with a sensitivity of 1×103 copies/μL；the double probe method，with strong specificity，could be used to detect the minimum copy concentration of 40 copies/μL. The established triple PCR was highly consistent（Kappa coefficient＞0.75）with the commercial real-time fluorescence PCR kit；for 24 semen samples from infected chickens，the results by the double probe method were the same as those by commercial real-time fluorescence PCR kit；subgroups A and J could be simultaneously identified by the two methods established in the paper. 394 clinical samples collected in Anhui Province were detected by the established methods，it was found that the total positive rate of ALV infection was 38.58%，mainly due to single infection with subgroup J，accompanied by single infection with subgroup A and mixed infection with subgroup A/J. In conclusion，the two established methods，with strong specificity，rapidity and application potential for clinical samples，could technically support rapid diagnosis and effective prevention and control of ALV in clinical practice as well as identification of single or mixed infection with subgroup A/J.