Development of a double antibody sandwich ELISA method for the detection of avian leukosis virus

A double antibody sandwich ELISA(DAS-ELISA) method was developed with monoclonal antibody against p27 of subgroup J avian leukosis virus(ALV) HB09JY03 as capture antibody and polyclonal antibody against the p27 as detector antibody to detect ALV rapidly and effectively.The sensitivity limit of the method was 1.25 ng/mL.No cross-reaction was observed with avian infectious bronchitis virus,Marek's disease virus,avian infectious laryngotracheitis virus and so on.It was suitable for detection of virus from the cloacal swabs of experimentally infected chicken at 12 day post-infection.A total of 491 albumin samples and 180 cloacal swabs were tested using the DAS-ELISA,and 96.5% and 93.8% agreement were found with the results of PCR,respectively.All of these results showed that the DAS-ELISA was a convenient,rapid and sensitive method for the detection of ALV,and the present study provided a tool for the early diagnosis of ALV and clarification of ALV in breeder chickens.