In order to develop an appropriate method for high-throughput detection of avian infectious laryngotracheitis（AILT），a real-time PCR assay was established with a pair of specific primers and a probe based on the conservative zone of gB gene sequence of AILT，then its specificity and sensitivity were evaluated，and its clinical application was tested. The results showed that the selected primers and probe failed to cross-react with other common poultry viruses，with the detection limit of 100 copies/reaction. 17 out of 480 clinical samples collected from 13 farms were positive. The results completely conformed to the test result by common PCR assay. As a conclusion，AILT could be rapidly detected by the established method with high specificity，high sensitivity and less time-consuming.