Diagnostic procedures for detection of infectious laryngotracheitis virus in tracheas of experimentally infected chickens, including the indirect fluorescent antibody test (IFAT), immunoperoxidase (IF), virus isolation (VI), histopathology, polymerase chain reaction (PCR), and DNA hybridization, were performed and compared. Using VI as a reference, we calculated the sensitivity and specificity of the tests. The sensitivities of IF, IFAT, histopathology, PCR, and hybridization were 100%, 93%, 7%, 27%, and 0%, respectively, and the specificities of IP, IFAT, histopathology, PCR, and hybridization were 93%, 93%, 100%, 100%, and 100%, respectively. Histopathology, PCR, and hybridization were more specific bur lacked sensitivity compared to IP and IFAT. IP and IFAT were equally specific, bur IP was more sensitive than IFAT. Based on these results, IP performed better than any other test.