Objective: Infectious laryngotracheitis (ILT) is an economically important viral respiratory disease of poultry around the world. The aim of this study is investigate the presence of anti-ILTV antibodies in commercial layers and village chickens in the Central Anatolia Region by indirek ELISA. Material and Methods: A total of 878 blood samples were collected from 28 commercial layer flocks (n=733) comprising 17 unvaccinated (n=268), 11 vaccinated (n=465), and village chickens (n=145) at 7 different localisation in the Central Anatolia Region. Sera were obtained from the collected blood samples. Anti-ILT virus (ILTV) antibodies were determined by commercial indirect ELISA kit. Results: Anti-ILTV antibodies were determined of the investigated 145 unvaccinated village chickens, 268 unvaccinated commercial layer chickens, and 465 vaccinated commercial layer chickens, at ratios of 86.90% (n=126), 90.67% (n=243), and 35.70% (n=166), respectively. Although 5 commercial layer flocks were vaccinated, no antibody response was detected. Conclusion: It was determined that ILT infection poses a serious risk especially in unvaccinated commercial layer flocks and village chickens in the Central Anatolia Region. We believe that it would be a more correct approach to conduct phylogenetic analyzes of strains isolated from ILT outbreaks and to focus on the source of infection in further studies, with the prediction of vaccine-induced spillage due to the proximity of the flocks.