Ascaridia galli is a common nematode found in the intestine of domesticated chickens. The objectives of the study were to conduct a coprological and serological survey on the prevalence of ascaridiosis in laying hens of commercial farms. The farms recently adopted a breeding programme, where the hens have access to outdoor pens. Different amounts of Ascaridia eggs were detected in five of seven studied farms, while the other two farms were found to be free from the parasite. Serological tests revealed a seroprevalence of 21.8% (range 7.6-95%). No positive serum samples were detected in the same farms with previous negative coprological analysis. Western blot analyses confirmed the results obtained by the enzyme-linked immunosorbent assay (ELISA) test. In four experimentally infected hens, a progressive increase of the IgG antibody levels was observed, surpassing the cut-off point established for ELISA test 6 weeks post-infection. Serological tests are able to detect the infection before the eggs of the parasite appear in the faeces of infected hens, providing a useful tool to detect infections with Ascaridia spp. in avian farms.