BackgroundA coproantigen enzyme-linked immunosorbent assay (ELISA) has recently been proposed for detecting ascarid infections in chickens. The excretion pattern of ascarid antigens through chicken faeces and the consistency of measurements over the course of infections are currently unknown. This study evaluates the pattern and repeatability of worm antigen per gram of faeces (APG) and compares the diagnostic performance of the coproantigen ELISA with a plasma and egg yolk antibody ELISA and McMaster faecal egg counts (M-FEC) at different weeks post-infection (wpi).MethodsFaecal, blood and egg yolk samples were collected from laying hens that were orally infected with a mix ofAscaridia galliandHeterakis gallinarumeggs (N= 108) or kept as uninfected controls (N= 71). Measurements including (a) APG using a coproantigen ELISA, (b) eggs per gram of faeces (EPG) using the McMaster technique and (c) ascarid-specific IgY in plasma and in egg yolks using an ascarid-specific antibody ELISA) were performed between wpi 2 and 18.ResultsTime-dependent significant differences in APG between infected and non-infected laying hens were quantified. At wpi 2 (t(164)= 0.66,P= 1.00) and 4 (t(164)= −3.09,P= 0.094) no significant differences were observed between the groups, whereas infected hens had significantly higher levels of APG than controls by wpi 6 (t(164)=  −6.74,P< 0.001). As indicated by a high overall repeatability estimate of 0.91 (CI = 0.89–0.93), APG could be measured consistently from the same individual. Compared to McMaster and antibody ELISA, coproantigen ELISA showed the highest overall diagnostic performance (area under curve, AUC = 0.93), although the differences were time-dependent. From wpi 6 to 18 coproantigen ELISA had an AUC > 0.95, while plasma IgY ELISA showed the highest diagnostic performance in wpi 2 (AUC = 0.95). M-FEC had the highest correlation with total worm burden, while APG had highest correlations with weights and lengths ofA. galli.ConclusionAscarid antigen excretion through chicken faeces can be measured with high accuracy and repeatability using a coproantigen ELISA. The antigen excretion increases over time, and is associated with worm maturation, particularly with the size ofA. galli. Our results suggest the necessity of complementary use of different diagnostic tools for a more accurate diagnosis of infections.Graphical AbstractSupplementary InformationThe online version contains supplementary material available at 10.1186/s13071-023-05782-5.