The significant economical consequences of infections with Ascaris suum in pigs are already well documented. However, due to the subclinical nature of the disease and the lack of practical diagnostic means, ascariasis often remains undiagnosed. Here we describe the development and evaluation of a novel indirect ELISA using the purified A. suum haemoglobin (AsHb) molecule as an antigen. Initial validation using sera from 190 pigs experimentally infected twice a week with A. suum and Trichuris suis (25 and 5 eggs kg(-1)day(-1) respectively) demonstrated that the AsHb ELISA is able to detect long-term exposure to A. suum with a high sensitivity and specificity (99.5% and 100.0% respectively). Furthermore, this serological technique proved to be more sensitive than faecal examination on week 7 and 14 of the experiment (99.5% and 100% compared to 59.5% and 68.4% respectively). Cross-reactivity caused by T. suis infection was shown to be limited after analysing sera from pigs with an experimental T. suis mono-infection. Seroconversion was shown to occur from week 6 onwards in pigs receiving 100 A. suum eggs 5 times a week. Preliminary testing of the ELISA on six randomly selected farms confirmed the results obtained in the artificial infection trials, showing a higher sensitivity of the serologic method compared to faecal examination. Finally, the ELISA was used to investigate Ascaris infection rates on 101 conventional Flemish pig farms. The results showed that on 38.6% of the farms less than 20% of the tested samples were seropositive, while in 19.8% of the farms 80-100% of all pigs were seropositive. The results of this study suggest that the AsHb ELISA could provide pig farmers and veterinarians with an easier and more sensitive way to estimate the overall prevalence of A. suum on their farm.