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Copyright © 2015 J.K. Downes et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Peer-reviewed., Amoebic gill disease (AGD) is a proliferative gill disease of marine cultured Atlantic salmon Salmo salar, with the free-living protozoan Neoparamoeba perurans being the primary aetiological agent. The increased incidence of AGD in recent years presents a significant challenge to the Atlantic salmon farming industry in Europe. In this study, a real-time TaqMan® PCR assay was developed and validated to detect Neoparamoeba perurans on Atlantic salmon gills and further used to monitor disease progression on a marine Atlantic salmon farm in Ireland in conjunction with gross gill pathology and histopathology. The assay proved specific for N. perurans, with no cross-reactivity with the related species N. pemaquidensis, N. branchiphila or N. aestuarina, and was capable of detecting 2.68 copies of N. perurans DNA μl−1. Although the parasite was detected throughout the 18 mo period of this study, mortality peaks associated with clinical AGD were only recorded during the first 12 mo of the marine phase of the production cycle. The initial AGD outbreak resulted in peak mortality in Week 17, which was preceded by PCR detections from Week 13 onwards. Freshwater treatments were an effective method for controlling the disease, resulting in a reduction in the weekly mortality levels and also a reduction in the number of PCR-positive fish. In comparison to traditional diagnostic methods, our PCR assay proved to be highly sensitive and a valuable tool to monitor disease progression and, therefore, has the potential to provide information on the timing and effectiveness of treatments.