Despite effective chemotherapy, fascioliasis remains a major public health problem in developing countries, with at least 17 million active infections, resulting in significant morbidity, late infection detection, and rapid reinfection after treatment. Therefore, alternative control strategies are mandatory. Proteases, myofibrillar proteins, are found only in invertebrates. In the present study, adult fresh Fasciola gigantica (F. gigantica) worms were homogenized; an antigen was purified and used to raise rabbit polyclonal antibodies (pAb). The purified pAb was then used in sandwich ELISA to detect Fasciola antigens in sera samples from a total 135 cattle.