Background and rationale: Fascioliasis is a neglected parasitic infection that affects both humans and animals. It is caused by the trematode parasites known as Fasciola hepatica and Fasciola gigantica, which primarily infect the bile duct. Fascioliasis has a significant negative social and economic impact, with estimated infected rate in livestock (18%) and 2.6 million persons worldwide. The highly prevalence was recorded in least developed countries.  The actual challenge is to obtain a cheap and efficient diagnosis technique to diagnose the early mild stage of fascioliasis, parasitological examination techniques are usually effective in the final stage of infection when a large number of fascioliasis eggs are execrated in stool.  Aim: The aim of this study was conducted to develop and compare a novel diagnostic lateral flow assay and sandwich ELISA for detection of active fascioliasis infection. The conjugation of fascioliasis tegumental antibodies to AuNPs through covalent conjugation using EDC/Sulfo-NHS, to application in the setup of immuno-chromatography lateral flow assay (LFA) and sandwich ELISA. The novel LFA succeeded to diagnose (37 out of 38 true positive infected fascioliasis group), (2 out of 81 true negative other parasites infected group) and (0 out of 27 negative control group), the sensitivity, specificity, NPV and PPV of were 97.4%, 98.2%, 99.1%, and 94.8%, respectively. while the sensitivity, specificity, NPV and PPV of sandwich ELISA were 94.7%, 96.3%, 98.1%, and 90%, respectively. Conclusion: The higher sensitivity and specificity were recorded in LFA.  Lateral flow assays (LFA) technique could be used as Ideal diagnostic techniques in monitoring and control of fasciolosis infection.