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Acanthamoeba is a genus of free-living amoebae that are among the most prevalent protozoa found in the environment. It is found worldwide and have been isolated from soil, air, and water. Human infections usually occur through exposure to different water resources, including freshwater, sea water, as well as domestic water supplies. Acanthamoeba keratitis (AK) is a severe sight-threatening ulceration of the cornea. The number of reported cases of AK is increasing worldwide every year due to increasing contact lens (CL) wear.Microscopic examination and culture have been used routinely for diagnosis of Acanthamoeba infections, but conventional and real-time polymerase chain reactions (PCR) assays have been developed more recently for sensitive and specific detection because rapid diagnosis is vital for successful therapy of AK.