Providing suitable drinking water for human use is a major goal in all countries, and contamination of drinking water with microorganisms represents a great danger to human health. Free-living amoebae (FLAs, Pathogenic) are found in all natural and manufactured environments, including aquatic environments. Several FLAs such as Acanthamoeba spp. infect humans, causing dangerous diseases. The purpose of this investigation was to search about FLAs in tap water in Nasr City, Cairo.  FLAs were detected in 59 (53.63 %) out of 110 tap water samples which were morphologically identified as two Acanthamoeba spp. The molecular identification to the species level was carried out, using a simple PCR technique and DNA sequencing.  Acanthamoeba spp. were recorded in the gene bank under accession numbers Acanthamoeba MT944118 and Acanthamoeba MT944119.