Evaluation of Acanthamoeba Species from Tehran Surface Water Sources Using Real Time PCR Method

Abstract

Acanthamoeba spp. is the ubiquitous potentially pathogenic free-living amoebae in nature such as water sources. Recently Acanthamoeba keratitis (Ak) continue to rise in Iran. Most of patients report a history of contact with water sources before the onset of disease. The main aim of the present study was to determine the occurrence of Acanthamoeba spp. in the recreational water sources of Tehran, Iran using morphological and molecular based tests. A total of 70 surface water samples were collected from environmental sources, including parks pools and swimming pools. The samples were filtrated and transferred to non-nutrient agar plates seeded with Escherichia coli and incubated for 2 to 7 days at 30°C or 42°C. The plates were examined by microscopy to morphologically identify Acanthamoeba spp. Following DNA extraction, PCR was used to confirm the microscopically identification. A total of 11 out of 35 samples of parks pools and 3 out of 35 of swimming pools were positive for Acanthamoeba species based on the morphological criteria, and all were confirmed by PCR method. The significant frequency of Acanthamoeba spp in surface water sources of Tehran is of importance to beconsidered from public health point of view by authorities.