Molecular characterization of Acanthamoeba strains isolated from the oral cavity of hemodialysis patients in Iran.

Free-living amoebae (FLA) of the genus Acanthamoeba are opportunistic pathogenic agents able to cause life-threatening infections in immunosuppressed patients. Chronic kidney disease impairs adaptive and innate immunity. Thus, patients with chronic kidney disease are prone to opportunistic infections by potentially pathogenic FLA. Therefore, in the present study, the investigation of Acanthamoeba genotypes isolated from the oral cavity of hemodialysis patients of reference hospitals in Iran was aimed, using both morphology and molecular (sequence-based analysis) tools. Furthermore, classification of the strains at the genotype level was performed on the basis of differences in the diagnostic fraction 3 (DF3) region of the 18S rRNA gene. The pathogenic potential of the isolated amoebae was also determined using thermotolerance and osmotolerance assays. Out of the 187 oral cavity samples investigated, nine (4.8%) were positive for FLA. DNA sequencing of the ASA.A1 region of the 18S rRNA gene revealed that the isolated strains belonged to the Acanthamoeba T1 and T4 genotypes. Genotype T1 was isolated for the first time from a patient in Iran. Interestingly, the T1 strain (AN2 strain) exhibits a high pathogenic potential in tolerance assays. The pathogenicity assay revealed that five strains were able to grow at high temperatures (37-40 °C) and high osmolarity (0.5 and 1 M D-mannitol) conditions; thus, they were considered as potentially pathogenic strains. Moreover, two of the patients were positive for Vermamoeba genus. The present study is the first report of genotype T1 isolation in Iran and the first to identify the occurrence of Acanthamoeba and Vermamoeba genera in patients undergoing hemodialysis worldwide. Monitoring hemodialysis and renal failure patients should be a priority for possible control of Acanthamoeba and other FLA-related diseases.