Molecular detection and genotype identification of Acanthamoeba species from bronchoalveolar lavage of patients with pulmonary symptoms suspected for cancer

Abstract Acanthamoeba spp. are the most common free-living amoeba worldwide, which cause life-threatening diseases such as Granulomatous Amoebic Encephalitis, pulmonary infection, and amoebic keratitis. This study is aimed to identify FLA and Acanthamoeba genotypes among patients with pulmonary symptoms suspected for cancer in Kashan hospitals, center of Iran.This cross-sectional study was conducted on 97 bronchoalveolar lavage samples of patients with symptoms of respiratory problems suspected of lung cancer admitted to Shahid Beheshti hospital of Kashan 2019 to 2020. The samples were cultured onto 1.5% Non-nutrient agar enriched with killed Escherichia coli and examined for the presence of FLA. After isolation of amoeba and DNA extraction, Acanthamoeba spp. were determined by Polymerase Chain Reaction using JDP1 and JDP2 primers, which amplified a 490 bp fragment from the 18 S rDNA gene. Eighteen Acanthamoeba isolates were sequenced and genotypes were identified. The prevalence of FLA and Acanthamoeba and the relation between symptoms and demographic variables were analyzed by the SPSS.16. The prevalence of FLA and Acanthamoeba in BAL samples was 86.6% and 73.2%, respectively. All Acanthamoeba isolates were belonged to the T4 genotype. The most symptoms among Acanthamoeba positive patients were dyspnea, cough, but the difference was not statistically significant. The results have shown a high prevalence rate of FLA and Acanthamoeba in BAL of patient suspected to cancer in Kashan. Since the T4 is a pathogenic genotype of Acanthamoeba, health training and improving sanitation level help to prevent infection.