Species Identification of Acanthamoeba Strains Isolated from Patients Referring to Farabi Eye Reference Center Using PCR-RFLP Method

Introduction: Acanthamoeba is an opportunistic protist, which is ubiquitously distributed in the environment. Infection with Acanthamoeba spp. poses threat to human health, such as, Acanthamoeba keratitis (AK) that is a vision-threatening infection of the cornea. This study aimed to identify the species of Acanthamoeba strains isolated from cornea of keratitis patients by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Methods: Acanthamoeba isolates investigated in this cross-sectional study, were collected from patients referring to Farabi Eye Reference Center. All ۱۰ isolates were subjected to species identification using DNA based method. BspLI (NlaIV) and HpyCH۴IV restriction enzymes were used to categorize the PCR amplified DNA by PCR-RFLP method. Results: Six samples were identified as Acanthamoeba palestinensis and ۴ isolates as Acanthamoeba culbertsoni, which implies that all the isolates belong to pathogenic strains of Acanthamoeba. Conclusion: Acanthamoeba can enter the corneal tissue and survive in the eye, which results in AK. To the authors' knowledge, no study is available on species identification of this genus using these enzymes and technique. This is the first time in Iran that Acanthamoeba isolates are subjected to species identification using PCR-RFLP method.

Keywords:

Acanthamoeba Keratitis , Cornea , DNA Restriction Enzymes , PCR-RFLP