Dr. Murai Éva - Gubányi András szerk.: Parasitologia Hungarica 28. (Budapest, 1995)

© Hungarian Natural History Museum Hungarian Society of Parasitologists Parasit, hung., 28: 5-12, 1995 Isolation and restriction enzyme analysis of mitochondrial DNA (mtDNA) of Hungarian Acanthamoeba isolates from human eye infection and from the environment Zsuzsanna SZÉNÁSI

1

, Takuro ENDO

2

, Kenji VAGUA

2

, Edit URBAN

1

, Mihály VÉGH

3 and Erzsébet NAGY

1 1 Department of Clinical Microbiology, Albert Szent-Györgyi Medical University, H-670J Szeged, P.O. Box 482, Hungary 2 Department of Parasitology, National Institute of Health, Kami-Osaki, Shinagawa-Ku, Tokyo 141, Japan ^Department of Ophthalmology, Albert Szent- Györgyi Medical University, H-6720 Szeged, Korányi fasor 7-11, Hungary (Received 18 June, 1995) Abstract: Restriction fragment length polymorphism (RFLP) analysis was used for the typing of Acanthamoeba isolates from the contact lens container and corneal scrapings of a patient with keratitis (CI) and for typing isolates from the environment (Dun, Mos). The Acanthamoebae were isolated on agar plates covered with a suspension of Enterobacter aerogenes. The isolates were cloned by micromanipulation, then cultured and maintained on agar plates spread with a layer of heat-treated Escherichia coli DH

1. The cloned isolates were adapted and cultured axenically. The mitochondrial DNA (mtDNA) was prepared by alkaline lysis and digested by Bg\ II and £coR I endonucleases. The three isolates have distinct RFLP phenotypes. On the basis of the fragment sizes obtained with two restriction enzymes, the mtDNA genotypes of the three Hungarian isolates are comparable to those of human isolates from different countries. As all strains of Acanthamoeba may have pathogenic potential, the mtDNA RFLP phenotype may be a useful aid in the definition of their pathogenic potential. Key words: Acanthamoeba, eye infection, RFLP analysis, mtDNA INTRODUCTION Infections caused by small free-living amoebae are generally not very well known or recognized clinically. Most laboratories are unfamiliar with the diagnostic methods of such infections and, therefore, do not use them on a routine basis. Free-living Acanthamoeba spp are common predators of bacteria. They are ubiquitous in nature. Dry cysts can survive