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Identification of two genetic markers that distinguish pathogenic and nonpathogenic strains of Acanthamoeba spp.
Authors:Daniel K. Howe  Michael H. Vodkin  Robert J. Novak  Govinda Visvesvara  G. L. McLaughlin
Affiliation:Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, MO 63110‐1093, USA, US
Illinois Natural History Survey, Center for Economic Entomology, Champaign, IL 61820, USA, US
Division of Parasitic Diseases, Center for Infectious Diseases, Centers for Disease Control, Atlanta, GA 30333, USA, US
Department of Pathology and Laboratory Medicine, MS A128, Indiana University School of Medicine, 635 Barnhill Drive, Indianapolis, IN 46202‐5120, USA Fax: 317‐278‐2018; e-Mail: gmclaugh@indyvax.iupui.edu, IN
Abstract:Species-level identification of Acanthamoeba isolates is difficult and gives little or no indication of the isolate's pathogenicity. We identified two amplification-based genetic markers that were highly correlated with pathogenicity in Acanthamoeba spp. One marker, designed to amplify a 485-bp fragment of the small-subunit ribosomal RNA gene (ssrDNA), was preferentially amplified from the nonpathogenic strains; amplifications from the pathogenic strains yielded anomalous fragments of 650 and 900?bp. A second marker was developed on the basis of the anomalous 650-bp fragment. Primers to this sequence preferentially amplified a noncoding locus (called Ac6) only from the pathogenic strains. These two genetic markers may be useful for identification of pathogenic Acanthamoeba spp. strains.
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