Identification of Acanthamoeba at the Generic and Specific Levels Using the Polymerase Chain Reaction

MICHAEL H. VODKIN, DANIEL K. HOWE, GOVINDA S. VISVESVARA, GERALD L. McLAUGHLIN

Research output: Contribution to journalArticlepeer-review

67 Scopus citations

Abstract

ABSTRACT. We have adapted the polymerase chain reaction to identify strains of Acanthamoeba. Using computer‐assisted analysis, primers were designed from an anonymous repetitive sequence and from published sequences of 18S and 5S ribosomal RNA genes of A. castellanii. Amplification of a short ribosomal DNA target (272 base pairs) at restrictive annealing conditions (>50° C) resulted in a single band that was unique for the genus and distinguished Acanthamoeba from Naegleria. This assay functioned with fresh and formalin‐fixed cells as starting material. Amplification of longer targets (400–700 base pairs) at less restrictive annealing conditions (<47° C) led to more than one band. This multiple banding pattern could reproducibly classify Acanthamoeba at the strain level and was, in certain cases, diagnostic for known pathogenic strains. However, these assays need to be further refined to make them relevant for clinical purposes.

Original languageEnglish
Pages (from-to)378-385
Number of pages8
JournalJournal of Eukaryotic Microbiology
Volume39
Issue number3
DOIs
StatePublished - May 1992

Keywords

  • Amoeba
  • Naegleria
  • human pathogen
  • repetitive DNA
  • ribosomal DNA

ASJC Scopus subject areas

  • Parasitology

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