Riboprinting ofNaegleria spp.: Small-subunit versus large-subunit rDNA

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Abstract
The nonpathogenic amoebaNaegleria lovaniensis is closely related to the human pathogenN. Fowleri. Both grow at a maximal temperature of 45°C and, therefore, are often found together in the environment. As they are morphologically inseparable at the light-microscope level, refined techniques are necessary to separate the two species. I have used restriction-fragment-length polymorphism analysis of the polymerase chain reaction (PCR)-amplified ribosomal RNA gene, or riboprinting, to distinguish between the differentNaegleria spp. Riboprints generated from the small subunit and the large subunit separateN. fowleri fromN. lovaniensis. To examine the taxonomic relationships among allNaegleria spp., analysis of the large subunit has to be performed; the small subunit contains a 1.3-kb group I intron, which interferes with tree building based upon restriction sites.