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Subgenus classification of Acanthamoeba remains uncertain. Twenty-three reference strains of Acanthamoeba including 18 (neo)type-strains were subjected for classification at the subgenus level by riboprinting. PCR/RFLP analysis of 18S rRNA gene (rDNA). On the dendrogram reconstructed on the basis of riboprint analyses, two type-strains (A. astronyxis and A. tubiashi) of morphological group 1 diverged early from the other strains and were quite distinct from each other. Four type-strains of morphological group 3, A. culbertsoni, A. palestinensis, A. healyi were considered taxonomically valid, but A. pustulosa was regarded as an invalid synonym of A. palestinensis. Strains of morphological group 2 were classified into 6 subgroups. Among them, A. griffini which has an intron in its 18S rDNA was the most divergent from the remaining strains. Acanthamoeba castellanii Castellani, A. quina Vil3, A. lugdunensis L3a, A. polyphaga Jones, A. triangularis SH621, and A. castellanii Ma strains belonged to a subgroup, A. castellanii complex. However, A. quina and A. lugdunensis were regarded as synonyms of A. castellanii. The Chang strain could be regarded as A. hatchetti. Acanthamoeba mauritaniensis, A. divionensis, A. paradivionensis could be considered as synonyms of A. rhysodes. Neff strain was regarded as A. polyphaga rather than as A. castellanii. It is likely that riboprinting can be applied for rapid identification of Acanthamoeba isolated from the clinical specimens and environments.
Figures
Fig. 1 Agarose electrophoretic pattern of PCR products from Acanthamoeba 23 strains. Hind III digested λ phage DNA was used as DNA size marker(M).
Fig. 2 Schematic representation of riboprints of 23 strains of Acanthamoeba by 10 kinds of restriction enzymes. Hae III digested ΦΧ174 DNA was used as the size marker.
Fig. 3 Dendrogram of 23 strains of Acanthamoeba constructed by UPGMA method using Phylip ver. 3.5 based on genetic divergence estimates.
Tables
Table 1 List of 23 strains of Acanthamoeba analysed in this study
Table 2 Proportional homologous (values above the diagonal) and estimates of genetic divergence (values below the diagonal) among 23 strains of Acanthamoeba
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