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Digestion of 18s-srDNA with four enzyme, Dde I, Hae Ⅲ, Msp I and Rsa I, resulted in various DNA fragments. Using the method of Nei and Li(1979) for RFLP analysis, genetic divergence was not observed between A. culbertsoni and Acanthamoeba sp. YM-4. Between A. culbertsoni and Acanthamoeba sp. YM-5, genetic distance was 0.072, 0.365 between Acanthamoeba sp. YM-4 and A. polyphaga, and 0.278 between Acanthamoeba sp. YM-7 and A. polyphaga. In comparision with A. culbertsoni and A. polgphaga, Acanthamoebasp. YM-7 showed genetic distance of 0.335 and 0.153, respectively. Thus, Acanthamoeba sp. YM-7 is a similar species to A. polyphaga. Acanthamoeba sp. YM-5 is a different species from A. polyphaga, but closely-related to A. culbertsoni. Acanthamoeba sp. YM-4 can be classified as a subspecies or a strain of A. culbertsoni.
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