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Korean J Parasitol > Volume 33(4):1995 > Article

Original Article
Korean J Parasitol. 1995 Dec;33(4):341-348. English.
Published online Dec 20, 1995.  http://dx.doi.org/10.3347/kjp.1995.33.4.341
Copyright © 1995 by The Korean Society for Parasitology
Genetic status of Acanthamoeba spp. Korean isolates on the basis of RAPD markers
Yong-Pyo Hong,*1Seung-Hwan Oh,2Mi-Sook La,1 and Kyung-il Im1
1Department of Parasitology and Institute of Tropical Medicine, Yonsei University College of Medicine, Seoul 120-752, Korea.
2Department of Pediatrics, Yonsei University College of Medicine, Seoul 120-752, Korea.
Received August 07, 1995; Accepted November 10, 1995.

Abstract

Genetic status of Acanthamoeba spp. were tested on the basis of random amplified polymorphic DNA (RAPD) marker analysis. Four previously established Acanthamoeba species, 4 Korean isolates of Acanthamoeba sp., and one American isolate of Acanthamoeba sp. were analyzed by RAPD- PCR using an arbitrary decamer primers. Amplification products were fractionated by agarose gel electrophoresis and stained by ethidium bromide. Eighteen primers produced DNA amplification profiles revealing clear differences among 4 species. Nine of them also produced DNA amplification profiles which included some isolate-specific amplification products. On the basis of amplified fragments by 18 primers, the pairwise similarity indices between A. culbertsoni and other species (i.e., A. hatchetti, A. triangularis, A. polyphaga) were 0.300, 0.308, and 0.313, respectively. Similarity index between A. hatchetti and A. triangularis was 0.833. The mean similarity index among the 3 Korean isolates (YM-2, -3, -4) was 0.959 and 0.832 among them and 2 other species (A. hatchetti and A. triangularis). The mean similarity index among YM-5 and other Korean isolates (YM-2, -3, -4) was 0.237. However, the similarity index between YM-5 and A. culbertsoni was 0.857, which suggests that YM-5 is genetically more similar to A. culbertsoni than other Korean isolates. Phenogram reconstructed by UPGMA method revealed that there are two groups: one group consists of A. hatchetti, A. triangularis, and 3 Korean isolates (YM-2, -3, -4), and the other group consists of A. culbertsoni, A. polyphaga, HOV, and YM-5.

Figures


Fig. 1
Amplified DNA profiles of Acanthamoeba spp. using primers of 704 and 730. A. cul denotes A. culbertsoni, A. hat denotes A. hatchetti, A. tri denotes A. triangularis, A. pol denotes A. polyphaga, HOV denotes foreign isolate of HOV, Y2 denotes Korean isolate of YM-2, Y3 denotes Korean isolate of YM-3, Y4 denotes Korean isolate of YM-4, Y5 denotes Korean isolate of YM-5, L denotes DNA size marker of 100 base-pair ladder.


Fig. 2
Phenogram of Acanthamoeba spp. and isolates based on the dissimilarity estimates. Genetic distances were recalculated by UPGMA methods. Only the length of horizontal line, not vertical line, is meaningful for the genetic relationships among the operational taxonomic units.

Tables


Table 1
List of the Acanthamoeba spp. tested


Table 2
Arbitrary primers selected for full scale survey by RAPD-PCR analysis


Table 3
Pairwise dissimilarity estimates based on the RAPD markers generated by 18 arbitrary primers

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