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PHD : Parasites, Hosts and Diseases

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Original Article

Genetic analyses of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea

The Korean Journal of Parasitology 2001;39(2):161-170.
Published online: June 30, 2001

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Corresponding author (dichung@knu.ac.kr)
• Received: March 26, 2001   • Accepted: May 8, 2001

Copyright © 2001 by The Korean Society for Parasitology

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Genetic analyses of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea
Korean J Parasitol. 2001;39(2):161-170.   Published online June 30, 2001
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Genetic analyses of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea
Korean J Parasitol. 2001;39(2):161-170.   Published online June 30, 2001
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Genetic analyses of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea
Image Image Image Image Image
Fig. 1 Agarose gel electrophoretic restriction fragment patterns by EcoR I of mitochondrial DNA of Acanthamoeba 39 isolates. M, Hind III digested λ phage DNA as a DNA size standard.
Fig. 2 Agarose gel eletrophoretic restriction fragment patterns of mitochondrial DNA of seven different types of Acanthamoeba by eight kinds of restriction enzymes. M, Hind III digested λ phage DNA as a DNA size standard. Lane 1, KA/LS1; 2, KA/LS2; 3, KA/LS5; 4, KA/LS4; 5, KA/LS18; 6, KA/LS7; 7, KA/LS31.
Fig. 3 (A) Agarose gel electrophoretic pattern of PCR products of SSU rDNA of 7 types of Acanthamoeba isolates; M, Hind III digested λ phage DNA. (B)(C) Electrophoretic patterns of restriction fragments of PCR product from 7 types of Acanthamoeba isolates by Dde I and Hinf I; B, Agarose gel; C, Polyacrylamide gel; M, 50 bp ladder (BM, Germany); Lane 1, KA/LS1; 2, KA/LS2; 3, KA/LS5; 4, KA/LS4; 5, KA/LS18; 6. KA/LS7; 7, KA/LS31.
Fig. 4 UPGMA phenogram of 7 different types and 23 reference strains of Acanthamoeba based on the SSU rDNA PCR-RFLP analyses. ◎; 7 different types of Acanthamoeba isolates from contact lens storage cases.
Fig. 5 Comparative mitochondrial DNA RFLP patterns of Acanthamoeba isolated from contact lens storage cases with those of reference strains from keratitis. Lane 1, KA/LS1; 2, KA/E2; 3, KA/LS2; 4, Ma; 5, KA/LS5; 6, KA/E3; 7, KA/LS4; 8, KA/LS18; 9, KA/LS7; 10, KA/LS31; M, Hind III digested λ phage DNA.
Genetic analyses of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea
No. Strain ATCC No. Virulence Environmental source Geographic source Reference Former species designation
1 Castellani 30011 + yeast culture England Douglas (1930) A. castellanii
2 L3a 50240 + swimming pool France Pussard & Pons (1997) A. lugdunensis
3 Vil3 50241 nda) swimming pool France Pussard & Pons (1997) A. quina
4 Jones 30461 + keratitis U.S.A. Jones et al. (1975) A. polyphaga
5 SH621 50254 nd human feces France Pussard & Pons (1997) A. triangularis
6 Nagington 30873 + keratitis England Nagington et al. (1974) A. polyphaga
7 Singh 30973 soil England Singh (1952) A. rhysodes
8 1652 50253 soil Morocco Pussard & Pons (1977) A. mauritaniensis
9 AA2 50238 soil France Pussard & Pons (1977) A. divionensis
10 AA1 50251 soil France Pussard & Pons (1977) A. paradivionensis
11 Neff 30010 soil U.S.A. Neff (1957) A. castellanii
12 Ma 50370 + keratitis U.S.A. Ma et al. (1981) A. castellanii
13 P23 30871 fresh-water U.S.A. Page (1967) A. polyphaga
14 Chang 30898 + fresh water U.S.A. Byers et al.(1990) A. castellanii
15 BH-2 30730 + ocean sediment U.S.A. Sawyer et al.(1977) A. hatchetti
16 RB-F-1 50388 nd ocean sediment U.S.A. Sawyer et al. (1993) A. stevensoni
17 S-7 30731 + beach-bottom U.S.A. Sawyer (1971) A. griffini
18 Ray & Hayes 30137 nd soil U.S.A. Ray & Hayes (1954) A. astronyxis
19 OC-15C 30867 nd river U.S.A. Lewis & Sawyer (1979) A. tubiashi
20 A-1 30171 + tissue culture U.S.A. Singh & Das (1970) A. culbertsoni
21 OC-3A 30866 + GAEb) U.S.A. Moura et al. (1992) A. healyi
22 GE-3a 50252 swimming pool France Pussard & Pons (1977) A. pustulosa
23 Reich 30870 soil Israel Reich (1933) A. palestinensis
24 KA/E1 + keratitis Korea   —    —
25 KA/E2 + keratitis Korea   —    —
26 KA/E3 + keratitis Korea   —    —
27 KA/E4 + keratitis Korea   —    —
28 KA/E5 + keratitis Korea   —    —
29 KA/E6 + keratitis Korea   —    —
30 KA/E7 + keratitis Korea   —    —
31 KA/E8 + keratitis Korea   —    —
32 KA/E9 + keratitis Korea   —    —
33 KA/E10 + keratitis Korea   —    —
34 KA/E11 + keratitis Korea   —    —
35 KA/E12 + keratitis Korea   —    —
Isolates rDNA RFLP phenotype Mt DNA RFLP phenotype finally assigned species associated pathogenic strains
KA/LS1 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS2 KA/LS2 KA/LS2 A. castellanii Ma
KA/LS3 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS4 KA/LS4 KA/LS4 ?
KA/LS5 KA/LS5 KA/LS5 A. castellanii KA/E3, KA/E8
KA/LS6 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS7 KA/LS7 KA/LS7 ?
KA/LS8 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS9 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS10 KA/LS2 KA/LS2 A. castellanii Ma
KA/LS11 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/SL12 KA/LS2 KA/LS2 A. castellanii Ma
KA/LS13 KA/LS7 KA/LS7 ?
KA/LS14 KA/LS5 KA/LS5 A. castellanii KA/E3, KA/E8
KA/LS15 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS16 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS17 KA/LS2 KA/LS2 A. castellanii Ma
KA/LS18 KA/LS18 KA/LS18 A. castellanii JAC/E1a)
KA/LS19 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS20 KA/LS2 KA/LS2 A. castellanii Ma
KA/LS21 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS22 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS23 KA/LS2 KA/LS2 A. castellanii Ma
KA/LS24 KA/LS18 KA/LS18 A. castellanii JAC/E1
KA/LS25 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS26 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS27 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS28 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS29 KA/LS2 KA/LS2 A. castellanii Ma
KA/LS30 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS31 KA/LS31 KA/LS31 ?
KA/LS32 KA/LS2 KA/LS2 A. castellanii Ma
KA/LS33 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS34 KA/LS4 KA/LS4 ?
KA/LS35 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS36 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS37 KA/LS5 KA/LS5 A. castellanii KA/E3, KA/E8
KA/LS38 KA/LS1 KA/LS1 A. lugdunensis KA/E2, KA/E12
KA/LS39 KA/LS31 KA/LS31 ?
Type Seoul Southeast Southwest Total
KA/E2 20 (51.3%) 18 (64.3%) 8 (19.0%) 46 (43.8%)
KA/E3 3 (7.7%) 1 (3.6%) 21 (50.0%) 25 (23.8%)
A. castellanii Ma 8 (20.5%) 0 (0.0%) 9 (21.4%) 17 (16.2%)
Others 8 (20.5%) 9 (32.1%) 4 (9.5%) 17 (16.2%)
Total 39 28 42 105
Table 1. List of Acanthamoeba 35 reference strains

not determined

granulomatous amebic encephalitis

Table 2. Isolates of Acanthamoeba spp. from contact lens paraphernalias and restriction phenotypes of SSU rDNA and mitochondrial DNA

JAC/E1; pathogenic Acanthamoeba isolated from a patient with amebic keratitis in Japan (Yagita and Endo, 1990).

Table 3. Comparison of mtDNA RFLP type of Acanthamoeba isolates from contact lens paraphernalias of different areas of Korea