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Korean J Parasitol > Volume 30(4):1992 > Article

Original Article
Korean J Parasitol. 1992 Dec;30(4):289-298. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1992.30.4.289
Copyright © 1992 by The Korean Society for Parasitology
Immunological approach for classification of free-living amoeba in Korea
H J Shin,C H Kim and K I Im*
Department of Biology, College of Natural Science, Chungnam National University, Taejon 302-764, Korea.
Abstract

Acanthamoeba spp., free-living amoebae inhabited in moist soil, pond, freshwater, sewage, atmosphere and swimming pool, may be causative protozoa of the fatal primary amoebic meningoencephalitis in experimental animals and humans. In this study, Acanthamoeba spp., including Acanthamoeba sp. YM-4 (isolated strain from Korea) had been compared by the two-dimensional electrophoresis and hybridoma technique as well as the difference of morphological characteristics. Trophozoite of Acanthamoeba sp. YM-4 is usually uninucleate and show the hyaline filamentous projections (acanthopoda). No flagellate stage observed. Cysts have two walls, the outer wall is nearly circular, but inner wall is oval or some irregular. As results of SDS-PAGE for lysate of Acanthamoeba sp. YM-4, 16 major protein fractions are similar to those of A. culbertsoni, but different to A. royreba and A. polyphaga. Findings of two-dimensional electrophoretic patterns of Acanthamoeba sp. YM-4 are almost same to those of A. culbertsoni, The isotype of monoclonal antibodies produced from McAY 6, McAY 7, McAY 8, McAY 13 and McAY 16 clones were IgG1, and McAY 10 and McAY 11 clones were IgM. As results of the cross-reactivity among various amoebae using ELISA with monoclonal antibodies, McAY 7 monoclonal antibody (molecular weight 43 kDa by EITB) was only reacted with Acanthamoeba sp. YM-4, but McAY 6 and McAY 10 monoclonal antibodies were reacted to A. culbertsoni as well as Acanthamoeba sp. YM-4.

Figures


Fig. 1
Inverted microscopic findings of amoeba trophozoites(above) and cysts(below); Acanthamoeba sp. YM-4(a,b), A. culbertsoni(c,d), A. royreba(e,f) and A. polyphaga(g,h) (×1,000).


Fig. 2
SDS-PAGE patterns of Acanthamoeba sp. YM-4 (1) and reference species : A. culbertsoni(2), A. royreba(3) and A. polyphaga(4).


Fig. 3
Two-dimensional electrophoresis patterns of lysate from Acanthamoeba sp. YM-4 (1) and A. culbertsoni(B).


Fig. 4
Enzyme-linked immunoelectrotransfer blotting patterns of Acanthamoeba sp. YM-4 reacted with various monoclonal antibodies from cell lines McAY 7, McAY 8 and McAY 10; M.:protein standard, right: diagram of EITB.

Tables


Table 1
Immunoglobulin isotype of monoclonal antibodies against Acanthamoeba sp. YM-4


Table 2
Reactivity of anti-Acanthamoeba sp. YM-4 monoclonal antibodies with other free-living amoebae, measured by ELISA


Table 3
Classifical keys of genus Acanthamoeba based on the morphology of trophozoites and cysts

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