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Korean J Parasitol > Volume 34(4):1996 > Article

Original Article
Korean J Parasitol. 1996 Dec;34(4):247-254. English.
Published online Dec 20, 1996.  http://dx.doi.org/10.3347/kjp.1996.34.4.247
Copyright © 1996 by The Korean Society for Parasitology
Differentiation of Entamoeba histolytica and Entamoeba dispar in cyst-passers by immunoblot
Mejeong Lee and Sung-Tae Hong*
Department of Parasitology and Institute of Endemic Diseases, Seoul National University College of Medicine, Seoul, 110-799, Korea.
Received November 15, 1996; Accepted November 27, 1996.

Abstract

Differentiation of invasive strains of Entamoeba histolytica according to their pathogenicity has been a topic of long debate, but now the pathogenic species only is regarded as E. histolytica while the non-pathogenic species is E. dispar. The present study applied immunoblot to differentiate infections of the two species among microscopically-detected cyst-passers in Korea. The crude extract of E. histolytica separated in 5-20% gradient gels, revealed many fractions of 94, 81, 71, 50, 44, 38.5, 37.5, 29, 19, and 18 kDa when the cysteine proteinase inhibitor, E64, was supplemented. The serum IgG antibody of proven E. histolytica cases reacted with the antigenic fractions of 117, 110, 99, 68, 66, 60, 54, 52, 46, and 45 kDa. Sera of PCR confirmed 3 cases of E. dispar reacted only to the 117 kDa fraction of the E. histolytica crude extract which was regarded as non-specific. To the antigen of monoxenic E. dispar, sera of E. dispar and E. histolytica cases showed the same immunoblot reactions. The serum IgA antibody reacted with several antigenic fractions of both E. histolytica and E. dispar, but IgM and IgE antibodies showed no reaction to either antigen. Sera of 24 symptomless amebic cyst-passers were screened with the E. histolytica antigen; two were found to be infected by E. histolytica and 22 were by E. dispar. The present findings suggest that in Korea most asymptomatic cyst passers of E. histolytica are carriers of E. dispar. Immunoblot using E. histolytica antigen is a good technique for the differentiation of E. histolytica and E. dispar infections.

Figures


Fig. 1
The effects of proteinase inhibitors on crude extracts of E. histolytica YS-27 trophozoites by SDS-PAGE in a 5-20% gradient gel. Lane 1. serine proteinase inhibitor (PMSF); lane 2, aspartic proteinase inhibitor (pepstatin A); lane 3, cysteine proteinase inhibitor (E64); lane 4, cysteine proteinase inhibitor (iodoacetic acid); lane 5, without proteinase inhibitors.


Fig. 2
Comparison of crude extracts of E. histolytica and E. dispar separated in 5-20% gradient SDS-PAGE. Lane 1, E. histolytica (YS-29); lane 2, E. dispar (S16); lane 3, Escherichia coli.


Fig. 3
Immunoblot patterns of serum IgG antibodies to the E64 supplemented crude extracts of E. histolytica. Lane 1, amido black 10B-stained protein fractions; lanes 2-4, sera of E. histolytica patients; lanes 5-8, sera of E. dispar infected humans; lane 9, serum of E. coli infection; lane 10, serum of B. hominis infection; lane 11, serum of normal control.


Fig. 4
Immunoblot patterns of serum IgA antibodies to the E64 supplemented crude extracts of E. histolytica. Lane 1, amido black 10B-stained protein fractions; lanes 2-4, sera of E. histolytica patients; lanes 5-8, sera of E. dispar infected humans; lane 9, serum of E. coli infection; lane 10, serum of B. hominis infection; lane 11, serum of a normal control.


Fig. 5
Immunoblot patterns of serum IgG antibodies to the E64 supplemented crude extracts of E. dispar. Lanes 1-3, sera of E. histolytica infection; lanes 4-6, sera of E. dispar infection; lane 7, serum of a control.


Fig. 6
Immunoblot patterns of serum IgA antibodies to the E64 supplemented crude extracts of E. dispar. Lanes 1-3, sera of E. histolytica infection; lanes 4-6, sera of E. dispar infection; lane 7, control serum.


Fig. 7
Screening of 24 symptomless cyst-passers with antigen of E. histolytica. The sera probed to the strips 11 and 12 were regarded as infection of E. histolytica, and all of ohers were of E. dispar.

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