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Differentiation of Korean isolates of Entamoeba histolytica from Entamoeba dispar
Seong-Choon Choe,1Mejeong Lee,1Sang-Kum Lee,2Kyung-il Im,3Egbert Tannich,4Soon-Hyung Lee,1 and Sung-Tae Hong*1
1Department of Parasitology and Institute of Endemic Diseases, Seoul National University College of Medicine, Seoul 110-799, Korea.
2Department of Clinical Pathology, Seoul Paik Hospital, Seoul 100-031, Korea.
3Department of Parasitology, Yonsei University College of Medicine, Seoul 120-752, Korea.
4Bernhard Nocht Institute for Tropical Medicine, 20359 Hamburg, Germany.
Received January 15, 1996; Accepted February 01, 1996.
Abstract
Cysts of Entamoeba histolytica are still found from humans in Korea, but not all of the cysts are known as pathogenic. The non-pathogenic strain is regarded as a different species, E. dispar. In this study, Korean isolates of conventional E. histolytica were subjected for the differentiation by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis. Human stools were screened by routine microscopic examination, and cyst or trophozoite positive stools were inoculated into Robinson media. The cultivated trophozoite positive stools were inoculated into Robinson media. The cultivated trophozoites were prepared for DNA extraction, and the DNAs were used for PCR with common primers of P1 gene. The PCR products were digested with 3 restriction enzymes and RFLP was observed. Also anti- sense primers containing the cleavage site of each restriction enzyme were designed for differentiation only by PCR. The PCR products of Korean isolates S9, S12, YS-6, and YS-27 were spliced by Taq I and Xmn I but not by Acc I, and the isolates S1, S3, S11, S15, S16, S17, S20, YS-17, and YS-44 were spliced by Acc I but not by Taq I and Xmn I. These RFLP pattern correlated well with PCR products by the species specific primers. The findings confirm that the Korean isolates S9, S12, YS-6, and YS-27 are E. histolytica and others are E. dispar. In Korea, most of the asymptomatic cyst carriers are infected by E. dispar, not by E. histolytica.
Figures
Fig. 1 PCR products of HM-1 & S12 (E. histolytica) and S15 (E. dispar) with the primer pairs of P1-S17 and 3 antisense primers. 1-3, HM-1 amoeba template DNA with primers of comnon P1-AS20, pathogenic P1-AS16, and non-pothogenic NP1-AS16; 4-6, S12 amoeba template DNA with primers of common P1-AS20, pathogenic P1-AS16, and non-pathogenic NP1-AS16; 7-9, S15 amoeba template DNA with primers of common P1-AS20, pathogenic P1-AS16, and non-pathogenic P1-NAS16.
Fig. 2 Restriction patterns of 482 bp PCR products of S12 (E. histolytica) and S15 (E. dispar) template DNAs with common primers of P1-S17 and P1-AS20. S12 product was digested into two fragments; 321 and 161 bp by T, and 291 and 191 bp by X. S15 product was digested into two fragments, 292 and 190 bp by A. Restriction enzymes: T, Taq I; X, Xmn I; A, Acc I.
Tables
Table 1 Differentiation of E. histolytica and E. dispar among 13 Korean Entamoeba isolates
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