Warning: mkdir(): Permission denied in /home/virtual/lib/view_data.php on line 81
Warning: fopen(upload/ip_log/ip_log_2024-12.txt): failed to open stream: No such file or directory in /home/virtual/lib/view_data.php on line 83
Warning: fwrite() expects parameter 1 to be resource, boolean given in /home/virtual/lib/view_data.php on line 84 Differentiation of Korean isolates of Entamoeba histolytica from Entamoeba dispar
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
References
1.
Abd-Alla MD, Jackson TF, Gathiram V, el-Hawey AM, Ravdin JI. Differentiation of pathogenic Entamoeba histolytica infections from nonpathogenic infections by detection of galactose-inhibitable adherence protein antigen in sera and feces. J Clin Microbiol 1993;31(11):2845–2850.
2.
Blanc DS. Determination of taxonomic status of pathogenic and nonpathogenic Entamoeba histolytica zymodemes using isoenzyme analysis. J Protozool 1992;39(4):471–479.
3.
Chang JK, Im K, Soh CT. Axenization of Entamoeba histolytica, a Korean strain YS-27. Korean J Parasitol 1995;33(4):387–390.
4.
Clark CG, Diamond LS. Entamoeba histolytica: an explanation for the reported conversion of "nonpathogenic" amebae to the "pathogenic" form. Exp Parasitol 1993;77(4):456–460.
5.
Diamond LS, Clark CG. A redescription of Entamoeba histolytica Schaudinn, 1903 (Emended Walker, 1911) separating it from Entamoeba dispar Brumpt, 1925. J Eukaryot Microbiol 1993;40(3):340–344.
6.
Edman U, Meraz MA, Rausser S, Agabian N, Meza I. Characterization of an immuno-dominant variable surface antigen from pathogenic and nonpathogenic Entamoeba histolytica. J Exp Med 1990;172(3):879–888.
7.
Gonzalez-Ruiz A, Haque R, Rehman T, Aguirre A, Jaramillo C, Castanon G, Hall A, Guhl F, Ruiz-Palacios G, Warhurst DC, et al. A monoclonal antibody for distinction of invasive and noninvasive clinical isolates of Entamoeba histolytica. J Clin Microbiol 1992;30(11):2807–2813.
8.
Gonzalez-Ruiz A, Haque R, Rehman T, Aguirre A, Hall A, Guhl F, Warhurst DC, Miles MA. Diagnosis of amebic dysentery by detection of Entamoeba histolytica fecal antigen by an invasive strain-specific, monoclonal antibody-based enzyme-linked immunosorbent assay. J Clin Microbiol 1994;32(4):964–970.
9.
Haque R, et al. J Infect Dis 1993;167:2476–2479.
11.
Kappus KD, Lundgren RG Jr, Juranek DD, Roberts JM, Spencer HC. Intestinal parasitism in the United States: update on a continuing problem. Am J Trop Med Hyg 1994;50(6):705–713.
12.
Katzwinkel-Wladarsch S, Loscher T, Rinder H. Direct amplification and differentiation of pathogenic and nonpathogenic Entamoeba histolytica DNA from stool specimens. Am J Trop Med Hyg 1994;51(1):115–118.
14.
Lee SH. J Health Fell Found 1973;3:1–6.
15.
Lotter H, Mannweiler E, Schreiber M, Tannich E. Sensitive and specific serodiagnosis of invasive amebiasis by using a recombinant surface protein of pathogenic Entamoeba histolytica. J Clin Microbiol 1992;30(12):3163–3167.
16.
Mirelman D. Effect of culture conditions and bacterial associates on the zymodemes of Entamoeba histolytica. Parasitol Today 1987;3(2):37–40.
17.
Mirelman D, Bracha R, Chayen A, Aust-Kettis A, Diamond LS. Entamoeba histolytica: effect of growth conditions and bacterial associates on isoenzyme patterns and virulence. Exp Parasitol 1986;62(1):142–148.
18.
Mirelman D, Bracha R, Wexler A, Chayen A. Changes in isoenzyme patterns of a cloned culture of nonpathogenic Entamoeba histolytica during axenization. Infect Immun 1986;54(3):827–832.
19.
Tannich E, Burchard GD. Differentiation of pathogenic from nonpathogenic Entamoeba histolytica by restriction fragment analysis of a single gene amplified in vitro. J Clin Microbiol 1991;29(2):250–255.
20.
Tannich E, Horstmann RD, Knobloch J, Arnold HH. Genomic DNA differences between pathogenic and nonpathogenic Entamoeba histolytica. Proc Natl Acad Sci U S A 1989;86(13):5118–5122.
PDF Links
Full text via DOI
Download Citation 
