The pathogenicity of 47 strains ofEntamoeba histolytica isolated in Pernambuco, Brazil, was examined using the polymerase chain reaction (PCR) followed by restriction-endonuclease digestion. Electrophoretic patterns of PCR products digested withHinfI revealed that all strains were nonpathogenic. The results were entirely in accord with phenotypic properties such as isoenzyme patterns and the failure to bind a pathogenic-isolatespecific monoclonal antibody. When the sensitivity of PCR was examined, amplified products could be detected from template DNA equivalent to five trophozoites. These observations indicate that PCR amplification of genomic DNA and subsequent restriction-enzyme digestion is a useful strategy for obtaining a sensitive and accurate diagnosis. The present study also demonstrates that nonpathogenic strains ofE. histolytica predominate in northeastern Brazil.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Andrews BJ, Mentzoni L, Bjorvatn B (1990) Zymodeme conversion of isolates ofEntamoeba histolytica. Trans R Soc Trop Med Hyg 84:63–65
Blanc D, Sargeaunt PG (1991)Entamoeba histolytica zymodemes: exhibition of γ and δ bands only of glucose phosphate isomerase and phosphoglucomutase may be influenced by starch content in the medium. Exp Parasitol 72:87–90
Bracha R, Diamond LS, Ackers JP, Burchard GD, Mirelman D (1990) Differentiation of clinical isolates ofEntamoeba histolytica by using specific DNA probes. J Clin Microbiol 28:680–684
Burch DJ, Li E, Reed S, Jackson TFHG, Stanley SL Jr (1991) Isolation of strain-specificEntamoeba histolytica cDNA clone. J Clin Microbiol 29:696–701
Edman U, Meraz MA, Rausser S, Agabian N, Meza I (1990) Characterization of an immunodominant variable surface antigen from pathogenic and nonpathogenicEntamoeba histolytica. J Exp Med 172:879–888
Garfinkel LI, Giladi M, Huber M, Gitler C, Mirelman D, Revel M, Rozenblatt S (1989) DNA probes specific forEntamoeba histolytica possessing pathogenic and nonpathogenic zymodemes. Infect Immun 57:926–931
Gonçalves JF, Tanabe M, Medeiros FPM, Gonçalves FJ, Aca IS, Motta SRN, Tateno S, Takeuchi T (1990) Parasitological and serological studies on amoebiasis and other intestinal parasitic infections in the rural sector around Recife, northeast Brazil. Rev Inst Med Trop Sao Paulo 32:428–435
Mirelman D (1987) Effect of culture conditions and bacterial associates on the zymodemes ofEntamoeba histolytica. Parasitol Today 3:37–40
Nozaki T, Aca IS, Okuzawa E, Magalhães M, Tateno S, Takeuchi T (1990) Zymodemes ofEntamoeba histolytica isolated in the Amazon and the north-east of Brazil Trans R Soc Trop Med Hyg 84:387–388
Okazaki M, Miranda P, Neto J, Diegues V, Alves J, Cauas M, Tanabe M, Kobayashi S, Kaneko N, Nagakura K, Kobayashi M, Motta S, Tateno S, Takeuchi T (1988) Parasitological and serological studies on amoebiasis and other intestinal parasitic infection in Recife and its suburban area, northeast Brazil. Rev Inst Med Trop Sao Paulo 30:313–321
Robinson GL (1968) The laboratory diagnosis of human parasitic amoebae. Trans R Soc Trop Med Hyg 62:285–294
Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988) Primer-directed enzyme amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
Sargeaunt PG (1988) Zymodemes ofEntamoeba histolytica. In: Ravdin JI (ed) Amebiasis: human infection byEntamoeba histolytica. John Wiley & Sons, New York, pp 370–387
Sargeaunt PG, Baveja UK, Nanda R, Anand BS (1984) Influence of geographical factors in the distribution of pathogenic zymodemes ofEntamoeba histolytica: identification of zymodeme XIV in India. Trans R Soc Trop Med Hyg 78:96–101
Tachibana H, Kobayashi S, Kato Y, Nagakura K, Kaneda Y, Takeuchi T (1990) Identification of a pathogenic isolate-specific 30,000-Mr antigen ofEntamoeba histolytica by using a monoclonal antibody. Infect Immun 58:955–960
Tachibana H, Ihara S, Kobayashi S, Kaneda Y, Takeuchi T, Watanabe Y (1991a) Differences in genomic DNA sequences between pathogenic and nonpathogenic isolates ofEntamoeba histolytica identified by polymerase chain reaction. J Clin Microbiol 29:2234–2239
Tachibana H, Kobayashi S, Nagakura K, Kaneda Y, Takeuchi T (1991b) Reactivity of monoclonal antibodies to species-specific antigens ofEntamoeba histolytica. J Protozool 38:329–334
Takeuchi T, Kobayashi S (1983) Serological diagnosis of amoebiasis. Immunoadvance 12:27–30
Tanabe M, Okazaki M, Okazaki M, Kobayashi S, Kaneko N, Sekiguchi T, Tateno S, Motta SRN, Takeuchi T (1990) Serological studies on schistosomiasis mansoni in the northeast of Brazil (I). Rev Inst Med Trop Sao Paulo 32:121–131
Tannich E, Burchard GD (1991) Differentiation of pathogenic from nonpathogenicEntamoeba histolytica by restriction fragment analysis of a single gene amplified in vitro. J Clin Microbiol 29:250–255
Tannich E, Horstmann RD, Knobloch J, Arnold HH (1989) Genomic DNA differences between pathogenic and nonpathogenicEntamoeba histolytica. Proc Natl Acad Sci USA 86:5118–5122
Tannich E, Scholze H, Nickel R, Horstmann RD (1991) Homologous cysteine proteinases of pathogenic and nonpathogenicEntamoeba histolytica. J Biol Chem 266:4798–4803
Walsh JA (1988) Prevalence ofEntamoeba histolytica infection. In: Ravdin JI (ed) Amebiasis: human infection byEntamoeba histolytica. John Wiley & Sons, New York, pp 93–105
This study was supported by the Japan International Cooperation Agency (JICA), by a Tokai University School of Medicine Research Aid award, and by a grant from the Ohyama Health Foundation
About this article
Cite this article
Tachibana, H., Kobayashi, S., Paz, K.C. et al. Analysis of pathogenicity by restriction-endonuclease digestion of amplified genomic DNA ofEntamoeba histolytica isolated in Pernambuco, Brazil. Parasitol Res 78, 433–436 (1992). https://doi.org/10.1007/BF00931701
- Polymerase Chain Reaction
- Monoclonal Antibody
- Polymerase Chain Reaction Product
- Polymerase Chain Reaction Amplification
- Accurate Diagnosis