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. 1991 Apr;29(4):696–701. doi: 10.1128/jcm.29.4.696-701.1991

Isolation of a strain-specific Entamoeba histolytica cDNA clone.

D J Burch 1, E Li 1, S Reed 1, T F Jackson 1, S L Stanley Jr 1
PMCID: PMC269855  PMID: 1890170

Abstract

Entamoeba histolytica is an intestinal parasite causing significant morbidity and mortality worldwide. More tools are needed to understand the epidemiology and molecular pathogenesis of amebiasis. A cDNA library was constructed by using poly(A)+ RNA isolated from an axenic strain of E. histolytica, HM1:IMSS, which expresses a pathogenic isoenzyme pattern (zymodeme). Differential screening of the library yielded a strain-specific 3' polyadenylated cDNA clone, C2, possessing nine 26-nucleotide tandem repeats. RNA and DNA transfer blot analysis of four axenic strains of E. histolytica possessing the same pathogenic zymodeme revealed that the gene is present and expressed in pathogenic E. histolytica HM1:IMSS and 200:NIH but is not present in pathogenic strains HK-9 and Rahman. In addition, Southern blot analysis using the C2 clone showed heterogeneity of genomic organization between HM1:IMSS and 200:NIH. DNA dot blot hybridization analysis demonstrated that cDNA clone C2 was also able to distinguish axenically cultured E. histolytica strains possessing pathogenic zymodemes from those possessing nonpathogenic zymodemes and could detect as few as 100 amebic trophozoites. We conclude that C2 is a strain-specific E. histolytica cDNA clone that, in conjunction with other E. histolytica-specific probes, could serve as a useful epidemiologic tool.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bracha R., Diamond L. S., Ackers J. P., Burchard G. D., Mirelman D. Differentiation of clinical isolates of Entamoeba histolytica by using specific DNA probes. J Clin Microbiol. 1990 Apr;28(4):680–684. doi: 10.1128/jcm.28.4.680-684.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Diamond L. S., Harlow D. R., Cunnick C. C. A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans R Soc Trop Med Hyg. 1978;72(4):431–432. doi: 10.1016/0035-9203(78)90144-x. [DOI] [PubMed] [Google Scholar]
  3. Edman U., Meza I., Agabian N. Genomic and cDNA actin sequences from a virulent strain of Entamoeba histolytica. Proc Natl Acad Sci U S A. 1987 May;84(9):3024–3028. doi: 10.1073/pnas.84.9.3024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  5. Garfinkel L. I., Giladi M., Huber M., Gitler C., Mirelman D., Revel M., Rozenblatt S. DNA probes specific for Entamoeba histolytica possessing pathogenic and nonpathogenic zymodemes. Infect Immun. 1989 Mar;57(3):926–931. doi: 10.1128/iai.57.3.926-931.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Guerrant R. L. Amebiasis: introduction, current status, and research questions. Rev Infect Dis. 1986 Mar-Apr;8(2):218–227. doi: 10.1093/clinids/8.2.218. [DOI] [PubMed] [Google Scholar]
  7. Li E., Becker A., Stanley S. L., Jr Chinese hamster ovary cells deficient in N-acetylglucosaminyltransferase I activity are resistant to Entamoeba histolytica-mediated cytotoxicity. Infect Immun. 1989 Jan;57(1):8–12. doi: 10.1128/iai.57.1.8-12.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Mattern C. F., Keister D. B., Caspar P. A. Experimental amebiasis. III. A rapid in vitro assay for virulence of Entamoeba histolytica. Am J Trop Med Hyg. 1978 Sep;27(5):882–887. [PubMed] [Google Scholar]
  9. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mirelman D., Bracha R., Chayen A., Aust-Kettis A., Diamond L. S. Entamoeba histolytica: effect of growth conditions and bacterial associates on isoenzyme patterns and virulence. Exp Parasitol. 1986 Aug;62(1):142–148. doi: 10.1016/0014-4894(86)90017-2. [DOI] [PubMed] [Google Scholar]
  11. Robinson G. L. The laboratory diagnosis of human parasitic amoebae. Trans R Soc Trop Med Hyg. 1968;62(2):285–294. doi: 10.1016/0035-9203(68)90170-3. [DOI] [PubMed] [Google Scholar]
  12. Samuelson J., Acuna-Soto R., Reed S., Biagi F., Wirth D. DNA hybridization probe for clinical diagnosis of Entamoeba histolytica. J Clin Microbiol. 1989 Apr;27(4):671–676. doi: 10.1128/jcm.27.4.671-676.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Sargeaunt P. G. The reliability of Entamoeba histolytica zymodemes in clinical diagnosis. Parasitol Today. 1987 Feb;3(2):40–37. doi: 10.1016/0169-4758(87)90211-0. [DOI] [PubMed] [Google Scholar]
  14. Sargeaunt P. G., Williams J. E., Grene J. D. The differentiation of invasive and non-invasive Entamoeba histolytica by isoenzyme electrophoresis. Trans R Soc Trop Med Hyg. 1978;72(5):519–521. doi: 10.1016/0035-9203(78)90174-8. [DOI] [PubMed] [Google Scholar]
  15. Stanley S. L., Jr, Becker A., Kunz-Jenkins C., Foster L., Li E. Cloning and expression of a membrane antigen of Entamoeba histolytica possessing multiple tandem repeats. Proc Natl Acad Sci U S A. 1990 Jul;87(13):4976–4980. doi: 10.1073/pnas.87.13.4976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Strachan W. D., Chiodini P. L., Spice W. M., Moody A. H., Ackers J. P. Immunological differentiation of pathogenic and non-pathogenic isolates of Entamoeba histolytica. Lancet. 1988 Mar 12;1(8585):561–563. doi: 10.1016/s0140-6736(88)91355-4. [DOI] [PubMed] [Google Scholar]
  17. Tannich E., Horstmann R. D., Knobloch J., Arnold H. H. Genomic DNA differences between pathogenic and nonpathogenic Entamoeba histolytica. Proc Natl Acad Sci U S A. 1989 Jul;86(13):5118–5122. doi: 10.1073/pnas.86.13.5118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]

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