Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 May;35(5):1108–1111. doi: 10.1128/jcm.35.5.1108-1111.1997

An improved colorimetric PCR-based method for detection and differentiation of Entamoeba histolytica and Entamoeba dispar in feces.

D Britten 1, S M Wilson 1, R McNerney 1, A H Moody 1, P L Chiodini 1, J P Ackers 1
PMCID: PMC232712  PMID: 9114390

Abstract

The epidemiological implications of the recent separation of "Entamoeba histolytica" into two separate species, pathogenic E. histolytica sensu stricto and commensal E. dispar, will not become apparent without methods of distinguishing between them which are applicable to large numbers of specimens. We have modified a PCR-based method to produce such a technique which may be completed in 1 day while still identifying 10(-1) E. histolytica and 1 to 10 E. dispar trophozoites per g of feces when present separately and 10 E. histolytica and 100 E. dispar trophozoites per g in the presence of 10(6) trophozoites per g of the other species. Applied to fecal specimens from 18 patients from which E. histolytica or E. dispar had been grown and identified to the species level by hexokinase isoenzyme analysis, the method in every case yielded the correct result. Positive and negative results are easily distinguished by eye, and we are now applying this technique to a large-scale epidemiological study of amebiasis in the eastern Mediterranean region.

Full Text

The Full Text of this article is available as a PDF (191.9 KB).

Selected References

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

  1. Abd-Alla M. D., Jackson T. F., Gathiram V., el-Hawey A. M., Ravdin J. I. 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 Nov;31(11):2845–2850. doi: 10.1128/jcm.31.11.2845-2850.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Acuna-Soto R., Samuelson J., De Girolami P., Zarate L., Millan-Velasco F., Schoolnick G., Wirth D. Application of the polymerase chain reaction to the epidemiology of pathogenic and nonpathogenic Entamoeba histolytica. Am J Trop Med Hyg. 1993 Jan;48(1):58–70. doi: 10.4269/ajtmh.1993.48.58. [DOI] [PubMed] [Google Scholar]
  3. Aguirre A., Warhurst D. C., Guhl F., Frame I. A. Polymerase chain reaction-solution hybridization enzyme-linked immunoassay (PCR-SHELA) for the differential diagnosis of pathogenic and non-pathogenic Entamoeba histolytica. Trans R Soc Trop Med Hyg. 1995 Mar-Apr;89(2):187–188. doi: 10.1016/0035-9203(95)90488-3. [DOI] [PubMed] [Google Scholar]
  4. Allen A. V., Ridley D. S. Further observations on the formol-ether concentration technique for faecal parasites. J Clin Pathol. 1970 Sep;23(6):545–546. doi: 10.1136/jcp.23.6.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boom R., Sol C. J., Salimans M. M., Jansen C. L., Wertheim-van Dillen P. M., van der Noordaa J. Rapid and simple method for purification of nucleic acids. J Clin Microbiol. 1990 Mar;28(3):495–503. doi: 10.1128/jcm.28.3.495-503.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clark C. G., Diamond L. S. Entamoeba histolytica: a method for isolate identification. Exp Parasitol. 1993 Dec;77(4):450–455. doi: 10.1006/expr.1993.1105. [DOI] [PubMed] [Google Scholar]
  7. Diamond L. S., Clark C. G. A redescription of Entamoeba histolytica Schaudinn, 1903 (Emended Walker, 1911) separating it from Entamoeba dispar Brumpt, 1925. J Eukaryot Microbiol. 1993 May-Jun;40(3):340–344. doi: 10.1111/j.1550-7408.1993.tb04926.x. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Haque R., Neville L. M., Wood S., Petri W. A., Jr Short report: detection of Entamoeba histolytica and E. dispar directly in stool. Am J Trop Med Hyg. 1994 May;50(5):595–596. doi: 10.4269/ajtmh.1994.50.595. [DOI] [PubMed] [Google Scholar]
  11. 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 Jul;51(1):115–118. doi: 10.4269/ajtmh.1994.51.115. [DOI] [PubMed] [Google Scholar]
  12. McNerney R., Frame I. A., Vexenat J. A., Fonseca de Castro J. A., Howard M. K., Dillon R., Wilson S., Miles M. A. Visceral leishmaniasis in Teresina, n. e. Brazil: towards a DNA probe kit and its adaptation to processing blood-contaminated samples. Arch Inst Pasteur Tunis. 1993 Jul-Oct;70(3-4):405–418. [PubMed] [Google Scholar]
  13. Rivera W. L., Tachibana H., Silva-Tahat M. R., Uemura H., Kanbara H. Differentiation of Entamoeba histolytica and E. dispar DNA from cysts present in stool specimens by polymerase chain reaction: its field application in the Philippines. Parasitol Res. 1996;82(7):585–589. doi: 10.1007/s004360050169. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Romero J. L., Descoteaux S., Reed S., Orozco E., Santos J., Samuelson J. Use of polymerase chain reaction and nonradioactive DNA probes to diagnose Entamoeba histolytica in clinical samples. Arch Med Res. 1992;23(2):277–279. [PubMed] [Google Scholar]
  16. Sehgal R., Abd-Alla M., Moody A. H., Chiodini P. L., Ackers J. P. Comparison of two media for the isolation and short-term culture of Entamoeba histolytica and E. dispar. Trans R Soc Trop Med Hyg. 1995 Jul-Aug;89(4):394–394. doi: 10.1016/0035-9203(95)90023-3. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Tachibana H., Kobayashi S., Okuzawa E., Masuda G. Detection of pathogenic Entamoeba histolytica DNA in liver abscess fluid by polymerase chain reaction. Int J Parasitol. 1992 Dec;22(8):1193–1196. doi: 10.1016/0020-7519(92)90042-j. [DOI] [PubMed] [Google Scholar]
  19. Walsh J. A. Problems in recognition and diagnosis of amebiasis: estimation of the global magnitude of morbidity and mortality. Rev Infect Dis. 1986 Mar-Apr;8(2):228–238. doi: 10.1093/clinids/8.2.228. [DOI] [PubMed] [Google Scholar]
  20. Wilson S. M., McNerney R., Nye P. M., Godfrey-Faussett P. D., Stoker N. G., Voller A. Progress toward a simplified polymerase chain reaction and its application to diagnosis of tuberculosis. J Clin Microbiol. 1993 Apr;31(4):776–782. doi: 10.1128/jcm.31.4.776-782.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES