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A single step duplex PCR to distinguish Entamoeba histolytica from Entamoeba dispar

Published online by Cambridge University Press:  13 May 2004

M. A. R. FREITAS ,
E. N. VIANNA ,
A. S. MARTINS ,
E. F. SILVA ,
J. L. PESQUERO  and
M. A. GOMES
M. A. R. FREITAS
Affiliation:
Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, CEP: 31270-901 Belo Horizonte, MG, Brasil
E. N. VIANNA
Affiliation:
Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, CEP: 31270-901 Belo Horizonte, MG, Brasil
A. S. MARTINS
Affiliation:
Departamento de Fisiologia e Biofisica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, CEP: 31270-901 Belo Horizonte, MG, Brasil
E. F. SILVA
Affiliation:
Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, CEP: 31270-901 Belo Horizonte, MG, Brasil
J. L. PESQUERO
Affiliation:
Departamento de Fisiologia e Biofisica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, CEP: 31270-901 Belo Horizonte, MG, Brasil Universidade de Mogi das Cruzes, São Paulo, SP, Brasil
M. A. GOMES
Affiliation:
Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, CEP: 31270-901 Belo Horizonte, MG, Brasil
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Abstract

In this study, a single-step duplex polymerase chain reaction procedure was developed for rapid, specific and sensitive identification of Entamoeba histolytica and for its diagnostic differentiation from E. dispar. Specific oligonucleotide primers were combined for the amplification of a cysteine proteinase 5 gene target sequence of 242 bp, present only in E. histolytica. Additionally, another oligonucleotide primer pair for both the E. histolytica and E. dispar actin gene target of 300 bp was designed to amplify only from amoebae DNA. The PCR developed was specific and efficiently identified and differentiated these parasites from each other in either cultured parasites or from stool material.

Keywords

Entamoeba histolytica–Entamoeba dispar diagnosisPCRactincysteine proteinase 5
Type
Research Article
Information
Parasitology , Volume 128 , Issue 6 , June 2004 , pp. 625 - 628
Copyright
2004 Cambridge University Press

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References

REFERENCES

ALI, I. K., HOSSAIN, M. B., ROY, S., AYEH-KUMI, P. F., PETRI, W. A. Jr., HAQUE, R. & CLARK, C. G. ( 2003). Entamoeba moshkovskii infections in children, Bangladesh. Emerging Infectious Diseases 9, 580–584.CrossRefGoogle Scholar
BLESSMANN, J., BUSS, H., TON NU, P. A., DINH, B. T., VIET NGO, Q. T., VAN, A. L., ABD ALLA, M. D., JACKSON, T. F. H. G., RAVDIN, J. I. & TANNICH, E. ( 2002). Real-time PCR for detection and differentiation of Entamoeba histolytica and Entamoeba dispar in fecal samples. Journal of Clinical Microbiology 40, 4413–4417.CrossRefGoogle Scholar
BRITTEN, D., WILSON, S. M., McNERNEY, R., MOODY, A. H., CHIODINI, P. L. & ACKERS, J. P. ( 1997). An improved colorimetric PCR-based method for detection and differentiation of Entamoeba histolytica and Entamoeba dispar in feces. Journal of Clinical Microbiology 35, 1108–1111.Google Scholar
BRUCHHAUS, I., JACOBS, T., LEIPPE, M. & TANNICH, E. ( 1996). Entamoeba histolytica and E. dispar: differences in numbers and expression of cysteine proteinase genes. Molecular Microbiology 22, 255–263.Google Scholar
BRUCHHAUS, I., ROEDER, T., LOTTER, H., SCHWERDTFEGER, M. & TANNICH, E. ( 2002). Differential gene expression in Entamoeba histolytica isolated from amoebic liver abscess. Molecular Microbiology 44, 1063–1072.CrossRefGoogle Scholar
DIAMOND, L. S., HARLOW, D. R. & CUNNICK, C. C. ( 1978). A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Transactions of the Royal Society of Tropical Medicine and Hygiene 86, 170–176.CrossRefGoogle Scholar
EDMAN, U., MEZA, I. & AGABIAN, N. ( 1987). Genomic and cDNA actin sequences from a virulent strain of Entamoeba histolytica. Proceedings of the National Academy of Sciences, USA 84, 3024–3028.CrossRefGoogle Scholar
EDMAN, U., MERAZ, M. A., RAUSSER, S., AGABIAN, N. & MEZA, I. ( 1990). Characterization of an immuno-dominant variable surface antigen from pathogenic and nonpathogenic Entamoeba histolytica. Journal of Experimental Parasitology 172, 879–888.CrossRefGoogle Scholar
GOMES, M. A., PESQUERO, J. B., FURST, C., VALLE, P. R., PESQUERO, J. L. & SILVA, E. F. ( 1999). An improved method to distinguish Entamoeba histolytica and Entamoeba dispar. Parasitology 119, 359–362.CrossRefGoogle Scholar
HAQUE, R. L., NEVILLE, L. M., HAHN, P. & PETRI, W. A. Jr. ( 1995). Rapid diagnosis of Entamoeba infection by using Entamoeba and Entamoeba histolytica stool antigen detection kits. Journal of Clinical Microbiology 33, 2558–2561.Google Scholar
HELLBERG, A., NICKEL, R., LOTTER, H., TANNICH, E. & BRUCHHAUS, I. ( 2001). Overexpression of cysteine proteinase 2 in Entamoeba histolytica or Entamoeba dispar increases amoeba-induced monolayer destruction in vitro but does not augment amoebic liver abscess formation in gerbils. Cellular Microbiology 3, 13–20.CrossRefGoogle Scholar
HUBER, M., GARFINKEL, L., GITLER, C., MIRELMAN, D., REVEL, M. & ROZEMBLATT, S. ( 1987). Entamoeba histolytica cloning and characterization of actin cDNA. Molecular and Biochemical Parasitology 24, 227–235.CrossRefGoogle Scholar
JACOBS, T., BRUCHHAUS, I., DANDEKAS, T., TANNICH, E. & LEIPE, M. ( 1998). Isolation and molecular characterization of a surface-bound proteinase of Entamoeba histolytica. Molecular Microbiology 27, 269–276.CrossRefGoogle Scholar
NÚÑEZ, Y. O., FERNANDEZ, M. A., TORREZ-NUNEZ, D., SILVA, J. A., MONTANO, I., MAESTRE, J. L. & FONTE, L. ( 2001). Multiplex polymerase chain reaction amplification and differentiation of Entamoeba histolytica and Entamoeba dispar DNA from stool samples. American Journal of Tropical Medicine and Hygiene 64, 293–297.CrossRefGoogle Scholar
SILVA, E. F. & MAYRINK, W. ( 1974). Estudos sobre a Entamoeba moshkovskii. Novos focos em diversos tipos de coleções hídricas no Brasil e Uruguai. Revista do Instituto de Medicina Tropical de São Paulo 16, 203–221.Google Scholar
RIDLEY, D. S. & HAWGOOD, B. C. ( 1956). The value of formol–ether concentration of faecal cysts and ova. Journal of Clinical Pathology 9, 74–76.CrossRefGoogle Scholar
REED, S. L., EMBER, J. A., HERDMAN, D. S., DISAPIO, R. G., HUGLI, T. E. & GIGLI, I. ( 1995). The extracellular neutral cysteine proteinase of Entamoeba histolytica degrades anaphylatoxins C3a and C5a. Journal of Immunology 155, 266–274.Google Scholar
SARGEAUNT, P. G., WILLIAMS, J. E. & GRENE, J. D. ( 1978). The differentiation of invasive and non-invasive Entamoeba histolytica by isoenzyme electrophoresis. Transactions of the Royal Society of Tropical Medicine and Hygiene 72, 519–521.CrossRefGoogle Scholar
VERWEIJ, J. J., BLOTKAMP, J., BRIENEN, E. A., AGUIRRE, A. & POLDERMAN, A. M. ( 2000). Differentiation of Entamoeba histolytica and Entamoeba dispar cysts using polymerase chain reaction on DNA isolated from faeces with spin columns. European Journal of Clinical Microbiology and Infectious Diseases 19, 358–361.CrossRefGoogle Scholar
WALSH, J. A. ( 1986). Problems in recognition and diagnosis of amoebiasis. Estimation of the global magnitude of morbidity and mortality. Reviews of Infectious Diseases 66, 228–238.Google Scholar
WHO/PAN AMERICA HEALTH ORGANIZATION ( 1997). Expert consulation on amoebiasis: Amoebiasis. WHO Weekly Epidemiological Record 72, 97–100.
WILLHOEFT, U., HAMANN, L. & TANNICH, E. ( 1999). A DNA sequence corresponding to the gene encoding cysteine proteinase 5 in Entamoeba histolytica is present and positionally conserved but highly degenerated in Entamoeba dispar. Infection and Immunity 67, 5925–5929.Google Scholar