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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
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

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.

Type
Research Article
Copyright
2004 Cambridge University Press

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