A single step duplex PCR to distinguish Entamoeba histolytica from Entamoeba dispar.
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.
Freitas
M A R
MA
Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, CEP: 31270-901 Belo Horizonte, MG, Brasil.
Vianna
E N
EN
Martins
A S
AS
Silva
E F
EF
Pesquero
J L
JL
Gomes
M A
MA
D000199:Actins / Q000737:chemistry / Q000235:genetics; D000818:Animals; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000235:genetics; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D004404:Dysentery, Amebic / Q000469:parasitology*; D004748:Entamoeba histolytica / Q000145:classification / Q000201:enzymology / Q000235:genetics*; D005243:Feces / Q000469:parasitology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity
D000199:Actins; D016054:DNA, Protozoan; D003546:Cysteine Endopeptidases