title: Multiplex polymerase chain reaction amplification and differentiation of Entamoeba histolytica and Entamoeba dispar DNA from stool samples.
abstract: Due to the clinical importance of differentiating the two species of the Entamoeba histolytica/Entamoeba dispar complex, we developed a multiplex polymerase chain reaction (PCR) method that overcomes time-consuming and laborious procedures. We report here a DNA extraction protocol using non-fixed stool samples that avoid long lysis-incubation periods through the combined use of zirconium beads and a lysis-supporting buffer. We characterized 49 of 52 stool specimens from Cuban patients with amoebiosis. Among them, 36 (75.5%) were infected only with E. dispar (the nonpathogenic species), while 13 (24.5%) displayed a mixed infection with both E. dispar and E. histolytica. The multiplex PCR protocol showed a specificity of 1.00 and a sensitivity of 0.94. Furthermore, the entire procedure can be performed in one day. This approach is therefore reliable and applicable in the field for epidemiologic studies.
authors.affiliation: Institute of Tropical Medicine Pedro Kourí, Havana, Cuba.
authors.forename: Y O
authors.initials: YO
authors.lastname: Núñez
authors.forename: M A
authors.initials: MA
authors.lastname: Fernández
authors.forename: D
authors.initials: D
authors.lastname: Torres-Núñez
authors.forename: J A
authors.initials: JA
authors.lastname: Silva
authors.forename: I
authors.initials: I
authors.lastname: Montano
authors.forename: J L
authors.initials: JL
authors.lastname: Maestre
authors.forename: L
authors.initials: L
authors.lastname: Fonte
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D016054:DNA, Protozoan / Q000302:isolation & purification*; D004747:Entamoeba / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D005243:Feces / Q000469:parasitology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D016054:DNA, Protozoan