title: Detection and differentiation of Entamoeba histolytica and Entamoeba dispar in clinical samples through PCR-denaturing gradient gel electrophoresis.
abstract: Amebiasis is one of the twenty major causes of disease in Mexico; however, the diagnosis is difficult due to limitations of conventional microscopy-based techniques. In this study, we analyzed stool samples using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) to differentiate between Entamoeba histolytica (pathogenic) and E. dispar (non-pathogenic). The target for the PCR amplification was a small region (228 bp) of the adh112 gene selected to increase the sensitivity of the test. The study involved 62 stool samples that were collected from individuals with complaints of gastrointestinal discomfort. Of the 62 samples, 10 (16.1%) were positive for E. histolytica while 52 (83.9%) were negative. No sample was positive for E. dispar. These results were validated by nested PCR-RFLP (restriction fragment length polymorphism) and suggest that PCR-DGGE is a promising tool to differentiate among Entamoeba infections, contributing to determine the specific treatment for patients infected with E. histolytica, and therefore, avoiding unnecessary treatment of patients infected with the non-pathogenic E. dispar.
authors.affiliation: Family Medicine Unit 43, Social Security Mexican Institute, Cardenas, Tabasco, Mexico.
authors.forename: P
authors.initials: P
authors.lastname: López-López
authors.affiliation: Health Sciences Academic Division, Juarez Autonomous University of Tabasco, Villahermosa, Tabasco, Mexico.
authors.forename: M C
authors.initials: MC
authors.lastname: Martínez-López
authors.affiliation: Health Sciences Academic Division, Juarez Autonomous University of Tabasco, Villahermosa, Tabasco, Mexico.
authors.forename: X M
authors.initials: XM
authors.lastname: Boldo-León
authors.affiliation: Multidisciplinary Academic Division at Jalpa de Mendez, Juarez Autonomous University of Tabasco, Jalpa de Mendez, Tabasco, Mexico.
authors.forename: Y
authors.initials: Y
authors.lastname: Hernández-Díaz
authors.affiliation: Multidisciplinary Academic Division at Jalpa de Mendez, Juarez Autonomous University of Tabasco, Jalpa de Mendez, Tabasco, Mexico.
authors.forename: T B
authors.initials: TB
authors.lastname: González-Castro
authors.affiliation: Multidisciplinary Academic Division at Comalcalco, Juarez Autonomous University of Tabasco, Comalcalco, Tabasco, Mexico.
authors.forename: C A
authors.initials: CA
authors.lastname: Tovilla-Zárate
authors.affiliation: Center for Research and Advanced Studies, National Polytechnic Institute, Mexico City, Mexico.
authors.forename: J P
authors.initials: JP
authors.lastname: Luna-Arias
mesh_terms: D016054:DNA, Protozoan / Q000235:genetics; D058645:Denaturing Gradient Gel Electrophoresis / Q000379:methods*; D004747:Entamoeba / Q000235:genetics* / Q000302:isolation & purification*; D004748:Entamoeba histolytica / Q000235:genetics* / Q000302:isolation & purification*; D004749:Entamoebiasis / Q000469:parasitology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D015203:Reproducibility of Results
chemical_list: D016054:DNA, Protozoan