title: Application of Multiplex PCR for Detection and Differentiation of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii.
abstract: BACKGROUND
  Entamoeba moshkovskii and E. dispar are impossible to differentiate microscopically from the pathogenic species E. histolytica. Multiplex polymerase chain reaction (Multiplex PCR) is a widespread molecular biology technique for amplification of multiple targets in a single PCR experiment.
  
  
  METHODS
  For detection and differentiation of the three-microscopy indistinguishable Entamoeba species in human, multiplex PCR assay using different DNA extraction methods was studied. A conserved forward primer was derived from the middle of the small-subunit rRNA gene, and reverse primers were designed from signature sequences specific to each of these three Entamoeba species.
  
  
  RESULTS
  A 166-bp PCR product with E. histolytica DNA, a 580-bp product with E. moshkovskii DNA and a 752-bp product with E. dispar DNA were generated in a single-round and multiplex PCR reaction.
  
  
  CONCLUSIONS
  We recommend this PCR assay as an accurate, rapid, and effective diagnostic method for the detection and discrimination of these three Entamoeba species in both routine diagnosis of amoebiasis and epidemiological surveys.
authors.affiliation: Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Nozhat
authors.initials: N
authors.lastname: Zebardast
authors.affiliation: Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Haghighi
authors.affiliation: Dept. of Immunology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Farshid
authors.initials: F
authors.lastname: Yeganeh
authors.affiliation: Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Seyyed Javad
authors.initials: SJ
authors.lastname: Seyyed Tabaei
authors.affiliation: Dept. of Parasitology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran.
authors.forename: Mohammad Javad
authors.initials: MJ
authors.lastname: Gharavi
authors.affiliation: Dept. of Parasitology and Mycology, Lorestan University of Medical Sciences, Khorramabad, Iran.
authors.forename: Shirzad
authors.initials: S
authors.lastname: Fallahi
authors.affiliation: Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Zohreh
authors.initials: Z
authors.lastname: Lasjerdi
authors.affiliation: Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Nima
authors.initials: N
authors.lastname: Salehi
authors.affiliation: Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Niloofar
authors.initials: N
authors.lastname: Taghipour
authors.affiliation: Tamin Ejtemaee Hospital, Ahvaz, Iran.
authors.forename: Cobra
authors.initials: C
authors.lastname: Kohansal
authors.affiliation: Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Farideh
authors.initials: F
authors.lastname: Naderi
keywords: DNA extraction; Entamoeba dispar; Entamoeba hitolytica; Entamoeba moshkovskii; Multiplex PCR