Dientamoeba fragilis diagnosis by fecal screening: relative effectiveness of traditional techniques and molecular methods

Abstract

Introduction: Dientamoeba fragilis, an intestinal trichomonad, occurs in humans with and without gastrointestinal symptoms. Its presence was investigated in individuals referred to Milad Hospital, Tehran. Methodology: In a cross-sectional study, three time-separated fecal samples were collected from 200 participants from March through June 2011. Specimens were examined using traditional techniques for detecting D. fragilis and other gastrointestinal parasites: direct smear, culture, formalin-ether concentration, and iron-hematoxylin staining. The presence of D. fragilis was determined using PCR assays targeting 5.8S rRNA or small subunit ribosomal RNA. Results: Dientamoeba fragilis, Blastocystis sp., Giardia lamblia, Entamoeba coli, and Iodamoeba butschlii were detected by one or more traditional and molecular methods, with an overall prevalence of 56.5%. Dientamoeba was not detected by direct smear or formalin-ether concentration but was identified in 1% and 5% of cases by culture and iron-hematoxylin staining, respectively. PCR amplification of SSU rRNA and 5.8S rRNA genes diagnosed D. fragilis in 6% and 13.5%, respectively. Prevalence of D. fragilis was unrelated to participant gender, age, or gastrointestinal symptoms. Conclusions: This is the first report of molecular assays to screen for D. fragilis in Iran. The frequent finding of D. fragilis via fecal analysis indicated the need to include this parasite in routine stool examination in diagnostic laboratories. As the length of amplification target correlates to the sensitivity of PCR, this assay targeting the D. fragilis 5.8S rRNA gene seems optimal for parasite detection and is recommended in combination with conventional microscopy for diagnosing intestinal parasites.