title: Simultaneous detection and quantification of Ancylostoma duodenale, Necator americanus, and Oesophagostomum bifurcum in fecal samples using multiplex real-time PCR.
abstract: A multiplex real-time PCR was developed and evaluated for the simultaneous detection of Ancylostoma duodenale, Necator americanus, and Oesophagostomum bifurcum in fecal samples. Using well-defined control samples (N = 150), known positive fecal samples (N = 50), and fecal samples from an area in Ghana where human infections with all 3 nematode species are endemic (N = 339), the method proved to be highly specific and sensitive. Cycle threshold (Ct) values, reflecting parasite-specific DNA load, showed significant correlation with the intensity of infection as measured by microscopy using Kato-Katz fecal smears or by species specific third-stage larval count after coproculture. The multiplex real-time PCR described combined with the simple fecal sample collection procedure and the potential for high throughput makes this approach a powerful diagnostic tool to study species-specific transmission patterns of human hookworm-like infections. Moreover, this procedure facilitates monitoring of intervention programs and allows species-specific detection of treatment failure following rounds of mass treatment.
authors.affiliation: Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands. j.j.verweij@lumc.nl
authors.forename: Jaco J
authors.initials: JJ
authors.lastname: Verweij
authors.forename: Eric A T
authors.initials: EA
authors.lastname: Brienen
authors.forename: Juventus
authors.initials: J
authors.lastname: Ziem
authors.forename: Lawrence
authors.initials: L
authors.lastname: Yelifari
authors.forename: Anton M
authors.initials: AM
authors.lastname: Polderman
authors.forename: Lisette
authors.initials: L
authors.lastname: Van Lieshout
mesh_terms: D000722:Ancylostoma / Q000235:genetics*; D000724:Ancylostomiasis / Q000175:diagnosis / Q000469:parasitology*; D000818:Animals; D005243:Feces / Q000469:parasitology; D006801:Humans; D017221:Necator americanus / Q000235:genetics*; D009332:Necatoriasis / Q000175:diagnosis / Q000469:parasitology*; D009814:Oesophagostomiasis / Q000175:diagnosis / Q000469:parasitology*; D009815:Oesophagostomum / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity