title: Rapid identification of Leishmania complexes by a real-time PCR assay.
abstract: A real-time PCR assay for the detection of four Leishmania complexes (L. Viannia, L. mexicana, L. donovani/infantum, and L. major) was developed and evaluated. The assay was developed to detect the glucosephosphate isomerase gene and capitalizes on DNA sequence variability within that gene for Leishmania complex identification. Primer/probe sets were created and tested against a panel of 21 known negative controls and on DNA extracted from cultured promastigotes or from tissue biopsies from patients with cutaneous leishmaniasis. The assay was highly specific, as no amplification products were detected in the negative control samples while simultaneously retaining a high degree of complex-specific diagnostic accuracy for cultured organisms and patient clinical samples. Real-time PCR offers rapid (within hours) identification of Leishmania to the complex level and provides a useful molecular tool to assist both epidemiologists and clinicians.
authors.affiliation: Infectious Disease Service, Walter Reed Army Medical Center, Washington, DC 20307-5001, USA. glenn.wortmann@na.amedd.army.mil
authors.forename: Glenn
authors.initials: G
authors.lastname: Wortmann
authors.forename: Lisa
authors.initials: L
authors.lastname: Hochberg
authors.forename: Huo-Hsu
authors.initials: HH
authors.lastname: Houng
authors.forename: Colleen
authors.initials: C
authors.lastname: Sweeney
authors.forename: Michael
authors.initials: M
authors.lastname: Zapor
authors.forename: Naomi
authors.initials: N
authors.lastname: Aronson
authors.forename: Peter
authors.initials: P
authors.lastname: Weina
authors.forename: Christian F
authors.initials: CF
authors.lastname: Ockenhouse
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D016054:DNA, Protozoan / Q000032:analysis; D005956:Glucose-6-Phosphate Isomerase / Q000235:genetics*; D006801:Humans; D007891:Leishmania / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis* / Q000469:parasitology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D016415:Sequence Alignment
chemical_list: D017931:DNA Primers; D016054:DNA, Protozoan; D011933:Reagent Kits, Diagnostic; D005956:Glucose-6-Phosphate Isomerase