title: A simplified and standardized polymerase chain reaction format for the diagnosis of leishmaniasis.
abstract: BACKGROUND
  Definite diagnosis of Leishmania infections is based on demonstration of the parasite by microscopic analysis of tissue biopsy specimens or aspirate samples. However, microscopy generally shows low sensitivity and requires invasive sampling.
  
  
  METHODS
  We describe here the development of a simple and rapid test for the detection of polymerase chain reaction-amplified Leishmania DNA. A phase 1 evaluation of the text was conducted in clinical samples from 60 nonendemic and 45 endemic control subjects and from 44 patients with confirmed cutaneous leishmaniasis (CL), 12 with mucocutaneous leishmaniasis (MCL), and 43 with visceral leishmaniasis (VL) from Peru, Kenya, and Sudan.
  
  
  RESULTS
  The lower detection limits of the assay are 10 fg of Leishmania DNA and 1 parasite in 180 microL of blood. The specificity was 98.3% (95% confidence interval [CI], 91.1%-99.7%) and 95.6% (95% CI, 85.2%-98.8%) for nonendemic and endemic control samples, respectively, and the sensitivity was 93.2% (95% CI, 81.8%-97.7%), 91.7% (95% CI, 64.6%-98.5%), and 86% (95% CI, 72.7%-93.4%) for lesions from patients with CL or MCL and blood from patients with VL, respectively.
  
  
  CONCLUSIONS
  The Leishmania OligoC-TesT showed high specificity and sensitivity in clinical samples and was able to detect the parasite in samples obtained by less invasive means, such as blood, lymph, and lesion scrapings. The assay is a promising new tool for simplified and standardized molecular detection of Leishmania parasites.
authors.affiliation: Department of Parasitology, Institute of Tropical Medicine Antwerp, Antwerp, Belgium. sdeborggraeve@itg.be
authors.forename: Stijn
authors.initials: S
authors.lastname: Deborggraeve
authors.forename: Thierry
authors.initials: T
authors.lastname: Laurent
authors.forename: Diego
authors.initials: D
authors.lastname: Espinosa
authors.forename: Gert
authors.initials: G
authors.lastname: Van der Auwera
authors.forename: Margaret
authors.initials: M
authors.lastname: Mbuchi
authors.forename: Monique
authors.initials: M
authors.lastname: Wasunna
authors.forename: Sayda
authors.initials: S
authors.lastname: El-Safi
authors.forename: Ahmed Almustafa
authors.initials: AA
authors.lastname: Al-Basheer
authors.forename: Jorge
authors.initials: J
authors.lastname: Arévalo
authors.forename: César
authors.initials: C
authors.lastname: Miranda-Verástegui
authors.forename: Thierry
authors.initials: T
authors.lastname: Leclipteux
authors.forename: Pascal
authors.initials: P
authors.lastname: Mertens
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Dujardin
authors.forename: Piet
authors.initials: P
authors.lastname: Herdewijn
authors.forename: Philippe
authors.initials: P
authors.lastname: Büscher
mesh_terms: D000818:Animals; D001483:Base Sequence; D006801:Humans; D007891:Leishmania / Q000235:genetics* / Q000302:isolation & purification; D007896:Leishmaniasis / Q000097:blood / Q000175:diagnosis*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012680:Sensitivity and Specificity; D016415:Sequence Alignment
chemical_list: D012337:RNA, Ribosomal, 18S