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Genus-specific kinetoplast-DNA PCR and parasite culture for the diagnosis of localised cutaneous leishmaniasis: applications for clinical trials under field conditions in Brazil.

Abstract
The positivities of two methods for the diagnosis of localised cutaneous leishmaniasis (CL) were estimated in 280 patients enrolled in a clinical trial. The trial was conducted in an endemic area of Leishmania (Viannia) braziliensis and trial participants were patients with skin ulcers and positive leishmanin skin tests. Patients underwent aspirative skin punctures of the ulcerated lesions and lymph nodes for in vitro cultures, which were processed under field conditions at the local health centre. Skin lesion biopsies were tested at a reference laboratory using kinetoplastid DNA (kDNA)-PCR to detect DNA. The median time required to obtain a positive culture from the skin samples was seven days and the contamination rate of the samples was 1.8%. The positivities of the cultures from skin lesions, kDNA-PCR and the combination of the two methods were 78.2% (95% CI: 73-82.6%), 89.3% (95% CI: 85.1-92.4%) and 97.1% (95% CI: 94.5-98.5%). We conclude that parasite culture is a feasible method for the detection of Leishmania in field conditions and that the combination of culture and PCR has a potential role for the diagnosis of CL in candidates for clinical trials.
AuthorsJulia Ampuero, Alexandre Pereira Rios, César Omar Carranza-Tamayo, Gustavo Adolfo Sierra Romero
JournalMemorias do Instituto Oswaldo Cruz (Mem Inst Oswaldo Cruz) Vol. 104 Issue 7 Pg. 992-7 (Nov 2009) ISSN: 1678-8060 [Electronic] Brazil
PMID20027466 (Publication Type: Clinical Trial, Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • DNA, Kinetoplast
Topics
  • Adolescent
  • Adult
  • Brazil
  • Cell Culture Techniques (methods)
  • Chi-Square Distribution
  • Child
  • DNA, Kinetoplast (genetics)
  • Female
  • Humans
  • Leishmania braziliensis (genetics, isolation & purification)
  • Leishmaniasis, Cutaneous (diagnosis)
  • Lymph Nodes (parasitology)
  • Male
  • Middle Aged
  • Polymerase Chain Reaction (methods)
  • Sequence Analysis, DNA
  • Skin Ulcer (parasitology)
  • Young Adult

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