A real-time PCR assay for detection and quantification of Leishmania (Viannia) in skin and mucosal lesions: an exploratory study of parasite load and clinical parameters

M. Jara, V. Adaui, B.M. Valencia, D. Martinez Medina, M. Alba, C. Castrillon, M. Cruz, I. Cruz, G. Van der Auwera, A. Llanos-Cuentas, J.C. Dujardin, J. Arevalo

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Abstract

Earlier histopathology studies suggest that parasite load may differ between cutaneous leishmaniasis (CL) and mucosal leishmaniasis (ML) lesions, and between acute and chronic CL. Formal demonstration requires highly sensitive detection and accurate quantification of Leishmania in human lesional tissue. In this study, we developed a quantitative real-time PCR (qPCR) assay targeting the minicircle kinetoplast DNA (kDNA) to detect and quantify Leishmania (Viannia) parasites. We evaluated a total of 156 lesion biopsies from CL or ML suspected cases, and compared the quantitative performance of our kDNA-qPCR assay with a previously validated qPCR assay based on the glucose-6-phosphate dehydrogenase (G6PD) gene. We also examined the relationship between parasite load and clinical parameters. The kDNA-qPCR sensitivity for Leishmania detection was 97.9% and its specificity was 87.5%. The parasite loads quantified by kDNA-qPCR and G6PD-qPCR assays were highly correlated (r=0.87, P
Original languageEnglish
JournalJournal of Clinical Microbiology
Volume51
Issue number6
Pages (from-to)1826-1833
Number of pages8
ISSN0095-1137
DOIs
Publication statusPublished - 2013

Keywords

  • Protozoal diseases
  • Kala azar
  • Visceral
  • Cutaneous
  • Leishmaniasis
  • Leishmania donovani
  • Vectors
  • Sandflies
  • Phlebotomus argentipes
  • Detection
  • Quantification
  • Skin lesions
  • Mucosal
  • Parasite density
  • Clinical diagnosis
  • Real-time
  • Polymerase chain reaction
  • PCR
  • Assays
  • Biopsy
  • Kinetoplast DNA
  • Specificity
  • Sensitivity
  • Laboratory techniques and procedures

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