Quantification of Leishmania infantum parasites in tissue biopsies by real-time polymerase chain reaction and polymerase chain reaction-enzyme-linked immunosorbent assay

N Rolão, S Cortes, O R Rodrigues, L Campino

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

Most of the experimental studies of Leishmania spp. infection require the determination of the parasite load in different tissues. Quantification of parasites by microscopy is not very sensitive and is time consuming, whereas culture microtitrations remain laborious and can be jeopardized by microbial contamination. The aim of this study was to quantify Leishmania infantum parasites by real-time polymerase chain reaction (PCR) using specific DNA TaqMan probes and to compare the efficacy of detection of this technique with a PCR-enzyme-linked immunosorbent assay (ELISA). For this purpose, spleen and liver samples from L. infantum-infected mice were collected during a 3-mo longitudinal study and analyzed by both methods. PCR-ELISA failed to quantify Leishmania spp. DNA in samples with very low or very high numbers of parasites. Real-time PCR was more sensitive than PCR-ELISA, detecting down to a single parasite, and enabled the parasite quantification over a wide, 5-log range. In summary, this study developed a method for absolute quantification of L. infantum parasites in infected organs using real-time TaqMan PCR.

Original languageEnglish
Pages (from-to)1150-1154
Number of pages5
JournalJournal Of Parasitology
VolumeVol. 90
Issue numbern.º 5
DOIs
Publication statusPublished - 1 Oct 2004

Keywords

  • Animals
  • Biopsy
  • DNA, Protozoan
  • Disease Models, Animal
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Leishmania infantum
  • Leishmaniasis, Visceral
  • Liver
  • Longitudinal Studies
  • Mice
  • Polymerase Chain Reaction
  • Reproducibility of Results
  • Spleen
  • Comparative Study
  • Journal Article
  • Research Support, Non-U.S. Gov't

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