Advantages of real-time PCR assay for diagnosis and monitoring of canine leishmaniosis

O. Francino, L. Altet, E. Sánchez-Robert, A. Rodriguez, L. Solano-Gallego, J. Alberola, L. Ferrer, A. Sánchez, X. Roura

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The aim of the present study is to highlight the advantages of real-time quantitative PCR intended to aid in the diagnosis and monitoring of canine leishmaniosis. Diagnosis of canine leishmaniosis is extremely challenging, especially in endemic areas, due to the diverse and non-specific clinical manifestations, and due to the high seroprevalence rate in sub-clinical dogs. Veterinarian clinicians are usually confronted with cases that are compatible with the disease, and with several diagnostic tests, sometimes with contradictory results. We have developed a new TaqMan assay, targeting the kinetoplast, applied to 44 samples of bone marrow aspirate or peripheral blood. The dynamic range of detection of Leishmania DNA was established in 7 logs and the limit of detection is 0.001 parasites in the PCR reaction. At the time of diagnosis parasitemia ranges from less than 1 to 107 parasites/ml. The ability to quantify the parasite burden allowed: (i) to elucidate the status of positive dogs by conventional PCR, although larger studies are necessary to clarify the dividing line between infection and disease, (ii) to estimate the kinetics of the parasite load and the different response to the treatment in a follow-up and (iii) to validate blood as less invasive sample for qPCR. The continuous data provided by real-time qPCR could solve the dilemma for the clinician managing cases of canine leishmaniosis by differentiating between Leishmania-infected dogs or dogs with active disease of leishmaniosis. © 2006 Elsevier B.V. All rights reserved.
Original languageEnglish
Pages (from-to)214-221
JournalVeterinary Parasitology
Issue number3-4
Publication statusPublished - 30 Apr 2006


  • Dog
  • Leishmania infantum
  • Parasite quantification
  • Real-time PCR


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