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Background  Canine leishmaniasis, a zoonotic disease caused by  Leishmania infantum  vectored by phlebotomine sand flies, is considered a relevant veterinary and public health problem in various countries, namely in the Mediterranean basin and Brazil, where dogs are considered the main reservoir hosts. Not only diseased dogs but also those subclinically infected play a relevant role in the transmission of  L. infantum  to vectors; therefore, early diagnosis is essential, under both a clinical and an epidemiological perspective. Molecular tools can be a more accurate and sensitive approach for diagnosis, with a wide range of protocols currently in use. The aim of the present report was to compare four PCR based protocols for the diagnosis of canine  Leishmania  infection in a cohort of dogs from the Douro region, Portugal.    Results  A total of 229 bone marrow samples were collected from dogs living in the Douro region, an endemic region for leishmaniasis. Four PCR protocols were evaluated for  Leishmania  DNA detection in canine samples, three single (ITS1-PCR, MC-PCR and Uni21/Lmj4-PCR) and one nested (nested SSU rRNA-PCR). Two of the protocols were based on nuclear targets and the other two on kinetoplastid targets. The higher overall percentage of infected dogs was detected with the nested SSU rRNA-PCR (37.6%), which also was able to detect  Leishmania  DNA in a higher number of samples from apparently healthy dogs (25.3%). The ITS1-PCR presented the lowest level of  Leishmania  detection.    Conclusions  Nested SSU rRNA-PCR is an appropriate method to detect  Leishmania  infection in dogs. Accurate and early diagnosis in clinically suspect as well as apparently healthy dogs is essential, in order to treat and protect animals and public health and contribute to the control and awareness of the disease.

Evaluation of four molecular methods to detect Leishmania infection in dogs