Detection and identification of Kinetoplastids of zoonotic interest by HRM-qPCR analysis in Canis lupus familiaris from Argentinean Mesopotamia

• Published in: Veterinary parasitology (Amsterdam) Vol. 24; p. 100557
• Main Authors: Muñoz-Calderón, Arturo, Lucero, Raul Horacio, Brusés, Bettina L., Formichelli, Laura, Koscinczuk, Patricia, Pedelhez, Mariana, Schijman, Alejandro G.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.04.2021
• Subjects: Animals; Canis lupus familiaris; Dogs; Leishmania - genetics; Leishmania spp; Mesopotamia; Molecular diagnosis; Real-time PCR; Real-Time Polymerase Chain Reaction - methods; Real-Time Polymerase Chain Reaction - veterinary; Trypanosoma cruzi - genetics; Trypanosoma evansi; Wolves; Zoonosis; Mesopotamia; Trypanosoma evansi; Molecular diagnosis; Zoonosis; Real-time PCR; Canis lupus familiaris; Leishmania spp
• ISSN: 2405-9390; 2405-9390
• DOI: 10.1016/j.vprsr.2021.100557

This work aimed to conduct a first PCR-based approach for differential diagnosis of kinetoplastidean infections in dogs. Diagnosis of Kinetoplastid infections in domestic animals is difficult, since parasitemia is intermittent and signs are nonspecific; it is mainly based on parasitological smears or concentration techniques, which lack sensitivity and depend on operator` expertise. Dogs are relevant reservoirs in transmission of Kinetoplastids; they function as sentinels to detect active transmission cycles before they involve humans. Trypanosoma cruzi, Trypanosoma evansi, and various species of Leishmania genus are multi-host parasites, capable of parasitizing dogs among a vast number of reservoirs. An algorithm based on sequential Real-Time PCR-High Resolution Melting (HRM) (qPCR-HRM) assays directed at 24S alpha ribosomal DNA, ITS1 and Hsp70 designed to distinguish among T. cruzi, T. rangeli, T. evansi and Leishmania spp. was tested in fourteen dogs with suspicion of kinetoplastid diseases. A qPCR control of DNA integrity in the tested sample, targeted to the mammalian interphotoreceptor retinoid-binding protein (IRBP) gene fragment was incorporated to the algorithm. T. evansi was detected in four dogs and L. infantum in one. Two of five qPCR positive cases were smear negative. Smear and T. evansi qPCR positive cases corresponded to animals that died despite being treated, indicating the association of parasitemia with disease severity. This laboratory tool increases the possibility of confirming outbreaks of kinetoplastid diseases with zoonotic potential and identify the etiological agents involved. •A qPCR-HRM algorithm allowed differential detection of trypanomicidal infections in domestic dogs.•qPCR was higher sensitive than smear to detect these infections.•Native Trypanosoma evansi infections occur in domestic dogs in Argentinean Mesopotamia•Leishmania infantum infections were also detected in dogs at the same localities.•Smear plus T. evansi qPCR positivity were associated to disease severity.