Comparative Study of Real-Time PCR (TaqMan Probe and Sybr Green), Serological Techniques (ELISA, IFA and DAT) and Clinical Signs Evaluation, for the Diagnosis of Canine Leishmaniasis in Experimentally Infected Dogs.

María Paz Peris,Adriana Esteban-Gil,Juan Antonio Castillo,Nabil Halaihel,Mariano Morales,Paula Ortega-Hernández

doi:10.3390/microorganisms9122627

María Paz Peris, Adriana Esteban-Gil + Show 4 more

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https://doi.org/10.3390/microorganisms9122627

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Journal: Microorganisms	Publication Date: Dec 20, 2021
Citations: 8	License type: CC BY 4.0

Affiliation: Universidad de Zaragoza

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Canine leishmaniasis (CanL) diagnosis is not fully resolved. Currently, two specific methodologies are in continuous development, the detection of the parasite DNA or RNA in target organs and the detection of specific antibodies against Leishmania sp. For a correct diagnosis, it has been shown that the joint use of this type of test is necessary. In this work, a Sybr Green and a TaqMan Probe based on real time PCRs (qPCR) was performed for the detection of Leishmania sp. in order to correlate the results with clinicopathological and serological evaluations (IFA, ELISA and DAT) to propose an optimal biological sample to be used to detect the parasite in both early and late stages of the infection. A total of four samples were processed: conjunctival swabs, popliteal lymph node aspirates, bone marrow aspirates, and peripheral blood from experimentally infected dogs belonging to a larger study. Our results indicated that a single non-invasive sample (conjunctival swab) and the application of both types of qPCR would be reliable for determining Leishmania infection as well as the disease stage in dogs, thus avoiding bone marrow, lymph node aspirate or blood samples collection.

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The parasite Leishmania infantum has been identified as the main etiologic agent of canine visceral leishmaniasis, (CVL) which is a major global zoonosis that is potentially fatal to humans and dogs [2]
In recent studies where the skin, lymph node, bone marrow and conjunctival swab were evaluated, they concluded that the skin [20] and the conjunctival swab [21] were the tissues that showed the highest rate of positivity
The highest parasite load corresponded to the bone marrow, followed by the lymph node, conjunctival swabs and the peripheral blood

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Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Leishmaniasis is one of the most important neglected tropical diseases according to WHO [1]. The parasite Leishmania infantum has been identified as the main etiologic agent of canine visceral leishmaniasis, (CVL) which is a major global zoonosis that is potentially fatal to humans and dogs [2]. Several organs can be affected during the progression of the disease, including the skin, kidneys, spleen, liver, and eyes, and is characterized by a range of associated clinical signs such as skin lesions, generalized lymphadenopathy, weight loss, muscle atrophy, intolerance to exercise, loss of appetite, lethargy, splenomegaly, polyuria, polydipsia, ocular lesion, epistaxis, onychogryphosis, anemia, vomiting, and diarrhea [3]

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