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      Clinical, Molecular and Serological Diagnosis of Canine Leishmaniosis: An Integrated Approach

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          Abstract

          Canine leishmaniosis (CanL) is caused by protozoans of the genus Leishmania and characterized by a broad spectrum of clinical signs in dogs. Early diagnosis is of great importance in order to perform an appropriate therapy and to prevent progression towards severe disease. The aim of this study was to compare a point-of-care molecular technique, i.e., the loop-mediated isothermal amplification (LAMP), with a real-time polymerase chain reaction (Rt-PCR), and three serological techniques, i.e., immunofluorescence antibody test (IFAT), enzyme-linked immunosorbent assay (ELISA), and a rapid SNAP Leishmania test, to develop an integrated approach for the diagnosis of CanL. Sixty dogs were chosen after physical examination and collection of blood and sera samples, fine-needle aspiration of lymph nodes, and conjunctival swabs were performed. Lymphadenopathy (82.3%), as well as clinicopathological alterations of total proteins (70.6%), were the most frequent signs. Forty-one (68.3%) samples resulted positive at least to one technique. IFAT resulted in the best serological diagnostic method (specificity = 100%, sensitivity = 97.2%), detecting a higher number of positive samples than those revealed by other techniques. Among the samples used for molecular analysis, fine-needle aspiration of lymph nodes was revealed as the best sample source. LAMP showed a substantial agreement ( κ = 0.80; p <0.0001) with Rt-PCR; therefore, it could be promising for the rapid diagnosis of CanL. Nevertheless, further studies should be performed to confirm these findings.

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          LeishVet guidelines for the practical management of canine leishmaniosis

          The LeishVet group has formed recommendations designed primarily to help the veterinary clinician in the management of canine leishmaniosis. The complexity of this zoonotic infection and the wide range of its clinical manifestations, from inapparent infection to severe disease, make the management of canine leishmaniosis challenging. The recommendations were constructed by combining a comprehensive review of evidence-based studies, extensive clinical experience and critical consensus opinion discussions. The guidelines presented here in a short version with graphical topic displays suggest standardized and rational approaches to the diagnosis, treatment, follow-up, control and prevention of canine leishmaniosis. A staging system that divides the disease into four stages is aimed at assisting the clinician in determining the appropriate therapy, forecasting prognosis, and implementing follow-up steps required for the management of the leishmaniosis patient.
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            Canine leishmaniasis: epidemiological risk and the experimental model.

            Increasing risk factors are making zoonotic visceral leishmaniasis a growing public health concern in many countries. Domestic dogs constitute the main reservoir of Leishmania infantum and Leishmania chagasi, and play a key role in the transmission to humans. New reagents and tools allow the detailed investigation of canine leishmaniasis, permitting the monitoring of the immunological status of dogs in both natural and experimental infections. Such studies are essential to determine the basis of the canine protective immune response and to establish a laboratory model, a significant aspect for the development of vaccines against canine leishmaniasis.
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              A review of solutions for diagnostic accuracy studies with an imperfect or missing reference standard.

              In diagnostic accuracy studies, the reference standard may be imperfect or not available in all patients. We systematically reviewed the proposed solutions for these situations and generated methodological guidance. Review of methodological articles. We categorized the solutions into four main groups. The first group includes methods that impute or adjust for missing data on the reference standard. The second group consists of methods that correct estimates of accuracy obtained with an imperfect reference standard. In the third group a reference standard is constructed by combining multiple test results through a predefined rule, based on a consensus procedure, or through statistical modeling. In the fourth group, the diagnostic accuracy paradigm is abandoned in favor of validation studies that relate index test results to relevant clinical data, such as history, future clinical events, and response to therapy. Most of the methods try to impute, adjust, or construct a reference standard. In situations that deviate only marginally from the classical diagnostic accuracy paradigm, these are valuable methods. In cases where an acceptable reference standard does not exist, the concept of clinical test validation may provide an alternative paradigm to evaluate a diagnostic test.
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                Author and article information

                Journal
                Vet Sci
                Vet Sci
                vetsci
                Veterinary Sciences
                MDPI
                2306-7381
                14 April 2020
                June 2020
                : 7
                : 2
                : 43
                Affiliations
                [1 ]Department of Veterinary Medicine and Animal Production, University of Naples Federico II, 80137 Naples, Italy; mariapaola.maurelli@ 123456unina.it (M.P.M.); boscoant@ 123456tiscali.it (A.B.); daniela.giaquinto@ 123456gmail.com (D.G.); lavinia_vet1@ 123456yahoo.com (L.C.); giuseppe.molinaro@ 123456unina.it (G.M.); giuseppe.cringoli@ 123456unina.it (G.C.); gaetano.oliva@ 123456unina.it (G.O.); laura.rinaldi@ 123456unina.it (L.R.); manuela.gizzarelli@ 123456unina.it (M.G.)
                [2 ]Regional Center for Monitoring Parasitic Diseases (CREMOPAR), Campania Region, 84025 Eboli (Sa), Italy
                [3 ]National Reference Center for Leishmaniosis, Istituto Zooprofilattico Sperimentale della Sicilia, 90129 Palermo, Italy; fabrizio.vitale@ 123456izssicilia.it
                Author notes
                Author information
                https://orcid.org/0000-0002-7564-3356
                Article
                vetsci-07-00043
                10.3390/vetsci7020043
                7378766
                32295198
                954bd31e-02c4-42b8-961b-7a00fb99ee30
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 21 March 2020
                : 10 April 2020
                Categories
                Article

                leishmaniosis,dog,loop-mediated isothermal amplification (lamp),real-time polymerase chain reaction (rt-pcr),immunofluorescence antibody test (ifat),enzyme-linked immunosorbent assay (elisa),snap leishmania test

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