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

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.

Diagnosis of visceral and cutaneous leishmaniasis using loop-mediated isothermal amplification (LAMP) protocols: a systematic review and meta-analysis

“…Six datasets (IV, VI, VII, X-XII), part of three studies [ 118 , 119 , 121 ], compare LAMP to PCR for investigation of canine leishmaniasis (CL and VL) in a total of 279 animals. Datasets IV, VI and XII report an S of 100% (95% CI 95–100%), 75% (95% CI 51–91%) and 91% (95% CI 59–100%), and an E of 91% (95% CI 77–98%), 78% (95% CI 65–88%) and 96% (95% CI 86–100%), respectively.…”

Clinical Significance of ROMs, OXY, SHp and HMGB-1 in Canine Leishmaniosis

“…The reading was performed by the same observer (AP) using a fluorescence microscope. Positive and negative controls provided by CReNaL were added for each test to verify the validity of the results [ 34 ].…”

Examining the Relationship of Clinical and Laboratory Parameters With Infectiousness to Phlebotomus perniciosus and Its Potential Infectivity in Dogs With Overt Clinical Leishmaniasis

“…In each amplification run, one positive and one negative control (without DNA), both supplied by the kit, was used. Specific primer were used to amplify the 18S small subunit rRNA gene, plus a partial sequence of the internal transcribed spacer 1 (ITS-1) ( 19 ).…”