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Standardization and evaluation of the LAMP technique for the diagnosis of canine visceral leishmaniasis in conjunctival swab samples using DNA extracted by a silica column and boiling
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Standardization and evaluation of the LAMP technique for the diagnosis of canine visceral leishmaniasis in conjunctival swab samples using DNA extracted by a silica column and boiling
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Standardization and evaluation of the LAMP technique for the diagnosis of canine visceral leishmaniasis in conjunctival swab samples using DNA extracted by a silica column and boiling
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Journal Article
Standardization and evaluation of the LAMP technique for the diagnosis of canine visceral leishmaniasis in conjunctival swab samples using DNA extracted by a silica column and boiling
2024
Overview
The diagnosis of canine visceral leishmaniasis (CVL) presents a challenge due to a variety of non-specific clinical signs. The available tests have low sensitivity. This study aimed to standardize and evaluate the loop-mediated isothermal amplification technique with K26 target (K26-LAMP) for diagnosis of CVL in conjunctival swab (CS) DNA samples extracted through a silica column commercial kit (SW-kit) and boiling (SW-DB) and to compare sensitivity with conventional PCR (kDNA-cPCR) and quantitative real-time PCR (18S-qPCR). Clinical samples of CSs were collected from 54 dogs after reactive serology tests. Positive parasitological and/or histological tests were used as inclusion criteria for a sensitivity analysis. A total of 79.2% (43/54) of dogs without clinical signs or with mild, moderate, or severe clinical signs were included in the study. The sensitivity results of K26-LAMP, kDNA-cPCR, and 18S-qPCR were 72.1%, 81.4%, and 80.5% with the SW-kit and 97.2%, 95.2%, and 57.1% with SW-DB, respectively. In all techniques, the proportion of positives was higher in the group with severe clinical disease, with statistically significant differences in the K26-LAMP and 18S-qPCR techniques being seen with the SW-kit. The results obtained with LAMP for CS samples are promising and its performance is similar to other techniques.
Publisher
MDPI,MDPI AG
Subject
/ Animals
/ canine visceral leishmaniasis
/ DNA
/ Dogs
/ Ethylenediaminetetraacetic acid
/ Methods
/ Protozoa
/ Silica
/ Spleen
