Development and validation of an in-house ELISA to detect Leishmania parasites in dogs

Abstract

Introduction: Leishmaniasis, a vector-borne parasitic disease, caused by Leishmania donovani is endemic in Sri Lanka. Inconclusive evidence suggests the possibility of dogs being a reservoir for Leishmania. Identification of animal reservoirs is crucial to control the infection. The objective of this study was to develop and validate a low-cost ELISA method to screen for Leishmania in dogs. Methods: An experimental study was conducted. Leishmania parasites were cultured, and crude protein lysate was extracted and quantified. The ELISA was optimised using checkerboard titration and was validated using a commercial kit. Accuracy, precision, linearity, specificity and sensitivity of the in-house ELISA were determined. The crosstab analysis was performed to determine the sensitivity and specificity of the in-house method. Blood samples were collected from 71 dogs: 60 from Kilinochchi and 11 from Mirigama. The collected serum samples were analysed using the optimised in-house ELISA.