Development and validation of a meat juice ELISA for the diagnosis of Fasciola hepatica in cattle in Cuba

ObjectiveTo establish and validate a home-made ELISA for determination of antibodies against excretory-secretory proteins of Fasciola hepatica in bovine meat juice samples.MethodsThe validity criteria of the assay were defined based on standards of International Organization for Standardization. The following parameters were evaluated: excretion/secretion antigen concentrations for coating, anti-bovine immunoglobulin G dilution, linearity, accuracy and precision.ResultsThe assay was validated on 126 meat juice samples with known infection status. Using the receiver operating characteristic (n = 126) the optimal cut-off for the ELISA assay was 0.78, above this value the probability for an animal to have fasciolosis was 11 times. And the specificity and sensitivity were 100% and 90.91% respectively. The repeatability of the intra- and inter-assay tests had coefficients of variation lower than 10% and 20% respectively.ConclusionsThe ELISA is a suitable test for further use in studies towards the epidemiology of Fasciola hepatica in Cuba.