Determination of an optimal ELISA cut-off for the diagnosis of Toxoplasma gondii infection in pigs using Bayesian latent class modelling of data from multiple diagnostic tests

Toxoplasma gondii infection in pigs is commonly diagnosed using serological tests that detect IgG antibodies targeted against the parasite. Such tests include enzyme-linked immunosorbent assay (ELISA), modified agglutination test (MAT), and western blot (WB), which are commercially available as rapid test kits. In this study, we evaluated the manufacturer recommended cut-off of ELISA-PrioCHECK test kit and determined a new optimal cut-off for identifying T. gondii infections in pigs. Assessment of the commercial ELISA kit was done by including data from two additional serological tests, MAT, and WB, applied to seven pig population categories with varying prevalences. A total of 233 plasma samples that were previously used in other studies for investigating T. gondii seroprevalence in pigs in Denmark were randomly selected for inclusion, including 95 samples that had previously been analysed with all three tests and an additional 138 samples that were analysed using the three serological tests for this study. In the absence of a gold standard test, a latent class model was fit to the data to obtain estimates of sensitivity and specificity for each of the tests along with prevalence in each of the populations. A cut-off that maximized the sensitivity and specificity of the ELISA test was then selected. The optimal cut-off value for percent of positive control (PP) in ELISA-PrioCHECK was estimated to be 27.7 PP, which is higher than the cut-off value of 20 PP that is recommended by the manufacturer. At this cut-off, the estimated sensitivities of ELISA, MAT and WB were 99.2% (96.3-100.0%), 96.3% (88.0-100.0%), and 89.8% (80.0-98.0%), respectively. The estimated specificities of ELISA, MAT and WB were 95.2% (92.5-97.6%), 99.6% (97.5-100.0%), and 98.2% (95.9-100.0%), respectively. Our findings have broad relevance to the use of the ELISA-PrioCHECK test kit for detecting Toxoplasma gondii infection in pigs.