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Infection in humans has usually been related to the consumption of raw, undercooked or cured meat. The aim of this study was to develop a droplet digital polymerase chain reaction (ddPCR)-based assay for the detection and quantification of <jats:italic>T. gondii<\/jats:italic> in meat samples. To optimize the ddPCR, <jats:italic>T.gondii<\/jats:italic> reference DNA aliquots at five known concentrations: 8000 cg\/\u00b5l, 800 cg\/\u00b5l, 80 cg\/\u00b5l, 8 cg\/\u00b5l were used. Moreover, results obtained by ddPCR and quantitative PCR (qPCR) were compared using 80 known samples (40 positive and 40 negative), as well as 171 unknown diaphragm tissue samples collected at slaughterhouses. The ddPCR showed a sensitivity of 97.5% and a specificity of 100%, with a detection limit of 8 genomic copy\/\u00b5l of <jats:italic>T. gondii<\/jats:italic>. A nearly perfect agreement (<jats:italic>\u03ba<\/jats:italic>\u2009=\u20090.85) was found between results obtained by ddPCR and qPCR for both positive and negative known samples analysed. On the 171 diaphragm tissue samples from field, 7.6% resulted positive by ddPCR and only 1.2% by qPCR. 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