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In human medicine, procalcitonin (PCT) is accepted as a highly specific and early marker for microbial infections and sepsis. The aim of this study was to compare plasma PCT concentrations in horses questionable of sepsis to healthy individuals using a newly validated sandwich-ELISA for the detection of equine PCT. Plasma samples of 24 healthy horses and five horses with clinical signs of sepsis were tested. All horses were classified according to a sepsisscore adapted from Breuer et al. (2012) including data from clinical examination and laboratory data at the time of first presentation as positive for sepsis (group A, ≥ 7 score points), questionable (group B, 4–6 score points), or negative for sepsis (group C, ≤ 3 score points). The five horses with clinical signs of sepsis were assigned to group A (2 mares, 3 geldings, age: 15 4 years, bodyweight: 520 68 kg), 4 healthy horses (3 geldings and 1 mare, age: 14 5 years, bodyweight: 575 86 kg) were classified as group B and 20 healthy horses (14 geldings and 6 mares; age: 13 5 years, BDW: 529 58 kg) were classified as group C. We found a significantly higher median ePCT concentration of 8450 ng/mL in the sepsis group in contrast to 47 ng/mL for the control groups B and C (P = 0.0006). All plasma samples of the sepsis group showed increased ePCT indicating the relevance of ePCT as a valuable sepsis marker in horses.