The study describes the use of xMAP technology for in vitro multiplex detection and differentiation of bacterial agents capable of inducing abscess formation in small ruminants, with a particular focus on Corynebacterium pseudotuberculosis causing caseous lymphadenitis in sheep and goats. Functionality of the diagnostic panel for Corynebacterium pseudotuberculosis, Corynebacterium ulcerans, Staphylococcus aureus subsp. aureus, Staphylococcus aureus subsp. anaerobius, Streptococcus spp., Trueperella pyogenes and methicillin-resistant Staphylococcus aureus was tested on reference strains from the Collection of Animal Pathogenic Microorganisms at the Veterinary Research Institute in Brno and using lysates from field bacterial isolates typed by MALDI-TOF mass spectrometry. The detection system was subsequently used to test real pus samples from abscesses of sheep and goats collected in the Czech Republic. xMAP technology is a suitable diagnostic tool especially as a supplementation to culture and typing methods used in routine diagnostic practice.