Staphylococcus argenteus (S. argenteus) is a novel species of coagulase-positive staphylococci described in 2015. This species is phenotypically highly similar and genetically closely related to Staphylococcus aureus (S. aureus). Until recently, differentiation was only possible by molecular genetic methods, multilocus sequence typing and whole-genome sequencing, which are not generally used in routine laboratories due to time-consumingness and expensiveness. A major improvement in the identification of S. argenteus is the application of MALDI-TOF MS, if the available updated mass spectrum reference database is used. In the short report, we would like to present the first confirmed S. argenteus strain isolated from a patient in the Czech Republic and probably the first published S. argenteus strain in Central and Eastern Europe.