Rapid recognition ofStaphylococcalbacteremia in humans is a serious challenge in clinical research. Here we report on direct analysis of molecular volatile organic compounds (VOCs) of blood cultures emitted by 35 patients with a suspicion ofStaphylococcalbacteremia and 31 patients positively diagnosed withStaphylococcalbacteremia were fingerprinted by atmospheric corona discharge ionization mass spectrometry (CDI-MS) for the presence of four commonStaphylococci(Staphylococcus aureus,S. hominis,S. haemolyticus, andS. epidermidis) which statistically account forca.90% of cases ofStaphylococcalbacteremia in humans. All the infected blood cultures revealed characteristic and clearly distinct MS patterns specific to the presence of one of the fourStaphylococciafter an incubation time of 3–16 h. Technical replicates were incubated over 2–3 days for reference diagnosis using current conventional bacteremia detection. The results of our 6 month hospital study confirm peaks atm/z87 (isobutyric acid) andm/z101 (isovaleric acid) signals were specifically present inStaphylococci, and indicate that high suitability of CDI-MS for rapid, reliable and cost-efficient diagnosis ofStaphylococcalbacteremia in humans.