Pyrolysis mass spectrometry (PyMS) was used for rapid interstrain comparison of Pseudomonas aeruginosa isolates from a small outbreak of sternal wound infections in a cardiothoracic surgical ward. The PyMS results were compared with those obtained by conventional O-serotyping. All the isolates were phage non-typable. Evidence obtained from PyMS of identity/non-identity between isolates correlated completely with that obtained by O-typing and correctly identified those isolates comprising the epidemic strain, in one instance in advance of O-serotyping. The speed and versatility of PyMS make it an attractive technique for the initial screening of isolates from nosocomial outbreaks of infection with P. aeruginosa and other organisms.