Microbiological contamination of air in the operating room is generally considered to be a risk factor for surgical site infections in clean surgery. Evaluation of the quality of air in operating theatres can be performed routinely by microbiological sampling and particle counting, but the relationship between these two methods has rarely been evaluated. The aim of this study was to determine whether particle counting could be predictive of microbiological contamination of air in operating rooms. Over a three-month period, air microbiological sampling and particle counting were performed simultaneously in four empty operating rooms belonging to two surgical theatres equipped with conventional ventilation via high-efficiency particulate air filters. Correlation between the two methods was measured with Spearman's correlation coefficient. The ability of particle counting to discriminate between microbiological counting values higher and lower than 5 colony-forming units (CFU)/m3 was evaluated using receiver-operating characteristic (ROC) analysis. Microbiological counting ranged from 0 to 38CFU/m3, while the particle counts ranged from 0 to 46 262/m3. Methods of microbiological and particle counting did not correlate (Spearman correlation coefficient=0.06, P=0.6). Using the ROC curve, no particle count value could be predictive of a microbiological count higher than 5CFU/m3. The results of the current study suggest that there is no reason to replace microbiological sampling with particle counting for routine evaluation of microbiological contamination in conventionally ventilated operating theatres.