Twenty-one quarters of seven cows were experimentally infected with Staphylococcus aureus (ATCC 29740) to study the shedding pattern in quarter milk samples. Of 991 consecutive quarter milk samples collected from infected quarters during the trial, 745 were positive for S. aureus by bacteriological culture. The sensitivity of a single quarter milk sample to determine infection status of a quarter was 74.5% based on the mean of each gland's recovery pattern. Sensitivity of bacterial culture increased to 94% and 98% by including a second and a third consecutive sample. Because S. aureus is shed in a cyclical manner from mammary glands, consecutive samples would be advisable for accurate diagnosis of infected quarters.