To determine methicillin-resistant Staphylococcus aureus (MRSA) carriage in poultry and slaughterhouse personnel, 40 Dutch broiler flocks, in six slaughterhouses and 466 personnel were sampled. Of the employees, 26 were positive (5·6%), indicating a higher risk of exposure when compared to the general Dutch population (0·1%). This risk was significantly higher for personnel having contact with live animals (5·2%) – especially hanging broilers on the slaughterline (20·0%) – than for all other personnel (1·9%). Conventional electric stunning conferred a significantly higher risk of MRSA carriage for employees than CO2 stunning (9·7% vs. 2·0%). A total of 405 broilers were sampled upon their arrival at the slaughterhouse, of which 6·9% were positive. These broilers originated from 40 Dutch slaughter flocks of which 35·0% were positive. MRSA contamination in the different compartments of slaughterhouses increased during the production day, from 8% to 35%. Of the 119 MRSA isolates, predominantly livestock-associated MRSA ST398 was found, although 27·7% belonged to ST9 (spa type t1430). There is an increased risk of MRSA carriage in personnel working at broiler slaughterhouses, particularly those having contact with live animals.