Ireland had the highest prostate cancer incidence in Europe in 2006. In that year, the National Cancer Forum (NCF) recommended against prostate specific antigen (PSA) testing for population-based screening. To investigate (1) PSA services and (2) impact of the NCF recommendation. Questionnaires were dispatched to biochemistry laboratories nationwide in 2006 and 2007. All 55 laboratories responded in 2006; 33/36 (89%) responded in 2007. 36 laboratories measured total PSA (tPSA); 14 measured free PSA (fPSA). Laboratories with higher tPSA workload were more likely to measure fPSA (P = 0.024). A total of 15 laboratories used age-specific PSA ranges. In 2006, there were [382,000 tPSA and [48,000 fPSA tests costing an estimated euro 4,900,000. During 2006-2007 tPSA tests increased by 11%; fPSA tests decreased by 36%. There is considerable inter-laboratory variation in PSA testing practices. Because of the potential clinical consequences, standardisation should be considered. Testing practice was unaffected by the NCF recommendation.