This study has assessed the ability of five hand-washing agents, Hibisol, Hibiscrub, Betadine, 70% isopropyl alcohol and bar soap, to decontaminate the surfaces of Biogel D, Surgikos Microtouch and Ansell Gammex latex medical gloves after repeated inoculation with Staphylococcus aureus. The glove and hand-washing agents were compared in a Latin Square experiment, and a standardized handwashing regime followed. Each of the gloves used in the study was later tested electrically for micropuncture formation. On the basis of microbial recovery, no specific combination of glove and hand washing agent proved superior, the marker organism being isolated on a sporadic basis. In addition, 17% of the gloves showed evidence of micropunctures after five sequential inoculations and washings. These results suggest that, even under strictly controlled conditions, glove surfaces cannot be reliably and consistently cleaned of microbial contamination, nor can they be guaranteed to remain puncture-free. Ideally, therefore, a new pair of gloves should be worn for each patient. © 1990.