Hospital linen is clearly recognized as a potential reservoir for microorganisms and could be a vector of disease transmission. The aim of this study was to isolate, count and identify fungi and bacteria from different kinds of clean and dirty linen in a hospital. Microbiological samples have been collected on clean bed linen (n = 200), dirty bed linen (n = 192) and staff uniforms (n = 192) by using contact plates. 55% of samples from clean bed linen were contaminated before contact with the patient, with a mean count of 3 cfu/25 cm² (range: 1-117 cfu) when contaminated. Virtually all samples from dirty bed linen carried microorganisms, with a mean count of 23 cfu/25 cm² (range 1-191 cfu). In addition, staff hospital uniforms showed the highest contamination rates in the study, with an average of 45 cfu/25 cm² (range: 1-218 cfu). Microbial species were mostly bacteria commonly found in the environment or on human skin, such as staphylococci or micrococci. Nevertheless, 57% of the identified species may be opportunistic pathogens for humans, representing a risk for people with a deficient or weakened immune system, especially in cases of superinfection. Since contamination of linen seems to occur after washing, actively antimicrobial textiles would represent a valuable measure to prevent textiles from being a vehicle for transfer of microorganisms.