In this study, selected detrimental and essential elements were determined in fruiting bodies of Auricularia auricula-judae, Tylopilus felleus, and Pleurotus ostreatus medicinal mushrooms. A. auricula-judae and T. felleus were collected from sampling sites situated in South Bohemia, the Czech Republic, while P. ostreatus samples were obtained after cultivation under defined conditions on a straw substrate block. The elements were determined in underlying soils for T. felleus and bioconcentration factors were calculated. The analysis revealed that T. felleus accumulated Rb, Cd, Cu, Zn, Se, and Mg, with bioconcentration factors of 65.7, 2.47, 2.32, 1.93, 1.67, and 1.27, respectively. The analysis also revealed high contents (expressed as medians) of Ca (1600 mg/kg) and Mg (2040 mg/kg) in A. auricula-judae. On the other hand, the contents of detrimental elements such as Pb (0.10 mg/kg), Cd (0.33 mg/kg), and As (0.10 mg/kg) were lower. In cultivated P. ostreatus, lower element contents for Pb (< 0.05 mg/kg), Al (3.92 mg/kg), Ca (53.6 mg/kg), Mn (3.22 mg/kg), Ni (0.11 mg/kg), and Sr (0.93 mg/kg) were determined compared to both wild-growing A. auricula-judae and T. felleus. No negative effect on human health is expected for the mushroom species studied due to the accumulation of detrimental microelements.