Mushroom mites have recently caused severe damage to sawdust beds, mushroom mycelia, and fruiting bodies; therefore, they have reduced the production of the button mushroom, Agaricus bisporus, in greenhouses. There are currently no registered pesticides for mushroom mites. It is necessary to selectively control mushroom flies and mites without affecting the growth of the mushroom. We examined biological control of mushroom mites using predatory mite Stratiolaelaps scimitus in button mushroom cultivation. As a result, a three times treatment (1 treatment after water cleaning, 1 treatment after fungus inoculation, and 1 treatment before or after casing) was most effective at controlling mushroom mites, with 3.000 predatory mites (3 bottles) scattered evenly over 165–230 ㎡ every 1–2 m. Predatory mite Stratiolaelaps scimitus could control mushroom flies and mites at the same time and could be used at any time during cultivation.