A microorganism culture sheet is provided which can limit the use of a costly cultivation material to a necessary portion, which has simple production at the time of manufacture, and can be simply and stably used by a user. The microorganism culture sheet of the present invention is a microorganism culture sheet comprising a base sheet, a culture layer formed on top of the base sheet, and a cover sheet that covers the culture layer, and the culture layer is pattern-formed with an application liquid comprising a polyvinylpyrrolidone and at least on selected from the group consisting of a gelling agent, a nutrient component, a color indicator, a selective agent and a substrate.