Exemplary embodiments relate to a microfluidic structure including: a plurality of sample chambers; a reaction chamber in which at least two types of materials, which respectively specifically react with at least two types of target materials, are immobilized; a detection chamber connected to the reaction chamber; a path connecting the chambers; and a valve for opening and closing the path, and a microfluidic device including the microfluidic structure. Since at least two types of materials specifically binding to target materials are immobilized in a reaction chamber of the microfluidic structure, space may be efficiently used and the target materials may be assayed in a one-step test. An internal space of the microfluidic device using the microfluidic structure, the amount of samples, and costs for manufacturing the microfluidic device may be reduced, and internal quality control may be efficiently performed using the microfluidic structure as a control for the operations.