[Problem] To provide a target substance detection chip, a target substance detection device, and a target substance detection method, that can be manufactured easily in a small size at low costs with reduction of the number of parts involved in the detection chip constituted by an optical prism and a detection plate used for a SPR sensor and an optical waveguide mode sensor, that can detect a target substance quickly with high sensitivity, and in which an analyte liquid is easily delivered.;[Solution] A target substance detection chip of the present invention includes: a plate-like transparent base portion which allows light to pass therethrough; and a flow path which is formed in one surface of the transparent base portion as a groove and through which an analyte liquid verifying a presence of a target substance is delivered in a length direction of the groove, wherein the flow path is formed such that at least an electric field enhancement layer is disposed on an inner surface of a groove portion formed to at least partly have inclined surfaces appearing in cross section to be inclined at a gradient to the surface of the transparent base portion, and wherein a part or entirety of an uppermost surface of the groove which contacts the analyte liquid serves as a detection surface for the target substance.
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