Layered transition metal dichalcogenides (TMDs) are important members in the family of two-dimensional (2D) materials. The large surface-to-volume ratio, combined with the fascinating tunable electronic and optical properties, low toxicity, unique van der Waals layered structure, and engineerable surface structure, renders 2D TMDs highly valuable for next-generation biosensing applications. Herein, the recent progress in the development of 2D TMDs-based biosensors is comprehensively reviewed, with special focus on the implementation of the structural, electronic and optical properties of 2D TMDs in the realization of high-performance biosensors with different configurations for a wide spectrum of bioanalytes and bio-species. In addition, the comparison on biosensing performances with graphene as the currently most studied 2D candidate is critically discussed. Finally, future perspectives are provided along the development progress of 2D TMDs-based biosensors which are currently undergoing an intense study. This work will lead researchers to explore more novel sensing candidates within the category of TMDs with exotic chemical composition, structure, morphologies, dimensionalities, and properties. Display omitted •Structure-property relationship in transition metal dichalcogenides (TMDs).•The merits and demerits of two-dimensional TMDs when compared with graphene.•Solid-, liquid-, and vapor-phase-based synthesis of TMDs.•Fabrication of optical and electrical sensing platform based on TMDs.•Future perspectives and challenges of TMDs-based biosensors.