The extraordinary properties of layered graphene and its successful applications in electronics, sensors, and energy devices have inspired and renewed interest in other two-dimensional (2D) layered materials. Particularly, a semiconducting analogue of graphene, molybdenum disulfide (MoS2), has attracted huge attention in the last few years. With efforts in exfoliation and synthetic techniques, atomically thin films of MoS2 (single- and few-layer) have been recently prepared and characterized. 2D MoS2 nanosheets have properties that are distinct and complementary to those of graphene, making it more appealing for various applications. Unlike graphene with an indirect bandgap, the direct bandgap of single-layer MoS2 results in better semiconductor behavior as well as photoluminescence, suggesting its great suitability for electronic and optoelectronic applications. Compared to their applications in energy storage and optoelectronic devices, the use of MoS2 nanosheets as a sensing platform, especially for biosensing, is still largely unexplored. Here, we present a review of the preparation of 2D atomically thin MoS2 nanosheets, with an emphasis on their use in various sensing applications.