Significant progress on upconversion-nanoparticle (UCNP)-based probes is witnessed in recent years. Compared with traditional fluorescent probes (e.g., organic dyes, metal complexes, or inorganic quantum dots), UCNPs have many advantages such as non-autofluorescence, high chemical stability, large light-penetration depth, long lifetime, and less damage to samples. This article focuses on recent achievements in the usage of lanthanide-doped UCNPs as efficient probes for biodetection since 2014. The mechanisms of upconversion as well as the luminescence resonance energy transfer process is introduced first, followed by a detailed summary on the recent researches of UCNP-based biodetections including the detection of inorganic ions, gas molecules, reactive oxygen species, and thiols and hydrogen sulfide.