A water-soluble polymeric pyrene-based polythioacetal (PTA-Py) with thioacetal units in the main chain is simply synthesized by direct polycondensation of 3, 6-dioxa-1, 8-octanedithiol, 1-pyrene formaldehyde, and mPEG2k-SH. The probe PTA-Py shows a good fluorescence response to Hg2+ ions due to the Hg2+-promoted deprotection reaction of thioacetal groups to regenerate the original 1-pyrene formaldehyde compound. After adding Hg2+ to the PTA-Py solution, the fluorescence intensity (FI) gradually increases with increasing concentrations of Hg2+. Compared with other metal ions, the probe exhibits high sensitivity, good selectivity, and rapid response to Hg2+. The low detection limits are 12.3nm in ethanol-PBS buffer and 13.3nm in water, respectively. The results imply that the simply synthesized water-soluble polymeric probehad potential applications in the rapid detection of Hg2+ ions in aqueous solutions. Moreover, the polymeric PTA-Py shows high sensitivity for CH3Hg+ with detection limits of 26.5nm in ethanol/PBS buffer. In addition, PTA-Py can efficiently detect Hg2+ ions in HeLa cells. The results demonstrate that a valuable method is developed for biocompatible polymeric sensors for Hg2+ ions in biological and environmental samples.