The stable and cadmium-free DNA functionalized ZnS:Mn2+ QDs were successfully synthesized through a facile one-step route. We constructed a novel FRET system based on one-step synthesized DNA-ZnS:Mn2+ QDs (donor) and WS2 nanosheets (acceptor). The FRET-based strategy was applied for the detection of streptavidin and folate receptor by combining TPSMLD and Exo III.