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We design a ratiometric fluorescent sensing platform for bleomycin (BLM) by using proximity-dependent DNA-templated silver nanoclusters (DNA-AgNCs) probe.This ratiometric sensing system is constructed with DNA-AgNCs as single fluorophore.The proposed strategy is based on the two following facts: (1) a covert DNA can approach and transform the DNA-AgNCs with green emission (G-DNA-AgNCs) into red emission through hybridization reaction.(2) The specific cleavage of the convert DNA by BLM in the presence of Fe(Ⅱ) inhibits the discoloration of G-DNA-AgNCs.Thus,benefiting from the specific recognition of BLM and unique properties ofG-DNA-AgNCs,a highly-sensitive ratiometric sensor for BLM has been successfully developed.The detection limit is as low as 30 pmol/L.This label-free fluorescence probe possesses advantages of convenient synthetic process and low cost.Moreover,this ratiometric method has been applied to the detection of BLM in human serum samples,illustrating a promising tool for analysis of BLM in cancer therapy.