We have developed a green approach to prepare DNA-templated silver nanoparticles (Ag-NPs) from the direct reaction between Ag and ascorbic acid in the presence of DNA and sodium hydroxide. The Ag-NPs showed strong resonance light scattering (RLS) intensity property. Then, the interaction between hydrogen sulfide (HS) and Ag-NPs was studied by measuring their RLS spectra. The results showed that there is a strong interaction between Ag-NPs and HS, which resulted in a decrease in the size of Ag-NPs and a decrease in the RLS intensity of the Ag-NPs solution at the wavelength of 467 nm. The results demonstrated that the RLS technique offers a sensitive and simple tool for investigating the interaction between Ag-NPs and HS, which can be applied to detect HS with high sensitivity and selectivity without complex readout equipment. The linear range for HS determination was found to be the range from 5.0 × 10 to 1.0 × 10 mol L, and the detection limit (3σ/k) was 2.8 × 10 mol L. Moreover, the proposed method was applied for the determination of HS in natural water samples with satisfactory results. Graphical abstract The application of Ag-NPs in HS detection.