The DNA templated silver nanoclusters were prepared by reducing silver nitrate with sodium borohydride as reducing agent to detect HBV sequence. The factors influencing the fluorescence of silver nanoclusters were explored. Under the optimized experimental conditions, the HBV sequence was analyzed with 466 nm as excitation wavelength. The probe had a good selectivity for HBV sequence. There is a good linear relationship between 0.05 μM and 1 μM and limit of detection is 0.016 μM.