Detection of DNA damage is significant for the evaluation of genotoxicity of new chemicals in the early stages of its development. An electrogenerated chemiluminescence (ECL) biosensor was fabricated to detect specific sequences of DNA by using CdTe@SiO₂ as nanoprobes for signal amplification. This DNA biosensor was constructed by self-assembly of an aminated capture DNA on the glass carbon electrode. DNA detection was realized by outputting a remarkable ECL signal of the CdTe@SiO₂ labeled probe DNA. When the target DNA was introduced into the system, it was complementary to the probe DNA at the one-half-segment and complementary to the capture DNA at the other half-segment, resulting in the formation of a stable duplex complex. As a result, the CdTe@SiO₂ labeled probe was proximate to the electrode surface and the ECL was observed. This DNA biosensor was proved to have a low detection limit (0.03 nM) and a wide dynamic range (from 0.1 nM to 2 μM). Most importantly, the sensing system could differentiate the single base mismatched DNA from the complementary DNA. It was successfully applied to study the damage to DNA caused by several genotoxicity chemicals, which was rapid, simple, reliable and sensitive compared to the classical biological methods.