In this work, a novel signal amplification biosensor was utilized to detect Cd based on asymmetric PCR. In the presence of Cd, it can bind with Cd-aptamer C1 which caused the complementary strand C2 to be released from double-stranded DNA C1-C2. Because the single-stranded C1 cannot be hydrolyzed by Exo III, it can be used as a template to take part in asymmetric PCR reaction.