This application provides universal labeled primers for detection and amplification of nucleic acid molecules. These universal primers can be attached to the 5′-end of a target sequence-specific primer. In particular examples, the universal primer includes a labeled nucleotide flanked on both sides a nucleotide whose complement nucleotides changes a detectable signal from the label when the universal primer hybridizes with its complementary nucleic acid molecule. Also disclosed are methods of using the universal primer in nucleic acid amplification, such as real-time PCR.