This invention discloses a melting curve primer-probe set and detection method for detecting harmful dinoflagellates, as well as its application. The melting curve primer-probe set includes a first primer, a medium probe, and / or a second primer and a
TaqMan probe. The detection method includes the following steps: S1, performing an amplification reaction; S2, if the
nucleic acid to be tested contains one of AP, MP, KM, AAN, or GC, both the upstream and downstream primers of the first primer are extended; S3, forming a double-stranded product suitable for melting curve detection; S4, if the
nucleic acid to be tested contains SA and an internal
reference gene, both the upstream and downstream primers of the second primer are extended; S5, generating an amplification curve; S6, analyzing the melting curve and amplification curve of the double-stranded product to determine the detection result of harmful dinoflagellates. The melting curve primer-probe set can be used for the detection of harmful
dinoflagellate content.