Biology, Vol. 15, Pages 219: Multiplex qPCR Assay for Simultaneous Detection of Three PST-Producing Dinoflagellates in the East China Sea off Southern Korea

Biology doi: 10.3390/biology15030219

Authors:
Jung Soo Heo
Biet Thanh Tran
Keun-Yong Kim
Sunju Kim
Seok Hyun Youn
Tae Gyu Park

Paralytic shellfish toxins (PSTs) are produced by several toxic species of the dinoflagellate genera Alexandrium and Gymnodinium, and they pose significant threats to marine ecosystems and public health. Rapid and accurate detection of harmful algal blooms (HABs) is essential for effective management. In this study, we developed a multiplex quantitative real-time PCR (qPCR) assay targeting the 28S ribosomal DNA region to simultaneously detect three PST-producing dinoflagellates, Alexandrium catenella, A. pacificum, and Gymnodinium catenatum, in the East China Sea off southern Korea. Species-specific primers and hydrolysis probes labeled with distinct fluorophores were validated for simultaneous detection. The standard curves showed strong linearity (R2 > 0.99) and high amplification efficiencies (95.268–99.325%). No cross-reactivity was observed among the 20 non-target microalgal species. Field application of the assay using environmental DNA (eDNA) samples collected during spring successfully detected A. catenella and A. pacificum, whereas G. catenatum was not detected during the survey period. This multiplex qPCR assay provides a rapid and reliable molecular tool for early detection and spatial monitoring of potentially PST-producing dinoflagellates, supporting sustainable HAB management in East Asian coastal ecosystems.

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