Standardization of a Method for the Detection of the Genus Prorocentrum, a Dinoflagellate Present in Harmful Algae Blooms (Habs) in San Jorge Antofagasta Bay by Digital Droplet Pcr (ddPCR)

Abstract The recent surge in algal blooms, known as Harmful Algal Blooms (HABs), is causing significant ecological and economic challenges globally. There is a growing emphasis on regular coastal surveillance to promptly identify HAB species in order to mitigate or minimize their adverse effects. The integration of molecular techniques alongside traditional microscopy-based methods has emerged as a promising approach for coastal monitoring. Nevertheless, the availability of such data is currently limited to a select group of species. In response to these obstacles, a novel protocol has been devised for the differentiation of dinoflagellates belonging to the genus Prorocentrum using droplet digital PCR (ddPCR) in both laboratory (Ls) and environmental (Es) samples. This study employed ddPCR technology to quantify the 18s rRNA gene copies per cell within a range of 1 to 100 cells by implementing an isolation procedure involving capillary pipetting specifically for the Prorocentrum triestinum species. A strong positive correlation of r2 = 0.9923 was observed in the quantification of the copies for P . triestinum cells. The copies per cell of P . triestinum determined through ddPCR for both Ls and Es samples were found to be approximately 2000 ± 176 and 1862 ± 136, respectively. The research effectively showcased the development of primers for identifying the Prorocentrum genus and the precise quantification of 18s rRNA copies within Prorocentrum triestinum cells using ddPCR technology.