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).

The recent surge in algal blooms, known as Harmful Algal Blooms (HABs), poses significant ecological and economic challenges globally. Consequently, there is a growing emphasis on regular coastal surveillance to promptly identify HAB species and 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. However, 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 18 S rRNA gene copies per cell, ranging from 1 to 100 cells, by implementing capillary pipetting isolation procedure specifically for Prorocentrum triestinum species. A strong positive correlation of (r2 = 0.9923) was observed during the quantification of 18 S rRNA gene copies in 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. This research effectively demostrated the development of primers for identifying the Prorocentrum genus and the precise quantification of 18 S rRNA copies within Prorocentrum triestinum cells using ddPCR technology.