The Xiamen Symposium on Marine Environmental Sciences

2027 Xiamen Symposium on Marine Environmental Sciences

January 12-15, 2027

Development of a rapid RPA-CRISPR/Cas12a-LFD assay for the detection of toxin-producing harmful algal bloom species
ID:35 Oral Presentation

2027-01-13 15:35 (China Standard Time)

Session:Session 2 - Harmful Algal Blooms in the Asia Pacific: Recent Advances in Taxonomy, Biodiversity, Ecophysiology, Rapid Detection, and Early Warning to Mitigate Impacts of Coastal Ecosystem Health and Seafood Safety

Abstract
Harmful algal blooms (HABs) pose severe threats to marine ecosystems, aquaculture productivity, and public health worldwide, yet rapid on-site detection for toxigenic species remains technically challenging. Here, we developed a diagnostic assay integrating recombinase polymerase amplification (RPA) with CRISPR/Cas12a and lateral flow dipstick (LFD) for the visual detection of Pseudo-nitzschia multistrata, a  toxigenic diatom species responsible for the production of neurotoxin domoic acid (DA). The assay targets the dabA gene, a functionally critical component of the DA biosynthesis gene cluster, whose detection serves as a molecular proxy for the toxigenic potential of the species. Following a simple alkaline lysis step, the released DNA was subjected to RPA for pre-amplification of the dabA gene at 39℃ for 35 minutes. The RPA product then served as the substrate for CRISPR/Cas12a reactions, in which the Cas12a protein, guide RNA, and a fluorescently labeled ssDNA probe were incubated at 37℃ for 30 min. Cas12a specifically recognized the target amplicon and activated its trans-cleavage activity, resulting in the cleavage of the probe. The RPA-CRISPR products were subsequently applied to LFD strips, with results visually interpreted within 5 min. The entire workflow, from cell lysis to result readout, can be accomplished within 1.5 hours without requiring thermal cycling equipment. This method offers a rapid, equipment-independent, and user-friendly approach for the detection of toxigenic HAB species, providing a technical foundation for early warning and routine monitoring of HAB events in coastal ecosystems.
 
Speaker
Chao Gao Institute of Oceanology, Chinese Academy of Sciences

Author
Chao Gao Institute of Oceanology, Chinese Academy of Sciences

    Important Dates

    • Jan 30

      2026

      Session / Event proposals open

    • Mar 31

      2026

      Session / Event proposals close
      (Extended to April 7)

    • Apr 30

      2026

      Session acceptance notifications

    • Apr 30

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    • Jun 30

      2026

      Abstract submissions close
      (Extended to July 21)

    • Aug 31

      2026

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    • Sep 15

      2026

      Scientific program released

    • Oct 15

      2026

      Early bird registration closes

    • Jan 12–15

      2027

      XMAS 2027