Development of a SYBR Green-based quantitative PCR assay for the detection of Undaria pinnatifida and Endarachne binghamiae using chloroplast markers in processed seaweed products

초록

The global seaweed industry has expanded rapidly owing to increasing demand for sustainable food resources and bioactive compounds. Undaria pinnatifida, a widely cultivated brown seaweed in East Asia, possesses nutritional and medicinal properties; however, it is often misidentified as the morphologically similar non-commercial brown alga Endarachne binghamiae, leading to accidental admixture during harvesting and processing. Herein, we developed a SYBR Green-based quantitative PCR (qPCR) assay targeting species-specific single-nucleotide polymorphisms and insertions/deletions to accurately distinguish the two species. Six primer sets were designed and validated, yielding correlation coefficients over 0.99, amplification slopes of - 3.23 to - 3.43, and efficiencies of 95.41%-103.98%. The primers exhibited high specificity without false-positive reactions when tested against two target and 13 non-target seaweed species. The assay was successfully applied to 15 commercial U. pinnatifida products. This qPCR assay is a rapid, sensitive, and cost-effective tool for authenticating similar seaweed species, ensuring food safety and quality.

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