A quantitative polymerase chain reaction approach for distinguishing the geographical origin of Pagrus major from Korea and Japan

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Abstract
Illegal distribution and mislabeling of imported fishery products pose challenges to country-of-origin labeling regulations in Korea. To address this issue, a quantitative PCR (qPCR)-based method was developed to distinguish Pagrus major from Korea and Japan. Using genotyping-by-sequencing (GBS), two single nucleotide polymorphism (SNP) markers were identified, and allele-specific primers were designed. Gaussian mixture modeling established Ct thresholds, achieving accuracy of 81.67% and 77.78% for SNP001 and SNP008, respectively. The amplification efficiency and limit of detection (LOD) were assessed using tenfold serial dilutions (10-0.001 ng/μL). Standard curves for AA and TT homotypes showed high linearity (R2 > 0.994) with amplification efficiencies of 103.65% and 97.63%, respectively. This qPCR-based method provides a reliable approach for origin verification of P. major, aiding regulatory enforcement and ensuring seafood authenticity. The online version contains supplementary material available at 10.1007/s10068-025-01877-0.
Funding Information
  • Korea Institute of Marine Science and Technology promotion (RS-2022-KS221671)

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