A Quantitative Polymerase Chain Reaction Method for Distinguishing Anadara kagoshimensis from Korea and China

The illegal distribution and misrepresentation of imported fishery products present significant obstacles to effective country-of-origin labeling enforcement in Korea. In this study, molecular markers based on single-nucleotide polymorphisms (SNP) were developed and validated to distinguish between Korean and Chinese populations of Anadara kagoshimensis (Tokunaga, 1906). A candidate SNP marker was identified by genotyping-by-sequencing (GBS) and an allele-specific primer was designed. The cycle threshold was determined using Gaussian mixture modeling, resulting in an origin identification accuracy of approximately 93%. Quantitative polymerase chain reaction (qPCR) assay results were consistent across varying quantities of A. kagoshimensis genomic DNA. This approach, which combined GBS- and qPCR-based SNP marker validation, provides an effective and robust method for determining the country of origin of A. kagoshimensis.