Mackerel is marketed at prices according to the species type, Scomber japonicus and Scomber scombrus. Distinguishing these two species with the naked eye is difficult, and their differentiation becomes more difficult after they are processed by cooking, thereby leading to counterfeiting issues. Thus, in this study, we developed a method to differentiate S. japonicus from S. scombrus by detecting polymorphisms in mitochondrial 16 s rRNA gene by using fluorescence melting curve analysis and locked nucleic acid probes. Our method could distinguish S. japonicus from S. scombrus in a single experiment by using a single probe. The probes developed matched exactly with S. japonicus and had a melting temperature of 64 °C. However, the probes were mismatched with S. scombrus, resulting in a lower melting temperature of 46 °C. The high specificity of the locked nucleic acid probes resulted in this large difference in the melting temperatures.
Keywords: Fluorescence melting curve analysis; Locked nucleic acid; Scomber japonicus; Scomber scombrus; species identification.