Development of a Multiplex PCR Assay for Simultaneous Identification of Six Commercially Important Bivalves

Processed bivalve products typically lack diagnostic shell characters, making DNA-based assays essential for reliable species authentication in routine testing. We developed and validated a conventional, COX1-based multiplex PCR for the simultaneous identification of six commercially important food bivalves: Argopecten irradians, Mimachlamys crassicostata, Mizuhopecten yessoensis, Scaeochlamys farreri, Atrina pectinata, and Swiftopecten swiftii. The assay yielded robust, single-band amplification across an annealing window of 54–59 °C and maintained reproducible performance over template inputs from 0.1 to 50 ng per reaction. This work provides a practical marker set for simultaneous identification of six bivalve species in freeze foods or tissues where morphological traits are not preserved.