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: <i>Argopecten irradians</i>, <i>Mimachlamys crassicostata</i>, <i>Mizuhopecten yessoensis</i>, <i>Scaeochlamys farreri</i>, <i>Atrina pectinata</i>, and <i>Swiftopecten swiftii</i>. 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.