Universal primer design for crustacean and bivalve-mollusc authenticity based on cytochrome-b gene

Fish and seafood authenticity is important to support traceability practices and protect the public from economic fraud and adulteration. Molecular-based techniques of PCR are known as the most common methods for identifying seafood species. Nevertheless, these techniques rely on the appropriate primer set designed to amplify specific DNA fragments on targeted species. For efficiency application on a wide range of species, a universal primer set is more valuable than a specific primer. The present study developed universal primers, especially for identifying crustaceans and molluscs based on the cytochrome b mitochondrial DNA (Cyt b). The initial primer pair of CytBL1/CytBH originally designed for fish species was applicable to amplify the Cyt b gene on most selected fish samples, but not for crustacean and mollusc samples. Based on annealing profile, sequence evaluation (92-100% similarity), and RT-PCR analysis, the universal primer couple of CytBL1C/CytBHW designed in the present study potentially applied to identify crustacean and mollusc samples, especially shrimp and bivalve-mollusc.