Development of species-specific PCR for the identification of three grouper fish species (Epinephelus septemfasciatus, E. bruneus and E. akaara)

The aim of the study was to develop rapid, precise and species-specific method of the identification of three grouper species (Red-spotted grouper, Epinephelus akaara; Longtooth grouper, E. bruneus; Seven-band grouper, E. septemfasciatus), by using polymerase chain reaction (PCR) mediated molecular method. A common forward primer and three different species-specific primers were designed for all the three species from the fragment of mitochondrial cytochrome c oxidase I (COI) gene. The species-specific amplification patterns showed three different bands of basepair length of had 344 bp (Red-spotted grouper), 262 bp (Longtooth grouper), and 130 bp (Seven-band grouper) which were available in the DNA samples isolated from each verified species, unknown collections, and even in mixed processed samples. Thus PCR assay provides reliable, simple, easy and low cost method for species identification of these three species of groupers.