Quick method for grouper species identification using real-time PCR

A convenient and quick method based on real-time polymerase chain reaction (PCR) was established to identify 10 grouper species in China. According to the sequence of mitochondrial cytochrome oxidase subunit I (COI) gene, primers and probe were designed. PCR amplifications of 129 bp of COI gene sequences from 10 grouper species of four genera (Epinephelus, Promicrops, Plectropomus and Cromileptes) were compared with that of 30 other fish species. PCR amplifications were only observed in 10 grouper species. The result suggested that real-time PCR can be a powerful tool for grouper species diagnoses. ► Real-time PCR primer pairs and a probe were designed according to the sequence of COI gene. ► COI amplification was only observed in 10 groupers but not the other 30 fish species. ► The detection limit is 0.01%–1% for 10 grouper species respectively. ► In 41 grouper fillet and surimi samples from markets and restaurants, the results were in agreement with declarations.