참돔과 홍민어 판별을 위한 multiplex PCR 검사법의 개발과 검증

Abstract

Nowdays, seafood consumption has increased and also defective food case also have increased in seafood. There are case that red drum(Sciaenops ocellatus) was sold instead of red seabream(Pagrus major). This study attempted to develop and validate species-specific primers for P. major and S. ocellatus to prevent the sale of S. ocellatus as P. major. Primers for P. major was designed 12s rRNA and S. ocellatus was designed 16s rRNA. PCR reaction was initial denaturation at 95°C for 5 min, followed by 30 cycles of denaturation at 95°C for 20 s, annealing at 56°C for 20 s and extension at 72°C for 30 s, with a final extension at 72°C for 5 min. Multiplex PCR showed a 468 bp amplicon for P. major and a 181 bp amplicon for S. ocellatus, respectively. The sensitivity of P. major and S. ocellatus was present at 1 ng each. The developed primer was validated with 19 P. major food samples through internet sale. Using this monitoring method, fake seafood did not appear due to the agreement between the experimental results and the species. Hence, the developed multiplex PCR method were reliable to authenticate the P. major and S. ocellatus.