Development and Validation of a Duplex PCR for the Simultaneous Detection of Atlantic Salmon (Salmo salar) and Rainbow Trout (Oncorhynchus mykiss) in Processed Fish Products
Article
Figures
Metrics
Preview PDF
Reference
Related
Cited by
Materials
Abstract:
Anovel method for rapid identification of Salmo salar and Oncorhynchus mykiss based on duplex PCR assay (conventional and fluorescent assays) was developed. Specifically, after sequence alignment, specific primers for S. salar and O. mykiss based on the COⅠ and Cyt b genes were designed and verified. The duplex PCR was optimized, and finally established. Only S. salar and O. mykiss amplified specific bands of 108 bp and 207 bp, while the other 23 non-target species were not successfully amplified. The optimal amount of primers for S. salar and O. mykiss were 0.1 μmol/L and 0.4 μmol/L, and the melting temperature was 81.5 ℃ and 85 ℃, respectively. Among the 29 commercial products, six products were identified as S. salar and three were identified as O. mykiss. Finally, theremaining 17 commercial samples were all tested as negative in the duplex PCR assay, and the results were consistent with DNA barcoding method. The results showed that the duplex PCR method for S. salar and O. mykiss was specific and feasible. This study provided reliable technical method and theoretical basis for the adulteration identification of S. salar and O. mykiss species. Key words:
