Duplex real-time PCR combined with melting curve analysis for rapid detection of Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss)
2021
Xu, Wenjie | Fu, Mingyang | Huang, Manhong | Cui, Xiaowen | Li, Yi | Cao, Min | Wang, Libin | Xiong, Xiaohui | Xiong, Xiong
Financial loss and health risk caused by the substitution with Salmo salar and Oncorhynchus mykiss have been widely reported around the world, highlighting the necessity to establish rapid and accurate methods for species identification. The aim of the present study was to develop a novel method for rapid identification of S. salar and O. mykiss based on duplex PCR assay (conventional and fluorescent). Specifically, after sequence alignment, specific primers for S. salar and O. mykiss based on the COⅠ (cytochrome oxidase I) and cyt b (cytochrome b) genes were designed and the specificity was verified against 23 common fish species. Clear and distinguishable bands of the expected sizes of 108 bp, and 207 bp, as well as the melting peaks around 81.6 ℃ and 84.7 ℃, were observed for S. salar, and O. mykiss, respectively. Moreover, the feasibility of melting curve analysis for detecting the adulteration of different amounts of O. mykiss with S.salar has also be confirmed. Therefore, the novel assay in the present study allows a fast and accurate identification of S. salar and O. mykiss in processed fish products.
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