Development of a multiplex PCR assay for the simultaneous detection of big blue octopus (Octopus cyanea), giant Pacific octopus (Enteroctopus dofleini), and common octopus (Octopus vulgaris).

Abstract

Since octopuses are similar in appearance and can be processed into various forms, seafood fraud has been reported. In this study, we developed the PCR assay to simultaneously detect three octopuses (big blue octopus, giant Pacific octopus, and common octopus). Specific primer sets were designed based on COI gene. We observed that the specific PCR amplicon sizes were 84bp for big blue octopus, 117bp for giant Pacific octopus, and 166bp for common octopus, respectively. This assay was then used to test for specificity and did not show cross-reactivity with 15 cephalopods families. The limit of detection of the multiplex PCR assay was 0.1pg. Subsequently, 30 commercial food products were then monitored to evaluate the applicability of this assay. All products were specifically amplified, and three octopus species of interest were distinguished. Therefore, this assay can be used as an octopus authentication tool in the seafood industry.