A New Rapid Method for the Authentication of Common Octopus (Octopus vulgaris) in Seafood Products Using Recombinase Polymerase Amplification (RPA) and Lateral Flow Assay (LFA)

Abstract

The common octopus (Octopus vulgaris) is a highly valued cephalopod species which is marketed with different grades of processing, such as frozen, cooked or even canned, and is likely to be mislabeled. Some molecular methods have been developed for the authentication of these products, but they are either labor-intensive and/or require specialized equipment and personnel. This work describes a newly designed rapid, sensitive and easy-to-use method for the detection of Octopus vulgaris in food products, based on Recombinase Polymerase Amplification (RPA) and a detection using a Lateral Flow assay (LFA). After studying several gene markers, a system of primers and nfo-probe was designed in the COI (Cytochrome Oxidase I) region and was successfully tested in 32 reference samples (covering 14 species) and 32 commercial products, after optimization. The method was also validated in a ring trial with eight European laboratories and represents a useful tool for food authenticity control at all levels of the value chain.

Phylogeography of the veined squid, Loligo forbesii, in European waters

“…We found a total of 16 COI-haplotypes for L. forbesii, whereof three have been reported before [32][33][34][35][36][37][38][39][40] (H7 and H10, "East Atlantic" clade; H25, Azorean clade, see Fig. 3).…”

“…The sequences obtained were analysed with the software "CEQ8000 Genetic Analysis System" (Version 9.0.25, Beckman Coulter GmbH, Germany) and an alignment of 249 sequences was created ("BioEdit" version 7.2.5. 32 ) of which 28 were retrieved from GenBank [32][33][34][35][36][37][38][39][40] . The phylogenetic analyses (Maximum Likelihood, ML) was performed using "MEGA" version 6 41 with 1000 bootstraps.…”

Species authentication of canned mackerel: Challenges in molecular identification and potential drivers of mislabelling

“…2021) reporting 100% mislabelling in five canned Zamburiñas (Mimachlamys varia) analyzed. All the other cases, including herrings, anchovies, tuna and octopus, showed a lower mislabelling rate, ranging from 0% to 16.9% (Chang et al, 2021;Chin Chin et al, 2016;Giusti et al, 2019;Nagalakshmi et al, 2016;Sotelo et al, 2018;Tinacci et al, 2019;Velasco et al, 2021). This contrasting scenario could be generated by the limited number of studies focused on canned seafood, but also to the weak and often ambiguous European legislation, which allows the use of generic information of species of fish used.…”

Assessing the quality of octopus: From sea to table

“…In this regard, squid ( Dosidicus gigas ) is usually used for sale because it is a species of lower commercial value but morphologically very similar to those of the octopus (Pardo et al., 2018). This has forced the development of different methods that allow the authentication of common octopus ( O. vulgaris ), such as Forensically informative nucleotide sequencing and barcoding, PCR‐RFLP (restriction fragment length polymorphism), real time PCR, RAPD (random amplified polymorphic DNA), recombinase polymerase amplification, and lateral flow assay (Lee et al., 2022; Velasco et al., 2021).…”