A Fast and Simple DNA Mini-barcoding and RPA Assay Coupled with Lateral Flow Assay for Fresh and Canned Mackerel Authentication

Abstract

Nowadays, food authentication is more and more required given its relevance in terms of quality and safety. The seafood market is heavily affected by mislabelling and fraudulent substitutions/adulterations, especially for processed food products such as canned food items, due to the loss of morphological features. This study aims to develop new assays based on DNA to identify fresh mackerel ( Scomber spp.) and commercial products. A new primer pair was de novo designed on the 5S rRNA gene and non-transcribed spacer (NTS), identifying a DNA mini-barcoding region suitable for species identification of processed commercial products. Moreover, to offer a fast and low-cost analysis, a new assay based on recombinase polymerase amplification (RPA) was developed for the identification of fresh ‘Sgombro’ ( Scomber scombrus ) and ‘Lanzardo o Occhione’ ( Scomber japonicus and Scomber colias ), coupled with the lateral flow visualisation for the most expensive species ( Scomber scombrus ) identification. This innovative portable assay has great potential for supply chain traceability in the seafood market. Graphical Abstract Supplementary Information The online version contains supplementary material available at 10.1007/s12161-022-02429-6.

Convergent technologies to tackle challenges of modern food authentication

“…In this context, an effective simplification has recently occurred for DNA-based methods. Biomolecular methods have become increasingly simple and immediate [ 13 ] especially given to the use of target PCR analyses to detect specific contaminants and/or to certify the species origin. Simplified techniques such as the Loop-mediated isothermal amplification (LAMP), Recombinase Polymerase Amplification (RPA), and HDA Helicase-dependent Amplification) comply with these requirements, allowing for rapid food analysis (less than 2 h) even when equipped laboratory and instruments (e.g., thermocyclers) are not available.…”

Molecular Barcoding: A Tool to Guarantee Correct Seafood Labelling and Quality and Preserve the Conservation of Endangered Species

“…Almost 85% of the analysed products contained highly allergenic hake or grenadier species and only 15% contained less histaminergic species such as cod and haddock [ 148 ]. Frigerio et al [ 149 ] performed a new set of primers on the 5S rDNA and non-transcribed spacer ( NTS ) on 27 processed and unprocessed products collected at the Milan fish market and across different Italian supermarkets. In their research, a new DNA mini-barcoding region suitable for species identification was identified [ 149 ].…”

“…Frigerio et al [ 149 ] performed a new set of primers on the 5S rDNA and non-transcribed spacer ( NTS ) on 27 processed and unprocessed products collected at the Milan fish market and across different Italian supermarkets. In their research, a new DNA mini-barcoding region suitable for species identification was identified [ 149 ].…”

Establishment of rapid extraction and sensitive detection system of trace corn syrup DNA in honey

“…Recombinant enzyme polymerase amplification (RPA) has become a suitable technique for species identification due to its simplicity, short reaction time, and low reaction temperature ( Tian, Liu, Ye, Li, & Chao, 2017 ). RPA has been used to authenticate animal products, including meat ( Cao et al, 2018 , Ivanov et al, 2021 , James and Macdonald, 2015 , Kissenkoetter et al, 2020 ) and seafood ( Frigerio et al, 2023 , Xie et al, 2023 ). There are few reports of its use to detect honey adulteration.…”