Combining a COI Mini-Barcode with Next-Generation Sequencing for Animal Origin Ingredients Identification in Processed Meat Product

Abstract

For revealing animal species in complex or adulterated processed meat product, we presented a method combining a novel cytochrome oxidase I (COI) mini-barcode with next-generation sequencing (NGS), which identifies various animal species (swine, bovine, Caprinae, and some of fish, shrimp, and poultry) accurately and efficiently in processed meat products. We designed a universal primer based on 140 sequences from 51 edible animal species. A mixture of 12 species raw meat samples were identified with the clone sequencing and also with a mini-barcode- (136 bp) combined NGS method, respectively. The mini-barcode of these 12 species was 100% identical to the target species sequence by Sanger sequencing. Compared to the clone sequencing method, the NGS method is superior in accuracy, sensitivity, and detection efficiency. Various edible animal species were identified in the species level both in the mixed samples and the 7 heavily processed food products. Moreover, some unlabeled species and dubious contamination were detected as well.

Research on quantitative detection technology of raccoon‐derived ingredient adulteration in sausage products

“…Pan et al. ( 2020 ) used DNA barcode technology combined with NGS to accurately identify the problem of various types of meat adulteration and even showed good detection performance in samples mixed with multiple types of meat and reprocessed samples. A real‐time loop‐mediated isothermal amplification method for detecting pig genes in meat products has been established by Cai et al., which is the first attempt to use real‐time LAMP to detect pig‐derived components in commercial products (Cai et al., 2020 ).…”

Is the metabarcoding ripe enough to be applied to the authentication of foodstuff of animal origin? A systematic review

“…The taxonomic coverage was evaluated for 19 out of the 24 primer pairs (79.2%), as the other 6 were patented (P11, P19), or the tested TG/TS were not included in a food category of interest (P16), or the original study in which the primers were designed was not available (P12, P22) (Table 3). In terms of tcTG, the highest taxonomic coverage was observed for P9 (designed by Sarri et al, 2014) and P20 (Kocher et al, 1989), with seven and six TG, respectively, followed by P6 (Horreo et al, 2013) with five TG, and P14 (forward by Folmer et al, 1994 and reverse by Pan et al, 2020) and P23 (Kitano et al, 2007), with four TG each (Table 3). Twelve out of the 19 primer pairs (63.2%) only covered 1 TG.…”

“…The number of replicates should be established based on the type of investigated matrix. In this case, it was especially seafood, ranging from canned tuna (Kappel et al, 2017;Klapper et al, 2023), surimi (Noh et al, 2021), mussel-based products (Gense et al, 2021) and commercial sea cucumber (Xing et al, 2021), and only in one case, heavily processed meat products (Pan et al, 2020). Given the higher number of commercial seafood species with respect to meat, the use of replicates is especially useful.…”

Rapid analysis technologies with chemometrics for food authenticity field: A review

“…NGS and DNA barcoding applications for food authentication have gradually increased, with current research focusing on animal-derived foods. The NGS and DNA barcoding combination clearly distinguished five kinds of mammals, two types of poultry, and 12 kinds of animal sources for meat products ( Dobrovolny et al, 2022 ; Pan et al, 2020 ). This approach can also distinguish seafood species, such as salmon and edible seaweed breeds.…”