title: Seafood Identification in Multispecies Products: Assessment of 16SrRNA, cytb, and COI Universal Primers' Efficiency as a Preliminary Analytical Step for Setting up Metabarcoding Next-Generation Sequencing Techniques.
abstract: Few studies applying NGS have been conducted in the food inspection field, particularly on multispecies seafood products. A preliminary study screening the performance and the potential application in NGS analysis of 14 "universal primers" amplifying 16SrRNA, cytb, and COI genes in fish and cephalopods was performed. Species used in surimi preparation were chosen as target. An in silico analysis was conducted to test primers' coverage capacity by assessing mismatches (number and position) with the target sequences. The 9 pairs showing the best coverage capacity were tested in PCR on DNA samples of 53 collected species to assess their amplification performance (amplification rate and amplicon concentration). The results confirm that primers designed for the 16SrRNA gene amplification are the most suitable for NGS analysis also for identification of multispecies seafood products. In particular, the primer pair of Chapela et al. (2002) is the best candidate.
authors.affiliation: FishLab, Department of Veterinary Sciences, University of Pisa , Via delle Piagge 2, 56124 Pisa, Italy.
authors.forename: Alice
authors.initials: A
authors.lastname: Giusti
authors.affiliation: FishLab, Department of Veterinary Sciences, University of Pisa , Via delle Piagge 2, 56124 Pisa, Italy.
authors.forename: Lara
authors.initials: L
authors.lastname: Tinacci
authors.affiliation: Instituto de Investigaciones Marinas (IIM-CSIC) , Eduardo Cabello 6, 36208 Vigo, Spain.
authors.forename: Carmen G
authors.initials: CG
authors.lastname: Sotelo
authors.affiliation: FishLab, Department of Veterinary Sciences, University of Pisa , Via delle Piagge 2, 56124 Pisa, Italy.
authors.forename: Martina
authors.initials: M
authors.lastname: Marchetti
authors.affiliation: FishLab, Department of Veterinary Sciences, University of Pisa , Via delle Piagge 2, 56124 Pisa, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Guidi
authors.affiliation: Tianjin Entry-Exit Inspection and Quarantine Bureau of the People's Republic of China , Jingmen Road 158, Free trade Zone, Tianjin Port, 300461 Tianjin, China.
authors.forename: Wenjie
authors.initials: W
authors.lastname: Zheng
authors.affiliation: FishLab, Department of Veterinary Sciences, University of Pisa , Via delle Piagge 2, 56124 Pisa, Italy.
authors.forename: Andrea
authors.identifier: http://orcid.org/0000-0002-9614-2032
authors.initials: A
authors.lastname: Armani
mesh_terms: D000818:Animals; D045303:Cytochromes b / Q000235:genetics*; D058893:DNA Barcoding, Taxonomic / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D049832:Decapodiformes / Q000145:classification / Q000235:genetics*; D003576:Electron Transport Complex IV / Q000235:genetics*; D029941:Fish Proteins / Q000235:genetics; D005399:Fishes / Q000145:classification / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D017747:Seafood / Q000032:analysis* / Q000145:classification
chemical_list: D017931:DNA Primers; D029941:Fish Proteins; D012336:RNA, Ribosomal, 16S; D045303:Cytochromes b; D003576:Electron Transport Complex IV
keywords: cephalopods; fish; metabarcoding; multispecies seafood products; next-generation sequencing; universal primers