title: Species identification of the Northern shrimp (Pandalus borealis) by polymerase chain reaction-restriction fragment length polymorphism and proteomic analysis.
abstract: Genomic and proteomic techniques for species identification of meat and seafood products are being widely used. In this study, a genomic approach was used to differentiate Pandalus borealis (the Northern shrimp), which belongs to the superfamily Pandaloidea, from 30 crustaceans consisting of 19 commercially relevant prawns/shrimps species that belong to the superfamily Penaeoidea, which include the families Penaeidae and Solenoceridae, and 11 other crustacean species, including prawns, shrimps, lobsters, and crabs. For this purpose, a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was designed based on the amplification of the 16S rRNA/tRNA(Val)/12S rRNA mitochondrial regions using the primers 16S-CruF and 16S-CruR. The 966-bp PCR products were produced and cleaved with the restriction enzymes AluI, TaqI, and HinfI, which provided species-specific restriction patterns. In addition, a proteomic approach, based on matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and electrospray ionization-ion trap (ESI-IT) mass spectrometry, was used to identify and characterize new P. borealis-specific peptides that could be useful as potential markers of this species in protein-based detection methods. To our knowledge, this is the first time a molecular method has been successfully applied to identify a wide range of prawn and shrimp species, including P. borealis, for either whole individuals or processed products. However, validation of the methods proposed here is required by applying them to a larger sample of individuals from different populations and geographic origins in order to avoid mainly false-negative results.
authors.affiliation: Department of Analytical Chemistry, Nutrition, and Food Science, School of Veterinary Sciences, University of Santiago de Compostela, E-27002 Lugo, Spain.
authors.forename: Ananias
authors.initials: A
authors.lastname: Pascoal
authors.forename: Ignacio
authors.initials: I
authors.lastname: Ortea
authors.forename: José M
authors.initials: JM
authors.lastname: Gallardo
authors.forename: Benito
authors.initials: B
authors.lastname: Cañas
authors.forename: Jorge
authors.initials: J
authors.lastname: Barros-Velázquez
authors.forename: Pilar
authors.initials: P
authors.lastname: Calo-Mata
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D008297:Male; D009124:Muscle Proteins / Q000302:isolation & purification; D033542:Pandalidae / Q000737:chemistry / Q000145:classification* / Q000235:genetics*; D010449:Peptide Mapping; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D040901:Proteomics; D017422:Sequence Analysis, DNA; D012758:Shellfish / Q000032:analysis / Q000145:classification; D013045:Species Specificity; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D017931:DNA Primers; D004272:DNA, Mitochondrial; D009124:Muscle Proteins