High-Resolution Melting (HRM) Analysis for Rapid Molecular Identification of Sparidae Species in the Greek Fish Market

Evanthia Chatzoglou; Nefeli Tsaousi; Apostolos P Apostolidis; Athanasios Exadactylos; Raphael Sandaltzopoulos; Ioannis A Giantsis; Georgios A Gkafas; Emmanouil E Malandrakis; Joanne Sarantopoulou; Maria Tokamani; George Triantaphyllidis; Helen Miliou

doi:10.3390/genes14061255

High-Resolution Melting (HRM) Analysis for Rapid Molecular Identification of Sparidae Species in the Greek Fish Market

Genes (Basel). 2023 Jun 12;14(6):1255. doi: 10.3390/genes14061255.

Affiliations

¹ Laboratory of Applied Hydrobiology, Department of Animal Science, School of Animal Biosciences, Agricultural University of Athens, 11855 Athens, Greece.
² Laboratory of Fish & Fisheries, Department of Animal Production, School of Agriculture, Faculty of Agriculture, Forestry and Natural Environment, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
³ Hydrobiology-Ichthyology Lab, Department of Ichthyology and Aquatic Environment, School of Agricultural Sciences, University of Thessaly, 38446 Volos, Greece.
⁴ Department of Molecular Biology and Genetics, Democritus University of Thrace, 68100 Alexandroupolis, Greece.
⁵ Department of Animal Science, Faculty of Agricultural Sciences, University of Western Macedonia, 53100 Florina, Greece.

Abstract

The red porgy (Pagrus pagrus) and the common dentex (Dentex dentex) are Sparidae species of high commercial value, traded in the Greek market. In some cases, fish species identification from Greek fisheries is difficult for the consumer due to the strong morphological similarities with their imported counterparts or closely related species such as Pagrus major, Pagrus caeroleustictus, Dentex gibbosus and Pagellus erythrinus, especially when specimens are frozen, filleted or cooked. Techniques based on DNA sequencing, such as COI barcoding, accurately identify species substitution incidents; however, they are time consuming and expensive. In this study, regions of mtDNA were analyzed with RFLPs, multiplex PCR and HRM in order to develop a rapid method for species identification within the Sparidae family. HRM analysis of a 113 bp region of cytb and/or a 156 bp region of 16s could discriminate raw or cooked samples of P. pagrus and D. dentex from the aforementioned closely related species and P. pagrus specimens sampled in the Mediterranean Sea when compared to those fished in the eastern Atlantic. HRM analysis exhibited high accuracy and repeatability, revealing incidents of mislabeling. Multiple samples can be analyzed within three hours, rendering this method a useful tool in fish fraud monitoring.

Keywords: 16s; COI; Dentex; Pagrus; authentication; barcoding; cytb; fish mislabeling; fraud; mtDNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
DNA, Mitochondrial / genetics
Greece
Mitochondria / genetics
Perciformes* / genetics
Polymerase Chain Reaction

Substances

DNA, Mitochondrial

Grants and funding

This work was supported by the Greek Operational Programme Maritime & Fisheries 2014–2020, which is co-financed by the EU European Maritime and Fisheries Fund and the Hellenic Republic Ministry of Rural Development and Food, under the title: MOLECULAR IDENTIFICATION OF FISH AUTHENTICITY AND ADULTΕRATION IN GREEK MARKET—FISH MID (MIS 5033599).