Differentiation of five tuna species by a multiplex primer-extension assay.
A novel methodology based on analysis of mtDNA-cytb diagnostic sites was performed to discriminate four closely related species of Thunnus (Thunnus alalunga, Thunnus albacares, Thunnus obesus and Thunnus thynnus) and one species of Euthynnus (Katsuwonus pelamis) genus in raw and canned tuna. The primers used in the preliminary PCR designed in well conserved region upstream and downstream of the diagnosis sites successfully amplified a 132bp region from the cytb gene of all the species taken into consideration. The sites of diagnosis have been interrogate simultaneously using a multiplex primer-extension assay (PER) and the results were confirmed by fragment sequencing. The applicability of the multiplex PER assay to commercial canned tuna samples was also demonstrated. The proposed test could be useful for detection of fraud and for seafood traceability.
Bottero
Maria Teresa
MT
Department of Animal Pathology, Faculty of Veterinary Medicine, University of Torino, via Leonardo da Vinci 44, 10095 Grugliasco, Torino, Italy. mariateresa.bottero@unito.it
Dalmasso
Alessandra
A
Cappelletti
Marco
M
Secchi
Camillo
C
Civera
Tiziana
T
D000818:Animals; D001483:Base Sequence; D045303:Cytochromes b / Q000235:genetics; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000235:genetics; D014644:Genetic Variation; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D014413:Tuna / Q000235:genetics*
D017931:DNA Primers; D004272:DNA, Mitochondrial; D045303:Cytochromes b