Eliska Servusova
Veterinary Research Institute, Czech republic
Title: Identification of selected tuna species in commercial products by real-time PCR
Biography
Biography: Eliska Servusova
Abstract
In this study, systems for the identification of four tuna species (skipjack tuna Katsuwonus pelamis, yellowfin tuna Thunnus albacares, bullet tuna Auxis sp. and atlantic bonito Sarda sp.) were developed. First, raw samples of these species and an internal control mixture confirming the fish muscle of the species Thunnus sp. Raw muscle, mixture and samples were isolated with DNeasy mericon Food Kit (Qiagen GmbH, Germany). The raw samples were assayed for DNA concentration and purity using a spectrophotometer. Primers and probe sequences were specifically designed to identify selected species. Next, primers and a probe for the endogenous 12S rRNA gene were designed to determine the presence of amplifiable fish (especially tuna) DNA in the sample. The species specificity of the designed primers and probes on DNA samples of various tuna and bonito species was also verified. Detection limit for selected species and determination coefficient R2 and real-time PCR test were determined. To evaluate the developed real-time PCR methods, 70 commercial tuna products were analyzed. These commercial products were acquired on local markets in the Czech Republic. Most often, they were canned in their own juice, oil, with various ingredients, spread, pate, various tuna salads and tuna dishes. The sample range also included three pet food products.