Application of Relative Quantification TaqMan Real-Time Polymerase Chain Reaction Technology for the Identification and Quantification of Thunnus alalunga and Thunnus albacares

Abstract

A novel one-step methodology based on real-time Polymerase Chain Reaction (PCR) technology has been developed for the identification of two of the most valuable tuna species. Nowadays, species identification of seafood products has a major concern due to the importing to Europe of new species from other countries. To achieve this aim, two specific TaqMan systems were devised to identify Thunnus alalunga and Thunnus albacares. Another system specific to Scombroidei species was devised as a consensus system. In addition, a relative quantification methodology was carried out to quantify T. alalunga and T. albacares in mixtures after the relative amount of the target was compared with the consensus. This relative quantification methodology does not require a known amount of standard, allowing the analysis of many more samples together and saving costs and time. The utilization of real-time PCR does not require sample handling, preventing contamination and resulting in much faster and higher throughput results.

Novel approach for the simultaneous detection of DNA from different fish species based on a nuclear target: quantification potential

“…Previously reported methods based on PCR or real-time PCR for fish in different samples focused on the identification of specific species [1,[25][26][27][28] or on the use of mtDNA as a The criteria [23] we as follows: slope between -3.6 and -3.1; efficiency between 0.9 and 1.1; coefficient of correlation (R 2 ) between 0.99 and 0.999.…”

Comparative study of DNA extraction methods from fresh and processed yellowfin tuna muscle tissue

“…[7,8] Nevertheless, fragment size is a limiting factor for the subsequent PCR reaction, which is based on the selective amplification of specific regions of DNA using oligonucleotides. [8] PCR [9][10][11] and its modifications-polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), [12][13][14][15] Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP), [16,17] real-time PCR, [18][19][20][21] and Polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA) [22] -represent crucial approaches for tuna fish species identification. These approaches comprise DNA extraction from the sample, PCR, and electrophoresis, or alternatively other detection systems for the evaluation of the final results.…”

Aquatic Food

“…In another study, the mitochondrial 16S RNA gene was amplified by TaqMan PCR systems to quantify the relative amounts of two tuna species in mixtures of raw muscle; the results were promising for raw fish but became highly inaccurate in case of canned samples (Lopez and Pardo, 2005).…”