Application Guide - QIAxcel Advanced

130 QIAxcel Advanced Application Guide 10/2016 Species determination for meat using PCR-RFLP analysis on the QIAxcel® system Christoph Graf,1 Agne Rüegg-Kuslyte,2 and Roger Kuhn2 1 Official Regulatory Authority for Food of Canton Bern, Bern, Switzerland 2 Department of Life Science and Facility Management, Zurich University of Applied Sciences (ZHAW), Waedenswil, Switzerland A strategy using PCR followed by restriction fragment length polymorphism analysis (PCR-RFLP) is used to identify meat from various animal species in food. Analysis results from the QIAxcel system and from agarose gel electrophoresis were compared for 14 exotic and game species. Although both techniques were suitable for species differentiation, the use of the QIAxcel was less time consuming and provided the advantage of electronic documentation. Introduction Today, meats from exotic and game animals from all around the world provide an attractive alternative for people looking for unforgettable sensory experiences. Falsification of specialty meat is very common due to the tremendous profit gained by selling less costly meat labeled as meat from a more expensive species. A method that is sensitive enough to detect the small but relevant difference between meat from specific species is essential for official food control authorities to verify claims made about specialty meat. Protein-based methods for species identification, such as isoelectric focusing (IEF) or immunological methods, are not adequate because the soluble muscle proteins in processed meat products (heated or marinated) are rapidly and efficiently degraded. Nucleic acid-based analytical methods for the differentiation and identification of animal species in food have been commonly used in the last 20 years (1, 2). In this study, PCR amplification of the cytochrome b (cytb) gene followed by restriction fragment length polymorphism analysis (PCR-RFLP) was used to differentiate 14 different exotic or game species. Traditionally, gel electrophoresis has been used to detect PCR-RFLP DNA fragments. However, this method is laborious, time consuming, and hazardous due to the use of ethidium bromide or similar intercalating dyes that are mutagenic and dangerous for human health. In addition, gel data can not be used directly for publication or archiving. As an alternative method for discriminating animal species in processed food and meat, we evaluated the QIAxcel capillary electrophoresis system, a computer-controlled system that provides electronic documentation.

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