SureFood® FISH ID – Novel analytical tools for rapid fish species identification Lysann Hinz1, Dr. Janine Beutlich1, Patrizia Eckelt1, Anja Palisch1, Dr. Martin Mehl2, Dr. Steffen Mergemeier1 1 CONGEN Biotechnologie GmbH, Robert-Roessle-Str. 10, 13125 Berlin, Germany; Tel: +49 (0) 30 9489 3500 E-mail: info@congen.de 2 R-Biopharm AG, An der neuen Bergstr. 17, 64297 Darmstadt, Germany; Tel: +49 (0) 6151 8102-0, E-mail: info@r-biopharm.de Scope and Purpose According to EU regulation 1379/2013 on the common organization of the markets of fishery and aquaculture products, fish products must be mandatory labeled with the commercial designation and the scientific species name since 13th December 2014. Therefore, specific detection methods are required. For a quick and reliable on-site application novel real-time PCR based test systems were developed. Results Analyses showed that SureFood® FISH ID could successfully differentiate between presently 11 different fish species. Amplification curves are exemplary shown for Oncorhynchus nerka (Figure 2) and Gadus morhua (Figure 3). Figure 2: Amplification curves generated by the SureFood® FISH ID Oncorhynchus nerka IAAC system (ABI 7500). (A) Specific detection of O. nerka in the FAM-channel. (B) Animal DNA and IAC detection in the VIC-channel. (A) (B) Figure 3: Amplification curves generated by the SureFood® FISH ID Gadus morhua IAAC system (Bio-Rad CFX96). (A) Specific detection of G. morhua in the FAM-channel. (B) Animal DNA and IAC detection in the VIC-channel. O. nerka (100%) Positive template control O. nerka (1%) Negative template control G. morhua (100%) G. morhua (1%) Figure 1: Fish species that can be identified by SureFood® FISH ID. Methodology The real-time PCR based SureFood® FISH ID product line offers easy to handle tools that guarantee species differentiation and evaluation of results within three hours for a broad range of commercially interesting fish species (Figure 1). For DNA-preparation of raw material the use of SureFood® PREP Basic and for highly processed food the use of SureFood® PREP Advanced (both CONGEN) is recommended. Each reaction contains an internal amplification control (IAC) and an internal detection assay for animal DNA (IAAC). The methods are developed for the detection of specific fish DNA in muscle meat mixture at a relative amount of 1-2%. Detection of the respective fish species is performed in the FAM-channel, while animal DNA and IAC detection is run in the VIC-channel. Assay validation was performed on commonly used real-time PCR devices (e.g. Roche LightCycler 2.0, Roche LightCycler 480 II, Qiagen Rotor-Gene Q, Applied Biosystems 7500, Bio-Rad CFX96, Agilent Mx3005P). Conclusion To our knowledge SureFood® FISH ID offers the fastest method for fish species identification currently available on the market and provides an easy to handle tool to fulfill consumer information requirements under the terms of EU 1379/2013.