Detection of Transgenic Atlantic and Coho Salmon by Real-time PCR
- 206 Downloads
Genetic modifications (GM) have been applied to salmon to generate fast-growing strains for potential use in aquaculture. In November 2015, the first transgenic salmon (AquAdvantage® Atlantic salmon) was accepted for commercialization in the USA under defined conditions. The presence of GM food products in the marketplace stimulates the need for detection methods to allow screening for the presence of genetic modifications in seafood products. This paper first shows that it is possible to obtain amplifiable DNA from raw and processed products containing salmon. Detection methods by real-time PCR are proposed in this work. An endogenous gene target was designed to detect salmonid species DNA in samples. In addition, detection methods using real-time PCR were developed for two GM salmon possessing growth hormone transgenes: the AquAdvantage® Atlantic salmon (Salmo salar) developed by AquaBounty for commercial purposes, and the coho salmon (Oncorhynchus kisutch) developed for research purposes by Fisheries and Oceans Canada. The methods are able to detect at least 20 copies of the target. It was found however that one of the construct-specific methods for the AquAdvantage® salmon detection did not work on AquAdvantage® genomic DNA even though it works on the sequence published in GenBank. The other assay however was found to reliably detect AquAdvantage® transgenic sequences in genomic DNA.
KeywordsSalmon Transgenic Detection AquAdvantage Real-time PCR
This research was conducted within a Belgian research project (Convention RF 11/6242 UGMMONITOR) financed by the Belgian Federal Public Service for Public Health, Food Chain Safety and Environment. We also thank Cécile Ancion, Denis Roulez, and Gaëlle Antoine from the GMO team of CRA-W.
Compliance with Ethic Requirements
Conflict of Interest
Frédéric Debode declares that he has no conflict of interest. Eric Janssen declares that he has no conflict of interest. Aline Marien declares that she has no conflict of interest. Robert H. Devlin declares that he has no conflict of interest. Kathrin Lieske declares that she has no conflict of interest. Joachim Mankertz declares that he has no conflict of interest. Gilbert Berben declares that he has no conflict of interest.
This article does not contain studies with human or animal subjects. Samples used are food samples.
- AFNOR XP V03-020-2 (2008). Produits alimentaires. Détection et quantification des organismes végétaux génétiquement modifies et produits dérivés. Partie 2: méthodes basées sur la réaction de polymérisation en chaîne. Norme expérimentaleGoogle Scholar
- AquaBounty Technologies, Inc, 2010, An Atlantic salmon (Salmo salar L.) bearing a single copy of the stably integrated α-form of the opAFP-GHc2 gene construct at the α- locus in the EO-1α line, Environmental Assessment for AquAdvantage® Salmon. http://www.fda.gov/downloads/AdvisoryCommittees/CommitteesMeetingMaterials/VeterinaryMedicineAdvisoryCommittee/UCM224760.pdf (last consulted on 18 June 2016)
- Cowx IG, Bolland JD, Nunn AD, Kerins G, Stein J, Blackburn J, Hart A, Henry C, Britton J, Capp G, Peeler E (2010) Defining environmental risk assessment criteria for genetically-modified fishes to be placed on the EU market. EFSA Support Publication 7Google Scholar
- Debode F, Marien A, Janssen E, Bragard C, Berben G (2017) The influence of amplicon length on real-time PCR results. Biotechnol Agron Soc Environ 21:3–11Google Scholar
- FAO (2014) The state of world fisheries and aquaculture. Rome, 223pp. ISBN 978-92-5-108275-1Google Scholar
- FDA (2015) AquAdvantage® salmon approval letter and appendix. http://www.fda.gov/AnimalVeterinary/DevelopmentApprovalProcess/GeneticEngineering/GeneticallyEngineeredAnimals/ucm466214.htm (last consulted on 22 June 2016)
- Forsberg CW, Phillips JP, Golovan SP, Fan MZ, Meidinger RG, Ajakaiye A, Hilborn D, Hacker RR (2003) The enviropig physiology, performance, and contribution to nutrient management advances in a regulated environment: the leading edge of change in the pork industry. J Anim Sci 81:E68–E77CrossRefGoogle Scholar
- ISO 21571 (2005) Foodstuffs. Methods of analysis for the detection of genetically modified organisms and derived products. Nucleic acid extraction. International Organization for Standardization, GenevaGoogle Scholar
- McLeod C, Grice J, Campbell H, Herleth T (2006) Super salmon: the industrialisation of fish farming and the drive towards GM technologies in salmon production. http://sro.sussex.ac.uk/id/eprint/37927
- Menozzi D, Mora C, Merigo A (2013) Genetically modified salmon for dinner? Transgenic salmon marketing scenarios. AgBioforum 15:276–293Google Scholar
- Ohno S, Muramoto J, Klein J, Atkin NB (1969) Diploid-tetraploid relationship in clupeoid and salmonoid fish. Chromosomes Today 2:139–147Google Scholar
- Phillips JP, Golovan SP, Meidinger RG, Forsberg CW (2006) Transgenic enhancement of nutrient cycling: moving toward an environmentally sustainable animal agriculture. In Proceedings of the 8th World Congress on Genetics Applied to Livestock Production, Belo Horizonte, Minas Gerais, Brazil, 13–18 August, 2006. Instituto Prociência, pp 19–02Google Scholar
- Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar