Detection of ornamental transgenic fish by real-time PCR and fluorescence microscopy

Numbers of ornamental transgenic fish are increasing, and some of them are illegally imported into Europe. The fish are modified to display different fluorescent colors under UV light. In this article, we propose real-time PCR methods to detect fish presenting green, yellow and red fluorescent coloring. The methods were tested with success and were able to detect illegally imported fish in two European countries. The article also discusses some practical information that can be useful for routine analysis. In addition, a real-time PCR test able to highlight the presence of fish DNA in general is proposed to check the amplifiability of the DNA extracted from common bony fish species of the teleost subclass. Finally, as the testing by PCR can take several days and rapid decisions must be taken with living organisms, we explored the potential of fluorescence microscopy as a screening test to determine whether animals are suspect or can be released.