A handheld sensor assay for the identification of grouper as a safeguard against seafood mislabeling fraud

Increases in international trade and global seafood consumption, along with fluctuations in the supply of different seafood species, have resulted in fraudulent product mislabeling. Grouper species, due to their high demand and varied commercial availability, are common targets for mislabeling by exploiting inefficient inspection practices. Compounding this problem is the fact that there are currently 64 species of fish from eleven different genera allowed to be labeled “grouper” per U.S. Food and Drug Administration guidelines. This wide diversity makes it difficult for regulators to discern legally salable groupers from restricted species. To obviate taxonomic misidentification when relying on external phenotypic characteristics, regulatory agencies are now employing genetic authentication methods which typically offer species-level resolution. However, standard genetic methods such as DNA barcoding require technical expertise and long turnover times, and the required instrumentation is not amenable for on-site analysis of seafood. To obviate some of these limitations, we have developed a handheld genetic sensor that employs a real-time nucleic acid sequence-based amplification assay (RT-NASBA) previously devised in our lab, for the analysis of fish tissue in the field. The base RT-NASBA assay was validated using a lab-based, benchtop RNA purification method as well as non-portable, commercial RT-NASBA analyzer. Described herein, is an uncomplicated method for purifying RNA from fish tissue in the field, which had similar efficiency to the benchtop method demonstrated through direct comparisons. We have also demonstrated that the field sensor is only slightly less sensitive than the benchtop instrument, and could discern 80.3% of groupers (no target sequence available for three species) on the 2014 FDA Seafood List from potential impostors. The complete field assay requires fewer than 80 min for completion and can be performed outside of the lab in its entirety.