Development and validation of a maleimide-based enzyme-linked immunosorbent assay for the detection of tetrodotoxin in oysters and mussels
Elliott, Christopher T.
Klijnstra, Mirjam D.
The recent detection of tetrodotoxins (TTXs) in puffer fish and shellfish in Europe highlights the necessity to monitor the levels of TTXs in seafood by rapid, specific, sensitive and reliable methods in order to protect human consumers. A previous immunoassay for TTX detection in puffer fish, based on the use of self-assembled monolayers (SAMs) for the immobilization of TTX on maleimide plates (mELISA), has been modified and adapted to the analysis of oyster and mussel samples. Changing dithiol for cysteamine-based SAMs enabled reductions in the assay time and cost, while maintaining the sensitivity of the assay. The mELISA showed high selectivity for TTX since the antibody did not cross-react with co-occurring paralytic shellfish poisoning (PSP) toxins and no interferences were observed from arginine (Arg). Moreover, TTX-coated maleimide plates stored for 3 months at −20 °C and 4 °C were stable, thus when pre-prepared, the time to perform the assay is reduced. When analyzing shellfish samples, matrix effects and toxin recovery values strongly depended on the shellfish type and the sample treatment. Blank oyster extracts could be directly analyzed without solid-phase extraction (SPE) clean-up, whereas blank mussel extracts showed strong matrix effects and SPE and subsequent solvent evaporation were required for removal. However, the SPE clean-up and evaporation resulted in toxin loss. Toxin recovery values were taken as correction factors (CFs) and were applied to the quantification of TTX contents in the analysis of naturally-contaminated shellfish samples by mELISA. The lowest effective limits of detection (eLODs) were about 20 and 50 µg/kg for oyster extracts without and with SPE clean-up, respectively, and about 30 µg/kg for mussel extracts with both protocols, all of them substantially below the eLOD attained in the previous mELISA for puffer fish (230 µg/kg). Analysis of naturally-contaminated samples by mELISA and comparison with LC-MS/MS quantifications demonstrated the viability of the approach. This mELISA is a selective and sensitive tool for the rapid detection of TTX in oyster and mussel samples showing promise to be implemented in routine monitoring programs to protect human health.
63 - Agricultura. Silvicultura. Zootècnia. Caça. Pesca
Is part of
Reverté, Laia, Maria Rambla-Alegre, Sandra Leonardo, Carlos Bellés, Katrina Campbell, Christopher T. Elliott, Arjen Gerssen, Mirjam D. Klijnstra, Jorge Diogène, and Mònica Campàs. 2018. "Development And Validation Of A Maleimide-Based Enzyme-Linked Immunosorbent Assay For The Detection Of Tetrodotoxin In Oysters And Mussels". Talanta 176: 659-666. Elsevier BV. doi:10.1016/j.talanta.2017.08.058
Grant agreement number
INIA/Programa Estatal de promoción del talento y su empleabilidad en I+D+I/RTA2013-00096-00-00/ES/Promoción de una acuicultura de bivalvos y pesqueria sostenible y segura mediante modelizacióin y caracterización del riesgo de toxinas emergentes/PROMAQUA
Aigües Marines i Continentals
This item appears in the following Collection(s)
- ARTICLES CIENTÍFICS 
The following license files are associated with this item:
Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by-nc-nd/4.0/