Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 67, Issue 8, Pages 2369-2376Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.8b05652
Keywords
azaspiracid; AZA-1; ELISA; immunoassay; antibody; polyclonal; shellfish toxin; mussel
Funding
- project MARBioFEED under the First Call for Transnational Research Projects within the Marine Biotechnology ERA-NET [604814]
- Norwegian Research Council [139593/140]
- BIOTOX project - European Commission [514074]
- European Union Seventh Framework Programme (FP7/2007-2013) under the ECsafeSEAFOOD project [311820]
- Norwegian Veterinary Institute
- New Zealand Foundation for Research, Science and Technology (International Investment Opportunities Fund) [C10X0406]
- National Institute of Environmental Health Sciences (NIEHS), NIH, U.S.A. [ES10615]
- Marine Institute
- Marine Research Sub Programme of the National Development Plan 2007-2013
- European Regional Development Fund
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Azaspiracids (AZAs) are a group of biotoxins that appear periodically in shellfish and can cause food poisoning in humans. Current methods for quantifying the regulated AZAs are restricted to LC-MS but are not well suited to detecting novel and unregulated AZAs. An ELISA method for total AZAs in shellfish was reported recently, but unfortunately, it used relatively large amounts of the AZA-1-containing plate-coating conjugate, consuming significant amounts of pure AZA-1 per assay. Therefore, a new plate-coater, OVA-cdiAZA1 was produced, resulting in an ELISA with a working range of 0.30-4.1 ng/mL and a limit of quantification of 37 mu g/kg for AZA-1 in shellfish. This ELISA was nearly twice as sensitive as the previous ELISA while using 5-fold less plate-coater. The new ELISA displayed broad cross-reactivity toward AZAs, detecting all available quantitative AZA reference materials as well as the precursors to AZA-3 and AZA-6, and results from shellfish analyzed with the new ELISA showed excellent correlation (R-2 = 0.99) with total AZA-1-10 by LC-MS. The results suggest that the new ELISA is suitable for screening samples for total AZAs, even in cases where novel AZAs are present and regulated AZAs are absent, such as was reported recently from Puget Sound and the Bay of Naples.
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