10.1021/acs.jafc.8b05652.s001
Ingunn A. Samdal
Ingunn A.
Samdal
Kjersti E. Løvberg
Kjersti E.
Løvberg
Anja B. Kristoffersen
Anja B.
Kristoffersen
Lyn R. Briggs
Lyn R.
Briggs
Jane Kilcoyne
Jane
Kilcoyne
Craig J. Forsyth
Craig J.
Forsyth
Christopher O. Miles
Christopher O.
Miles
A Practical ELISA for Azaspiracids in Shellfish via
Development of a New Plate-Coating Antigen
American Chemical Society
2019
Current methods
novel AZAs
AZA reference materials
LC-MS
food poisoning
New Plate-Coating Antigen Azaspiracids
Practical ELISA
AZA -3
Puget Sound
AZA -1-containing plate-coating conjugate
R 2
OVA
screening samples
ELISA method
2019-02-14 21:05:23
Journal contribution
https://acs.figshare.com/articles/journal_contribution/A_Practical_ELISA_for_Azaspiracids_in_Shellfish_via_Development_of_a_New_Plate-Coating_Antigen/7722149
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 μg/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 (<i>R</i><sup>2</sup> = 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.