FN Archimer Export Format
PT J
TI Fluorescent Receptor Binding Assay for Detecting Ciguatoxins in Fish
BT 
AF HARDISON, D. Ransom
   HOLLAND, William C.
   MCCALL, Jennifer R.
   BOURDELAIS, Andrea J.
   BADEN, Daniel G.
   DARIUS, H. Taiana
   CHINAIN, Mireille
   TESTER, Patricia A.
   SHEA, Damian
   QUINTANA, Harold A. Flores
   MORRIS, James A., Jr.
   LITAKER, R. Wayne
AS 1:1;2:1;3:2,3;4:2;5:2;6:4;7:4;8:1,5;9:6;10:7;11:1;12:1;
FF 1:;2:;3:;4:;5:;6:;7:;8:;9:;10:;11:;12:;
C1 NOAA, Ctr Coastal Fisheries & Habitat Res, Beaufort, NC USA.
   Univ N Carolina, MARBIONC CREST Res Pk, Wilmington, NC 28401 USA.
   SeaTox Res Inc, UNCW CREST Res Pk, Wilmington, NC USA.
   ILM, UMR EIO 241, Lab Tox Microalgae, Papeete, Tahiti, France.
   JHT Inc, Orlando, FL USA.
   N Carolina State Univ, Environm Chem & Toxicol Lab, Raleigh, NC 27695 USA.
   US FDA, Div Seafood Sci & Technol, Gulf Coast Seafood Lab, Dauphin Isl, AL USA.
C2 NOAA, USA
   UNIV N CAROLINA, USA
   SEATOX RES INC, USA
   INST LOUIS MALARDE, FRANCE
   JHT INC, USA
   UNIV N CAROLINA STATE, USA
   US FDA, USA
UM EIO
IN DOAJ
IF 2.806
TC 57
UR https://archimer.ifremer.fr/doc/00664/77575/79555.pdf
LA English
DT Article
AB Ciguatera fish poisoning is an illness suffered by > 50,000 people yearly after consumption of fish containing ciguatoxins (CTXs). One of the current methodologies to detect ciguatoxins in fish is a radiolabeled receptor binding assay (RBA((R))). However, the license requirements and regulations pertaining to radioisotope utilization can limit the applicability of the RBA((R)) in certain labs. A fluorescence based receptor binding assay (RBA((F))) was developed to provide an alternative method of screening fish samples for CTXs in facilities not certified to use radioisotopes. The new assay is based on competition binding between CTXs and fluorescently labeled brevetoxin-2 (BODIPY (R)-PbTx-2) for voltage-gated sodium channel receptors at site 5 instead of a radiolabeled brevetoxin. Responses were linear in fish tissues spiked from 0.1 to 1.0 ppb with Pacific ciguatoxin-3C (P-CTX-3C) with a detection limit of 0.075 ppb. Carribean ciguatoxins were confirmed in Caribbean fish by LC-MS/MS analysis of the regional biomarker (C-CTX-1). Fish (N = 61) of six different species were screened using the RBA((F)). Results for corresponding samples analyzed using the neuroblastoma cell-based assay (CBA-N2a) correlated well (R-2 = 0.71) with those of the RBA((F)), given the low levels of CTX present in positive fish. Data analyses also showed the resulting toxicity levels of P-CTX-3C equivalents determined by CBA-N2a were consistently lower than the RBA((F)) affinities expressed as% binding equivalents, indicating that a given amount of toxin bound to the site 5 receptors translates into corresponding lower cytotoxicity. Consequently, the RBA((F)), which takes approximately two hours to perform, provides a generous estimate relative to the widely used CBA-N2a which requires 2.5 days to complete. Other RBA((F)) advantages include the long-term(> 5 years) stability of the BODIPY (R)-PbTx-2 and having similar results as the commonly used RBA((R)). The RBA((F)) is cost-effective, allows high sample throughput, and is well-suited for routine CTX monitoring programs. 
PY 2016
PD APR
SO Plos One
SN 1932-6203
PU Public Library Science
VL 11
IS 4
UT 000374131200077
DI 10.1371/journal.pone.0153348
ID 77575
ER

EF