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Development and validation of an enzyme‐linked immunosorbent assay (ELISA) for the detection of ciguatoxin using chicken immunoglobulin Y as an analytic tool
Author(s) -
EmpeyCampora Cara,
Hokama Yoshitsugi,
Yabusaki Kenichi,
Isobe Minoru
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a1140-b
Subject(s) - ciguatoxin , monoclonal antibody , chemistry , antibody , enzyme , microbiology and biotechnology , fish <actinopterygii> , chromatography , biochemistry , biology , immunology , fishery
A sandwich enzyme‐linked immunosorbent assay (ELISA) was developed for the detection of ciguatoxin (CTX) using a chicken immunoglobulin Y (IgY) to the synthetically produced ABCD region of the ciguatoxin molecule and a previously produced mouse anti‐CTX monoclonal antibody shown to react with the JKLM region. Ciguatoxin from fish extract positively characterized for CTX was sandwiched between purified IgY adhered onto a solid‐phase and the mouse anti‐CTX IgG. The assay was validated by determination of sensitivity, working range, cross reactivity, accuracy, precision, and reproducibility. Evaluation of the usefulness of the assay to screen and positively identify ciguatoxic wild caught fish is currently underway. [Supported, in part, by the Hokama‐Yagawa Fund, University of Hawaii Foundation]