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Indirect enzyme-linked immunosorbent assay for saxitoxin in shellfish.

F S Chu, T S Fan

    Journal - Association of Official Analytical Chemists
    |January 1, 1985
    PubMed
    Summary

    A new indirect enzyme-linked immunosorbent assay (ELISA) detects saxitoxin (STX) in seafood. This assay shows high sensitivity and recovery rates for saxitoxin detection in shellfish.

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    Area of Science:

    • Analytical Chemistry
    • Immunology
    • Food Safety

    Background:

    • Saxitoxin (STX) is a potent marine neurotoxin responsible for paralytic shellfish poisoning.
    • Accurate and sensitive detection methods are crucial for monitoring STX levels in seafood to ensure public health.
    • Existing detection methods may have limitations in sensitivity, specificity, or practicality.

    Purpose of the Study:

    • To develop and validate an indirect enzyme-linked immunosorbent assay (ELISA) for the sensitive detection of saxitoxin (STX).
    • To characterize the specificity and cross-reactivity of the developed assay with related toxins.
    • To determine the assay's performance in detecting STX in real food matrices like shellfish.

    Main Methods:

    • Development of antibodies against STX through immunization of rabbits with STX-bovine serum albumin (STX-HCHO-BSA).
    • Coating microtiter plates with STX-HCHO-BSA or polylysine-STX for indirect ELISA.
    • Utilizing goat anti-rabbit IgG peroxidase conjugate and a chromogenic substrate for antibody detection.
    • Performing competitive indirect ELISA to assess cross-reactivity with other toxins.

    Main Results:

    • The developed indirect ELISA demonstrated high sensitivity for STX detection, with lower limits of detection at 25 pg/assay.
    • The assay showed minimal cross-reactivity with tetrodotoxin and carbamoyl-neo-STX-sulfate but cross-reacted with decarbamoyl-STX and neo-STX.
    • In shellfish (clams and mussels), the detection limit for STX was 50-100 ppb with recovery rates between 86.8-107%.

    Conclusions:

    • The indirect ELISA is a sensitive and specific method for detecting saxitoxin (STX).
    • This assay is suitable for monitoring STX contamination in shellfish, contributing to food safety.
    • The method offers reliable quantification of STX in complex food matrices.

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