DEVELOPMENT OF A CHEMILUMINESCENT ELISA FOR DETERMINING OKADAIC ACID IN SHELLFISH

ABSTRACT

An indirect competitive enzyme‐linked immunosorbent assay with chemiluminescent (CL‐ELISA) detection for okadaic acid (OA) in mussel muscle was developed. A hybridoma cell line secreting monoclonal antibody against OA was established after immunization of BALB/c mice with artificially synthesized OA‐bovine serum albumin as antigen. Luminol solution was used as the substrate for horseradish peroxidase. The detection limit was 0.175 ng/g. The range of average fortified recovery was 91.8–102.6% when OA was spiked in mussel muscle at levels of 50, 160 and 800 µg/kg. The standard curve for OA showed good linearity over the concentration range of 0.08125–20 ng/mL. The cross‐reactivity with dinophysistoxin1 and saxitoxin was 51.82 and 0%, respectively. In a residue study, the results obtained by CL‐ELISA correlated well within those obtained using the commercial ABRAXIS kit (ABRAXIS, Warminster, PA). The developed method is therefore suitable for detecting the residues of OA in shellfish.

PRACTICAL APPLICATIONS

Diarrhetic shellfish poisoning is a gastrointestinal syndrome that occurs in humans following the consumption of bivalve mollusks contaminated with OA with the main symptoms of diarrhea, nausea, vomiting and abdominal pain. OA widely exists at marine products and it is very important to develop a technique to monitor this toxin. In this study, a sensitive competitive indirect enzyme‐linked immunosorbent assay with chemiluminescence for determination of OA in mussel soft tissues was investigated. Chemiluminescent ELISA (CL‐ELISA) is a good alternative method for screening samples. This technique has the potential to improve the sensitivity of the immunoassays by at least two to three orders of magnitude compared with conventional colorimetric detection.