DEVELOPMENT OF AN IMMUNOENZYME METHOD FOR THE QUANTITATIVE DETERMINATION OF DOMOIC ACID IN SEAFOODS

DOI: https://doi.org/None

N.V. Kochkina (1), P.M. Shalimov (1), A.V. Malinin (1), V.V. Vrublevskaya (2), O.S. Morenkov (2) 1 -Scientific Centre «Signal», Bolshaya Olenya Street, 8, Moscow, Russian Federation, 107014; 2 -Institute of Cell Biophysics, Russian Academy of Sciences, Institutskaya Street, Pushchino, 3, Moscow Region, Russian Federation, 142290

Introduction. Amnesic shellfish poisoning is caused by consumption of seafoods contaminated with domoic acid (DA) and is accompanied by gastric colics, nausea, convulsions, hallucinations, loss of orientation and amnesia. As a preventive measure against these poisonings, European Union Directive 2002/226/ЕСand Russian Federation Sanitary Regulations and Norms SanPiN2.3.2.2401-08 prescribe a mandatory check of seafoods for the occurrence of DA in them. Enzyme-linked immunosorbent assay (ELISA) is one of the most promising DA detection methods; it combines a high sensitivity, specificify, simplicity and assaying speed. No Russian-made domoic acid ELISA assays are available. Aim of the study. To develop a sensitive and specific ELISA assay for detection and quantitative determination of DA in seafoods. Results. A direct competitive ELISA assay based on obtained DA-specific polyclonal antibodies for DA determination was developed. The threshold concentration of the toxin revealed by the developed assay was 0,05–0,07 ng/ml; the 50% inhibitory concentration, 0,7–1,0 ng/ml; the assay range, 0,1–5,0 ng/ml sample. The developed ELISA assay enabled a reliable detection of the toxin in shellfish tissue samples at its content from 5 ng/ml, which is 4000 times as low as the maximum permissible value for DA (20 μg/ml). Conclusion. The developed ELISA assay can be used to monitor contamination of seafoods with domoic acid as a preventive measure against amnesic shellfish poisoning.
Keywords: 
Key words. Amnesic shellfish poisoning, domoic acid, enzyme-linked immunosorbent assay
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