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Domoic Acid

The Severity of Environmental Contamination Caused by Domoic Acid

Domoic acid (DA) is produced by the metabolism of certain species of diatoms in the genus Pseudo-nitzschia and Nitzschia. When these algae multiply and form red tides, a large amount of domoic acid will be produced in the initial stage after the formation of red tides, which will pollute the seawater environment. DA accumulates through the food chain. Humans can cause poisoning after eating seafood contaminated by DA. Domoic acid was found in shellfish samples from North America, New Zealand, Japan, Denmark, Scotland, France, Spain, Portugal, and other regions. Hazardous incidents caused by domoic acid on the coasts of Canada and the United States have been continuously reported.

The Necessity of Monitoring Domoic Acid with ELISA Testing

Fig. 1 The chemical structure of domoic acid

The structure of domoic acid is similar to that of kainic acid and glutamic acid, and it is an agonist of kainate receptors. The pure product is a white solid powder, soluble in water, and will not degrade in alkaline solutions at room temperature or light. Domoic acid and glutamic acid (Glu) as a neurotransmitter have the same function of causing nerve cell excitement, but its strength is more than 100 times higher than Glu and 2-3 times higher than kainic acid (KA). DA can enter the brain through the high-affinity transport system of the blood-brain barrier. After poisoning, the human body will experience nausea, vomiting, and gastritis. In severe cases, it can cause dizziness, visual disturbance, memory loss, and coma. DA has been recognized since the Canadian poisoning incident in 1987, and the U.S. Food and Drug Administration (FDA) has identified it as one of the four major marine biological toxins. Before and after the occurrence of red tides and blooms, the detection of domoic acid should be strengthened. Given that ELISA is sensitive, specific, and does not require special equipment, it has been widely used in the detection of domoic acid.

ELISA Type

Indirect Competitive ELISA

The Advantages of ELISA Testing

Easy to use
Lower cost
High-throughput detection and analysis
High sensitivity and specificity

ELISA Procedure for Domoic Acid Testing

The optimal concentrations of the DA-OVA antigen was coated in the ELISA micro well plates in triplicates and incubated overnight at 4°C.

Washed three times by PBST and by PBS subsequently, blocked by 5% PBSM (200 µL/well), and kept at 37°C for 2 h.

Washed with PBST and PBS respectively, equivalent volume of the 1C3 mAb and the free DA toxin was reacted at various concentrations (0, 0.01, 0.05, 0.1, 1, 5, 10, 100, 1000 ng/mL) in triplicates. After proper mixing, incubated at 37°C for 30 min.

Later, the reaction mix was transferred to the ELISA plates (100 µL/well) with each standard concentration and incubated at 37°C for 1 h.

Terminated by the addition of 50 µL/well of 2M H₂SO₄, the absorbance was measured at 450 nm by microplate reader.

Creative Diagnostics has been committed to algal toxins testing by ELISA. Supported by rich professional knowledge and diversified ELISA kits products, we provide high-quality customized ELISA kits services, professional ELISA testing services, and ELISA development services related to the detection of domoic acid. If you wish a lot of careful data, please contact us.

References

Weirich, C.A,; Miller, T.R. Freshwater harmful algal blooms: toxins and children's health. Curr Probl Pediatr Adolesc Health Care. 2014, 44(1): 2-24.
Saeed, A.F.; et al. Domoic acid: Attributes, exposure risks, innovative detection techniques and therapeutics. Algal Res. 2017, 24: 97–110.
Ling, S.; et al. The Preparation and Identification of a Monoclonal Antibody against Domoic Acid and Establishment of Detection by Indirect Competitive ELISA. Toxins (Basel). 2017, 9(8): 250.

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