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Ciguatoxin

The Severity of Food Contamination Caused by Ciguatoxin

Ciguatoxin (CTX) is one of the most dangerous red tide biotoxins. Its main source of toxin production is the toxic Gambierdiscus toxicus, Prorocentrum lima, Pyrocystis, and subtropical benthic microalgae species. When coral fish eat toxic algae, they can accumulate in the fish. It is harmless to the fish itself. It is transmitted and enriched through the food chain and becomes a more toxic toxin after biological oxidation and metabolism. Since toxins will be enriched through the food chain, the larger the fish, the higher the toxins it contains. Three types of ciguatoxin have been found, namely Pacific ciguatoxin, Caribbean ciguatoxin, and Indian ciguatoxin. In terms of quantity and toxicity, ciguatoxin is one of the most toxic toxins known to mammals. There is evidence that due to the migration of fish and the expansion of trade in fish products, ciguatoxin has affected many areas and has become a major obstacle to fishery economic development and public health.

The Necessity of Monitoring Ciguatoxin with ELISA

The Necessity of Monitoring Ciguatoxin with ELISA Fig. 1 The chemical structures of the
four major ciguatoxin congeners (Tsumuraya, T.; Hirama, M., 2019)

CTX is a fat-soluble high-ether substance, colorless, odorless, insoluble in water, heat-resistant, and not easily destroyed by gastric acid. Ciguatoxin is not dangerous to the fish themselves. The toxins will accumulate slowly. The harm caused by ciguatoxin to the human body is mainly caused by eating fish containing ciguatoxin. The toxin has been isolated from more than 400 kinds of fish, causing a huge safety hazard to human food. According to reports, the average number of ciguatoxin poisonings that occur every year exceeds 50,000. In Hawaii and Florida, ciguatoxin has even affected the development of the local fishing industry. In order to ensure food safety, prevent ciguatoxin from causing human poisoning, and monitor the pollution of ciguatoxin to local fisheries, as a fast and efficient immunoassay method, ELISA has long been used to detect the content of ciguatoxin in fish products.

ELISA Types

Sandwich ELISA

The Advantages of ELISA Testing

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

ELISA Procedure for Ciguatoxin Testing

The microtiter plates were coated with 100 µL of a mixture of mAbs 3G8 (10 µg/mL) and 10C9 (10 µg/mL) in PBS, and incubated overnight at 4°C.

After the mAb solution wad decanted, the blocking buffer (400 µL/well) was added to each well and incubated for 1 h at room temperature.

After removed the blocking buffer, serially diluted solutions (100 µL) of CTX in D1 buffer were added to the plate and incubated for 1 h at RT.

After washing three times, the solution (100 µL) of ALP-conjugated 8H4 (2 µg/mL) dissolved in D2 buffer was added to wells and incubated for 1 h.

After washing three times, 100 µL of a solution of AP Fluorescent Substrate System was added to each well.

Measure the fluorescence intensity (excitation, λ=435 nm; emission, λ=555 nm) with a microtiter plate reader after standing for 15-30 min.

Creative Diagnostics has been committed to food biotoxins testing by ELISA. We provide a variety of biotoxins detection ELISA kits. 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 ciguatoxin. If you wish a lot of careful data, please contact us.

References

Tsumuraya, T.; Hirama, M. Rationally Designed Synthetic Haptens to Generate Anti-Ciguatoxin Monoclonal Antibodies, and Development of a Practical Sandwich ELISA to Detect Ciguatoxin. Toxins, 2019, 11(9): 533.
Tsumuraya, T.; et al. Highly Sensitive and Practical Fluorescent Sandwich ELISA for Ciguatoxin. Anal Chem. 2018, 90(12): 7318-7324.

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