Development of a Sandwich Enzyme-Linked Immunosorbent Assay for Detection and Quantification of Clam Residues in Food Products.

Stef J Koppelman,Ashley L Lardizabal,Steve L Taylor,Lynn Niemann,Joe L Baumert,Erika Di Zazzo

doi:10.1155/2021/6685575

Stef J Koppelman, Ashley L Lardizabal + Show 4 more

Open Access

https://doi.org/10.1155/2021/6685575

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Abstract

Seafood is a frequent cause of allergic reactions to food globally. The presence of undeclared trace amounts of clam can cause allergic reactions in sensitive individuals. Limited tools are available to test food products for the presence of traces of clam. We report on the development of a sandwich ELISA that can detect and quantify clam protein in food. Antisera against a mix of two commercially important clam species, Atlantic Surf (Spisula solidissima) and ocean quahog (Arctica islandica), were raised in rabbit and sheep. A sandwich ELISA was constructed with this antisera, and sensitivity and specificity were evaluated. Also, model food products spiked with clam protein were analyzed to assess the performance of the ELISA. Comparison was made with a commercially available ELISA for crustacea. The lower limit of quantification of the sandwich ELISA is 2.5 ppm clam protein in food samples, allowing the detection of low amounts of clam that may trigger a reaction in clam allergic patients. The sandwich ELISA was highly specific with cross-reactivity only noted for other molluscan shellfish (mussel and scallop). Clam protein in tomato juice and potato cream soup was detected well with recoveries ranging from 65 to 74% and from 74 to 113%, respectively. However when potato cream soup was retorted, the recover fell to 20%, imposing the risk of underestimating the clam content of a food product. A commercially available crustacean ELISA test was not suitable to detect clam protein. The sandwich ELISA described here is suitable for detection and quantification of clam protein in food products. Care should be taken with food products that have been retorted as the results may be underestimated.

Highlights

Clams are molluscan shellfish, part of the second largest phylum in the animal kingdom having 104 edible species according to the Food & Agriculture Organization of the United Nations
Molluscan shellfish are further subdivided into 9-10 classes with three classes that are commonly ingested—bivalves, cephalopods, and gastropods
We report the development of a sensitive and specific Enzyme-linked immunosorbent assay (ELISA) for the detection of clam residues in foods, based on the use of polyclonal antisera raised against a mixture of processed clam proteins

Summary

Introduction

Part of the second largest phylum in the animal kingdom having 104 edible species according to the Food & Agriculture Organization of the United Nations. In the US, the production of ocean quahog clams was 11.3 million pounds in 2019, while the production of Atlantic Surf clams was 37 million pounds in 2017 (http://www.fishwatch.gov). The Northern quahog or hard clam (Mercenaria mercenaria) is another commercial species in the US but annual production of Northern quahog is substantially lower than that of ocean quahog and Atlantic Surf clam (http://www.fishwatch.gov). Both molluscan and crustacean shellfish are well-known causes of IgE-mediated, immediate hypersensitivity allergic reactions [1, 2]. In a nationwide survey of 30,018 individuals in Taiwan, 1.2% reported allergies to molluscan shellfish [13] but clinical confirmation was not sought. We report the development of a sensitive and specific ELISA for the detection of clam residues in foods, based on the use of polyclonal antisera raised against a mixture of processed clam proteins

Materials and Methods

Results and Discussion

Sandwich ELISA for Clam Protein

Conclusion

Conflicts of Interest