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. 2021 Mar 19:2021:6685575.
doi: 10.1155/2021/6685575. eCollection 2021.

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

Stef J Koppelman  1 , Ashley L Lardizabal  1 , Lynn Niemann  1 , Joe L Baumert  1 , Steve L Taylor  1
Affiliations

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

Stef J Koppelman et al. Biomed Res Int. .

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.

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Conflict of interest statement

None of the authors report conflict of interest related to the present work.

Figures

Figure 1
Figure 1
Electrophoretic pattern of clam protein extracts and immunoreactivity of clam antisera. (a) SDS-PAGE with Coomassie staining. (b) Immunoblot with sheep serum. (c) Immunoblot with rabbit serum. Mix (Ret.): mix of Atlantic Surf clam and ocean quahog calm, retorted; Surf: Atlantic Surf clam; Quahog: quahog clam; MWs: molecular weight markers (indicated in kDa at right margin of (a)); +: cooked material; -: raw material. (b) and (c) are aligned with (a) to allow estimating the MWs on the immunoblot, as MW markers do not stain in immunoblot.
Figure 2
Figure 2
Standard curve of the sandwich ELISA for clam protein. The graph shows the response of various concentrations of clam protein, expressed as parts per million clam protein relative to unextracted sample weight.
See this image and copyright information in PMC

References

    1. Ruethers T., Taki A. C., Johnston E. B., et al. Seafood allergy: a comprehensive review of fish and shellfish allergens. Molecular Immunology. 2018;100:28–57. doi: 10.1016/j.molimm.2018.04.008. - DOI - PubMed
    1. Taylor S. L. Molluscan shellfish allergy. Advances in Food and Nutrition Research. 2008;54:139–177. doi: 10.1016/S1043-4526(07)00004-6. - DOI - PubMed
    1. Gupta R. S., Springston E. E., Warrier M. R., et al. The prevalence, severity, and distribution of childhood food allergy in the United States. Pediatrics. 2011;128(1):e9–e17. doi: 10.1542/peds.2011-0204. - DOI - PubMed
    1. Gupta R. S., Warren C. M., Smith B. M., et al. Prevalence and severity of food allergies among US adults. JAMA Network Open. 2019;2(1, article e185630) doi: 10.1001/jamanetworkopen.2018.5630. - DOI - PMC - PubMed
    1. Sicherer S. H., Munoz-Furlong A., Sampson H. A. Prevalence of seafood allergy in the United States determined by random telephone survey. Journal of Allergy and Clinical Immunology. 2004;114(1):159–165. doi: 10.1016/j.jaci.2004.04.018. - DOI - PubMed

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