A competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater, shellfish, and mammalian body fluid

UNCW Author/Contributor (non-UNCW co-authors, if there are any, appear on document)
Daniel G Baden, Director (Creator)
Andrea J. Bourdelais, Research Associate Professor (Creator)
Jerome Naar, Research Associate Professor (Creator)
Carmelo R. Tomas, Associate Professor (Creator)
The University of North Carolina Wilmington (UNCW )
Web Site: http://library.uncw.edu/

Abstract: We developed a competitive enzyme-linked immunosorbent assay (ELISA) to analyze brevetoxins, using goat anti-brevetoxin antibodies obtained after immunization with keyhole limpet hemocyanin- brevetoxin conjugates, in combination with a three-step signal amplification process. The procedure, which used secondary biotinylated antibodies, streptavidine-horseradish peroxidase conjugate, and chromogenic enzyme substrate, was useful in reducing nonspecific background signals commonly observed with complex matrices. This competitive ELISA detected brevetoxins in seawater, shellfish extract and homogenate, and mammalian body fluid such as urine and serum without pretreatment, dilution, or purification. We investigated the application of this technique for shellfish monitoring by spiking shellfish meat with brevetoxins and by analyzing oysters from two commercial shellfish beds in Florida that were exposed to a bloom of Karenia brevis (formerly Gymnodinium breve). We performed brevetoxin analysis of shellfish extracts and homogenates by ELISA and compared it with the mouse bioassay and receptor binding assay. The detection limit for brevetoxins in spiked oysters was 2.5 µg/100 g shellfish meat. This assay appears to be a useful tool for neurotoxic shellfish poisoning monitoring in shellfish and seawater, and for mammalian exposure diagnostics, and significantly reduces the time required for analyses. Key words: antibody, brevetoxin, detection, ELISA, immunoassay, neurotoxic shellfish poisoning, PbTx, seawater, serum, shellfish, urine. Environ Health Perspect 110:179–185 (2002). [Online 17 January 2002] http://ehpnet1.niehs.nih.gov/docs/2002/110p179-185naar/abstract.html

Additional Information

Naar, J., Bourdelais, A., Tomas, C., Kubanek, J., Whitney, P. L., Flewelling, L., et al. (2002). A competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater,shellfish, and mammalian body fluid. Environmenal Health Perspectives, 110, 179-185. Retrieved from http://www.ehponline.org/docs/2002/110p179-185naar/abstract.html [Reproduced with permission from Environmental Health Perspectives.]
Language: English
Date: 2009
Algal toxins--Florida, Dinoflagellates--Florida, Enzyme-linked immunosorbent assay, Neurotoxic agents--Florida, Ptychodiscus brevis--Florida, Seafood--Contamination--Florida, Shellfish as food--Contamination--Florida
Neurotoxic agents -- Florida
Algal toxins -- Florida
Dinoflagellates -- Florida
Ptychodiscus brevis -- Florida
Enzyme-linked immunosorbent assay
Shellfish as food--Contamination--Florida

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