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)
- Institution
- 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
A competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater, shellfish, and mammalian body fluid
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Additional Information
- Publication
- 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
- Keywords
- Algal toxins--Florida, Dinoflagellates--Florida, Enzyme-linked immunosorbent assay, Neurotoxic agents--Florida, Ptychodiscus brevis--Florida, Seafood--Contamination--Florida, Shellfish as food--Contamination--Florida
- Subjects
- Neurotoxic agents -- Florida
- Algal toxins -- Florida
- Dinoflagellates -- Florida
- Ptychodiscus brevis -- Florida
- Enzyme-linked immunosorbent assay
- Seafood--Contamination--Florida
- Shellfish as food--Contamination--Florida