Detection of domoic acid in rat serum and brain by direct competitive enzyme-linked immunosorbent assay (cELISA)

Abstract

In 1987 a large-scale incident of human poisoning in Canada was traced to commercial mussels contaminated with domoic acid (DOM). Since then, routine screening of shellfish domoic acid content has been carried out using a variety of assays, with liquid chromatography using ultraviolet absorbance detection (LC-UV) or mass spectrometric detection (LC-MS) being the currently accepted standard methodologies. Recently, a highly specific competitive enzyme-linked immunosorbent assay (cELISA) has been developed for the detection and analysis of DOM in commercial shellfish, but its accuracy relative to LC methods has not been independently verified in mammalian tissues. In this study we demonstrate that measurement of rat serum DOM concentration by cELISA gives a good correlation (r2 = 0.993) across a broad range of concentrations when compared to LC-MS analysis, with only a small (15%) overestimation of sample DOM content. In addition, we have developed an extraction method for analysis of DOM in rat brain by cELISA which yields complete recovery across a range of sample dilutions.

Ischemic Cardiomyopathy Following Seizure Induction by Domoic Acid

“…These findings are consistent with previous studies from our group, which established that intraperitoneal delivery of 2 mg/kg DOM resulted in significantly elevated levels of DOM in the brain. 44 The present study has also shown that, although cardiac levels of DOM were increased within 1 hour of intraperitoneal dosing (2 mg/kg) with the excitotoxin, localized intrahippocampal infusions with 100 pmol DOM failed to produce a detectable level of DOM in the circulation. This is not surprising, because, even though the integrity of the blood-brain barrier may be compromised during seizure activity, resulting in systemic diffusion, the original 100 pmol dose applied would have been greatly diluted.…”

“…The concentration of DOM within the diluted cardiac homogenate (100-fold) and serum (1000-fold) samples was quantified using an ASP direct cELISA kit, as described previously. 31,44 Cardiac Hemodynamics Study groups were sacrificed at the specified time points. Both DOM-and vehicle-treated rats were decapitated without anesthesia and the hearts were rapidly excised into cold (4°C), filtered Krebs-Heinsleit buffer containing (in mmol/L), 118 NaCl, 4.7 KCl, 1.2 MgSO 4 , 1.23 KH 2 PO 4 , 24 NaHCO 3 , 11.1 glucose, and 1.2 CaCl 2 (pH 7.4).…”

Development, Comparison, and Validation Using ELISAs for the Determination of Domoic Acid in California Sea Lion Body Fluids

“…It has been validated for the analysis of DA in shellfish tissues and in dissolved and particulate phytoplankton samples (35,36). The BS ELISA is a polyclonal antibody-based assay developed by Biosense Laboratories; it has been validated by both single and interlaboratory studies for the analysis of DA in shellfish tissues (37,38) and for the analysis of DA concentrations present in rat serum and brain samples (39). An Agilent (Santa Clara, CA) 6130 LC/MS system operated in positive electrospray ionization mode with an Agilent Zorbax Rapid Resolution column and selected ion monitoring of DA (312 amu) was used for LC/MS analysis generally following the method of Wang et al (40).…”

A low cost novel sensing system for detection of dangerous marine biotoxins in seafood

“…Research of using ELISA to determine DA has been reported in [7,[27][28][29][30]. ELISA method normally can be used to detect only one particular toxin.…”