The development of a diagnostic tool for ciguatera fish poisoning in human serum.
AuthorGamboa Pulido, Pedro Miguel.
Committee ChairPark, Douglas L.
MetadataShow full item record
PublisherThe University of Arizona.
RightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
AbstractThe development of a clinical diagnostic scheme for ciguatera fish poisoning has been accomplished through the utilization of three assays, specifically a competitive enzyme-linked immunosorbent assay (ELISA) for quantification, and two procedures for separation of toxic potentials, i.e., a liquid-liquid partition method and an immunoaffinity column system. For the ELISA assay, quality control parameters were evaluated including the coating binding efficiency and the binding capability of the conjugate (okadaic acid-bovine serum albumin). A 100% recovery was obtained and reproducibility values were under 30% coefficient of variation (CV) for concentrations from 0.0001 to 100 ng okadaic acid/mL serum or g fish flesh. Additionally, the usefulness of the method was evaluated by determining sensitivity and specificity parameters (66.7 and 95%, respectively) from where the positive and negative predictive values of the test as a diagnostic instrument were assessed. A rapid extraction procedure utilizing a mixture of methanol:chloroform:water was adapted for extraction and isolation of toxins associated with ciguatera fish poisoning. The method has been optimized for the analytical evaluation of potentially ciguatoxic fish samples and human serum. Applications of the extraction procedure include the clinical diagnosis of ciguatera illness and, if leftover fish portions responsible for the intoxication event are available, analysis of the seafood. It can also be used as a tool for monitoring seafood products for food safety. Recovery values of the extraction method with and without an intermediary defatting step for spiked fish samples were of 105 and 86%, respectively. By attaching anti-okadaic acid monoclonal antibody onto an inert matrix (AH-Sepharose®), an immunoaffinity column specific to ciguatera-related toxins has been developed, also for serum analysis, but with the potential application in other body fluids. The binding efficiency was 82% and reproducibility values for spiked test portions at concentrations as low as 250 pg were below 10%.
Degree ProgramNutritional Sciences