Quantitative Method for Ciguatera Screening

As worldwide demand for fish products increases, ciguatera fish poisoning is becoming an expanding circumglobal problem. The need for seafood safety requires a rapid, quantitative method to detect ciguatera toxins in fish. Preliminary studies of a membrane immunobead assay (MIA) using fluorescent particles (FMIA) have shown it to be a rapid method to quantify ciguatoxin (CTX). Fluorescent spectral data, quantified with a spectrometer, demonstrated that the fluorescent immunobeads reacted with fish extracts containing CTX in a concentration-dependent manner. Scanning electron microscopy (SEM) photo analyses confirmed these results, supporting the feasibility of the FMIA method to detect CTX. In addition, two prototype fluorometers, one for laypersons and the other for laboratory or industrial use, have been designed and fabricated specifically to detect the fluorescent signal from the FMIA samples. Initial studies show that this fluorometer could distinguish between toxic and non-toxic FMIA samples. This project will focus on refining the FMIA procedure; quantifying samples prepared with pure CTXs, toxic fish extracts, and from reef fish associated with ciguatera; verifying toxicity of these samples with the MIA and neuroblastoma cell assay; improving the prototype fluorometers; and calibrating the fluorometers to accurately quantify FMIA samples.