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Application of an in-vitro tissue culture assay for the determination of paralytic shellfish poison (Saxitoxin family) and comparison to the HPLC assay and the standard mouse bioassay

Thorne, T.J., Cassell, H.S. and Jones, J.B.ORCID: 0000-0002-0773-2007 (1999) Application of an in-vitro tissue culture assay for the determination of paralytic shellfish poison (Saxitoxin family) and comparison to the HPLC assay and the standard mouse bioassay. Australian Government. Fisheries Research and Development Corporation

Abstract

Paralytic shellfish poisoning (PSP) is a condition that can occur when humans consumer shellfish that have accumulated the causative algal biotoxin, saxitoxin. Saxitoxin, and a group of 18 or more similar analogues, are potent neurotoxins produced by a number of dinoflagellates that cause algal blooms or red tides (Hallegraeff 1993). PSP may result in human illness and in some cases, death. As the toxin is unable to be detected outside of the laboratory it poses a major health risk.

Laboratory detection of saxitoxin can be performed by a number of methods including mouse assay, high performance liquid chromatography (HPLC), radioimmunoassay or enzyme linked immunoassay (ELISA). As there are distinct disadvantages with all these techniques, initial investigations into an in-vitro tissue culture bioassay utilizing a mouse neuroblastoma cell line has been initiated.

The purpose of this study was to investigate the possible application of the tissue culture assay in Western Australia using techniques outlined in the scientific literature from overseas. Once the assay was established in the laboratory, saxitoxin levels in a selection of ten shellfish extracts were analyzed by the tissue culture assay and compared to results obtained by way of mouse bioassay and HPLC.

Results indicate that it would be possible to establish the cell bioassay technique in any tissue culture laboratory with the appropriate equipment and personnel skilled in tissue culture techniques. The investigation revealed that although it was possible to establish the assay, the results of the analysis of the shellfish extracts were inconsistent and lacked sensitivity, although results appeared to more consistent in shellfish extracts with lower level of toxin. In order to achieve repeatable results, further investigation must be undertaken to modify the Neuro-2a cell line to ensure that the number of sodium channel receptors on the cells is consistent and of a sufficiently high number.

Item Type: Report
Series Name: Final Report. FRDC Project No. 96/323
Publisher: Australian Government. Fisheries Research and Development Corporation
URI: http://researchrepository.murdoch.edu.au/id/eprint/43206
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