title: Comparison of a protein phosphatase inhibition assay, HPLC assay and enzyme-linked immunosorbent assay with the mouse bioassay for the detection of diarrhetic shellfish poisoning toxins in European shellfish.
abstract: Consumption of shellfish contaminated with algal toxins produced by marine dinoflagellates can lead to diarrhetic shellfish poisoning (DSP). UK legislation necessitates toxin detection by mouse bioassay but this method is non-specific and lacks sensitivity. As an alternative method, an HPLC technique has been optimized, with detection limits of 0.26 micrograms of toxin/g of shellfish hepatopancreas for both Okadaic Acid (OA) and Dinophysistoxin-1 (DTX-1). A colorimetric protein phosphatase inhibition (PPI) assay has also been optimized. This assay detects inhibition of protein phosphatase 1 (PPI gamma) by OA and DTX-1 with detection limits of 1.5 ng of total toxin/g of hepatopancreas. Contaminated shellfish from several European sources, the UK monitoring programmes and mussels associated with an outbreak of DSP poisoning in the UK, have been analyzed and assessed using the two alternative methods and a commercially available enzyme-linked immunosorbent assay (ELISA) kit. The results indicate that both the HPLC and PPI assays correlate well with each other and with the UK standard mouse bioassay. In contrast, and not withstanding its advantages of rapidity and ease, the ELISA kit did not accurately and consistently detect low toxin concentrations, although it may be useful as a screening tool.
authors.affiliation: Food Hygiene Laboratory, PHLS Central Public Health Laboratory, Colindale, London, UK.
authors.forename: P E
authors.initials: PE
authors.lastname: Núñez
authors.forename: A C
authors.initials: AC
authors.lastname: Scoging
mesh_terms: D000818:Animals; D001681:Biological Assay; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003124:Colorimetry; D003967:Diarrhea / Q000139:chemically induced* / Q000453:epidemiology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005060:Europe / Q000453:epidemiology; D005517:Foodborne Diseases / Q000453:epidemiology / Q000209:etiology; D006801:Humans; D008099:Liver / Q000737:chemistry; D008387:Marine Toxins / Q000009:adverse effects* / Q000032:analysis*; D051379:Mice; D008807:Mice, Inbred BALB C; D019319:Okadaic Acid / Q000032:analysis / Q000031:analogs & derivatives; D010179:Pancreas / Q000737:chemistry; D010749:Phosphoprotein Phosphatases / Q000032:analysis*; D054645:Protein Phosphatase 1; D011714:Pyrans / Q000032:analysis; D012680:Sensitivity and Specificity; D012758:Shellfish / Q000032:analysis*; D006113:United Kingdom / Q000453:epidemiology
chemical_list: D008387:Marine Toxins; D011714:Pyrans; D019319:Okadaic Acid; C051904:dinophysistoxin 1; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1