Receptor binding assay for paralytic shellfish poisoning toxins: optimization and interlaboratory comparison.
A receptor binding assay (RBA) for detection of paralytic shellfish poisoning (PSP) toxins was formatted for use in a high throughput detection system using microplate scintillation counting. The RBA technology was transferred from the National Ocean Service, which uses a Wallac TriLux 1450 MicroBeta microplate scintillation counter, to the California Department of Health Services, which uses a Packard TopCount scintillation counter. Due to differences in the detector arrangement between these 2 counters, markedly different counting efficiencies were exhibited, requiring optimization of the RBA protocol for the TopCount instrument. Precision, accuracy, and sensitivity [limit of detection = 0.2 microg saxitoxin (STX) equiv/100 g shellfish tissue] of the modified protocol were equivalent to those of the original protocol. The RBA robustness and adaptability were demonstrated by an interlaboratory study, in which STX concentrations in shellfish generated by the TopCount were consistent with MicroBeta-derived values. Comparison of STX reference standards obtained from the U.S. Food and Drug Administration and the National Research Council, Canada, showed no observable differences. This study confirms the RBA's value as a rapid, high throughput screen prior to testing by the conventional mouse bioassay (MBA) and its suitability for providing an early warning of increasing PSP toxicity when toxin levels are below the MBA limit of detection.
D029845:Amphibian Proteins; D000818:Animals; D001921:Brain / Q000648:ultrastructure; D002352:Carrier Proteins / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D007753:Laboratories; D008297:Male; D011786:Quality Control; D051381:Rats; D012015:Reference Standards; D012530:Saxitoxin / Q000032:analysis* / Q000378:metabolism; D012680:Sensitivity and Specificity; D012758:Shellfish / Q000032:analysis*; D015222:Sodium Channels / Q000378:metabolism*; D014316:Tritium
D029845:Amphibian Proteins; D002352:Carrier Proteins; D015222:Sodium Channels; C027525:saxitoxin-binding protein, Rana catesbeiana; D014316:Tritium; D012530:Saxitoxin