Single Laboratory Validation of a Surface Plasmon Resonance Biosensor Screening method for Paralytic Shellfish Poisoning Toxins

摘要

A research element of the European Union (EU) sixthnFramework project BioCop focused on the developmentnof a surface plasmon resonance (SPR) biosensor assaynfor the detection of paralytic shellfish poisoning (PSP)ntoxins in shellfish as an alternative to the increasinglynethically unacceptable mouse bioassay. A biosensor assaynwas developed using both a saxitoxin binding protein andnchip surface in tandem with a highly efficient simplenextraction procedure. The present report describes thensingle laboratory validation of this immunological screeningnmethod, for this complex group of toxins with differingntoxicities, according to the European Decision 2002657/EC in conjunction with IUPAC and AOAC singlenlaboratory validation guidelines. The different performancencharacteristics (detection capability CCu0001, specificityselectivity, repeatability, reproducibility, stability, andnapplicability) were determined in relation to the EUnregulatory limit of 800 μg of saxitoxin equivalents (STXneq) per kg of shellfish meat. The detection capability CCu0001nwas calculated to be 120 μg/kg. Intra-assay repeatabilitynwas found to be between 2.5 and 12.3% and interassaynreproducibility was between 6.1 and 15.2% for differentnshellfish matrices. Natural samples were also evaluatednand the resultant data displayed overall agreements of 96nand 92% with that of the existing AOAC approved methodsnof mouse bioassay (MBA) and high performance liquidnchromatography (HPLC), respectively.