Single Laboratory Validation of a Surface Plasmon Resonance Biosensor Screening method for Paralytic Shellfish Poisoning Toxins
Single Laboratory Validation of a Surface Plasmon Resonance Biosensor Screening method for Paralytic Shellfish Poisoning Toxins
复制标题
DOI：
10.1021/ac1000338
复制
发表时间：
2010-04-01
影响因子：
7.4
通讯作者：
Elliott, Christopher T.
中科院分区：
文献类型：
--
作者：
Campbell, Katrina;Haughey, Simon A.;Elliott, Christopher T.
欧洲联盟（欧盟）第六个框架项目BioCop的一项研究内容侧重于开发一种表面等离子体共振（SPR）生物传感器测定法，用于检测贝类中的麻痹性贝类中毒毒素，作为道德上越来越不可接受的小鼠生物测定法的替代方法。一个生物传感器检测开发使用石房蛤毒素结合蛋白和芯片表面串联与一个高效的简单的提取程序。本报告描述了根据欧洲决定2002/657/EC以及IUPAC和AOAC单一实验室验证指南，针对这一具有不同毒性的复杂毒素组，对这种免疫学筛选方法进行的单一实验室验证。不同的性能特征（检测能力CC β，特异性/选择性，重复性，再现性，稳定性和适用性）进行了测定与欧盟的监管限制800微克的蛤蚌毒素当量（STX当量）每公斤贝类肉。计算出检测能力CC β为120 μ g/kg。不同贝类基质的批内重复性在2.5 - 12.3%之间，批间重复性在6.1 - 15.2%之间。天然样品也进行了评估，所得数据显示，与现有的AOAC批准的小鼠生物测定（MBA）和高效液相色谱法（HPLC）的方法，分别为96%和92%的总体协议。
A research element of the European Union (EU) sixth Framework project BioCop focused on the development of a surface plasmon resonance (SPR) biosensor assay for the detection of paralytic shellfish poisoning (PSP) toxins in shellfish as an alternative to the increasingly ethically unacceptable mouse bioassay. A biosensor assay was developed using both a saxitoxin binding protein and chip surface in tandem with a highly efficient simple extraction procedure. The present report describes the single laboratory validation of this immunological screening method, for this complex group of toxins with differing toxicities, according to the European Decision 2002/657/EC in conjunction with IUPAC and AOAC single laboratory validation guidelines. The different performance characteristics (detection capability CC beta, specificity/selectivity, repeatability, reproducibility, stability, and applicability) were determined in relation to the EU regulatory limit of 800 mu g of saxitoxin equivalents (STX eq) per kg of shellfish meat. The detection capability CC beta was calculated to be 120 mu g/kg. Intra-assay repeatability was found to be between 2.5 and 12.3% and interassay reproducibility was between 6.1 and 15.2% for different shellfish matrices. Natural samples were also evaluated and the resultant data displayed overall agreements of 96 and 92% with that of the existing AOAC approved methods of mouse bioassay (MBA) and high performance liquid chromatography (HPLC), respectively.