Polakowski Sergiusz, Davis Alan H, Albert Andre L, Gow Brendan
EnviroLogix Inc., 500 Riverside Industrial Pkwy, Portland, ME 04103, USA.
J AOAC Int. 2015 Jan-Feb;98(1):85-93. doi: 10.5740/jaoacint.14-152.
Lateral flow technology and a reader-based system are used for quantitative determination of deoxynivalenol (DON), also known as vomitoxin, residues in cereal grain commodities by the QuickTox Kit for QuickScan DON (Vomitoxin). The assay has been modified, and a study was conducted in support of a Performance Tested MethodSM (PTM) Modification. The modified assay employs identical biologic reagents as used previously (PTM No. 121202). Compared to the PTM certified product, the new assay uses modified device architecture. Multiple kits and catalog numbers were required in the original kit reflecting the necessity for matrix specific calibration curves affixed to assay strips. A single calibration curve and kit are utilized in the new product; extraction volumes used in sample preparation are varied to accommodate multiple sample types. Extracts are clarified by filtration or settling depending on the sample type. Filtration was used for matrixes examined in these studies. With the original product, the extract was mixed 1:1 with DB1 buffer followed by the addition of the strip which was developed for 10 min. The new product dilutes extracts five-fold offline in DB6 buffer; an aliquot of the dilution is moved to a reaction vial followed by strip development time for 3 min. The new assay performance was evaluated for linearity, robustness, selectivity (inclusivity), lot-to-lot consistency, and both internal and third party matrix studies. All DON positive samples yielded results within previously defined acceptable ranges with dose-dependent correlation values of R2 greater than 0.97 in linearity and internal and external matrix studies. Inclusivity data indicated detection of DON along with acetyl derivatives, glucoside-conjugate, and Nivalenol. Robustness studies showed within range results upon co-variation of multiple user interface parameters, and lot-to-lot consistency challenges demonstrated acceptable results across five manufactured lots.
横向流动技术和基于读数器的系统被用于通过QuickTox试剂盒（用于QuickScan DON（呕吐毒素））对谷物商品中的脱氧雪腐镰刀菌烯醇（DON，又称呕吐毒素）残留进行定量测定。该检测方法已进行了修改，并开展了一项研究以支持性能测试方法SM（PTM）修改。修改后的检测方法使用与之前相同的生物试剂（PTM编号121202）。与经过PTM认证的产品相比，新的检测方法采用了改进的设备架构。原始试剂盒需要多个试剂盒和产品目录编号，这反映了为检测条粘贴特定基质校准曲线的必要性。新产品使用单一校准曲线和试剂盒；样品制备中使用的提取体积有所不同，以适应多种样品类型。提取物根据样品类型通过过滤或沉降进行澄清。在这些研究中，对所检测的基质使用过滤方法。对于原始产品，提取物与DB1缓冲液按1:1混合，然后加入检测条，显色10分钟。新产品在DB6缓冲液中离线将提取物稀释五倍；取一份稀释液转移至反应瓶中，然后检测条显色3分钟。对新检测方法的线性、稳健性、选择性（包容性）、批次间一致性以及内部和第三方基质研究进行了评估。在内部和外部基质研究的线性测试中，所有DON阳性样品的结果均在先前定义的可接受范围内，剂量依赖性相关值R2大于0.97。包容性数据表明能够检测到DON及其乙酰衍生物、葡萄糖苷共轭物和雪腐镰刀菌烯醇。稳健性研究表明，在多个用户界面参数共同变化时结果在范围内，批次间一致性测试表明在五个生产批次中均取得了可接受的结果。
