Lexmaulová Hana, Gabrovská Dana, Rysová Jana, Stumr Frantisek, Netusilová Katerina, Blazková Martina, Bulawová Hana, Brychta Josef, Subrtová Zdenka, Pavelka Jiri, Iametti Stefania, Del Barco Jorge Antonio Guisantes, Quesada Jorge Martinez, Pardo Esther Sunen, Resa Idoia Postigo, Takkinen Kristiina, Laukkanen Marja-Leena, Piknová Lubica, Langerholc Tomaz, Cencic Avrelija, Barsová Sona, Cuhra Petr, Plicka Jan
ELISA Development, Ltd, 412 01 Velk Zernoseky 186, Czech Republic.
J AOAC Int. 2013 Sep-Oct;96(5):1041-7. doi: 10.5740/jaoacint.12-474.
A collaborative study in 10 laboratories was performed to validate an ELISA method developed for the quantitative determination of peanut protein in foods. The ELISA kit used for this study is based on rabbit polyclonal antibody. This kit does not produce any false-positive results or cross-reactivity with a broad range of peanut-free food matrixes. All participants obtained the peanut ELISA kit with standard operational procedures, a list of samples, the samples, and a protocol for recording test results. The study included 15 food samples. Three food matrix samples of zero peanut content showed peanut protein content lower than the first standard (0.10 mg/kg). Three samples with peanut declared as an ingredient revealed peanut protein content outside the calibration curve (absorbance was above the highest standard) in all laboratories, and three samples had the peanut content reported either above the highest standard or within the calibration curve, depending on the laboratory. Six samples with peanut declared as an ingredient gave the peanut protein content within the calibration curve. Only these six samples, together with a positive control sample (CS2), were used for statistical evaluation. The statistical tests (Cochran, Grubbs, and Mandel) and analysis of variance were used for the evaluation of the collaborative study results. Repeatability and reproducibility limits, as well as an LOQ (LOQcollaborative 0.22 mg peanut proteins/kg) and an LOD (LODcollaborative 0.07 mg peanut proteinslkg) for the kit were calculated.
在10个实验室开展了一项合作研究，以验证一种为定量测定食品中花生蛋白而开发的酶联免疫吸附测定（ELISA）方法。本研究使用的ELISA试剂盒基于兔多克隆抗体。该试剂盒不会产生任何假阳性结果，也不会与多种不含花生的食品基质发生交叉反应。所有参与者均获得了带有标准操作程序、样品清单、样品以及测试结果记录方案的花生ELISA试剂盒。该研究包括15个食品样品。三个花生含量为零的食品基质样品显示花生蛋白含量低于第一个标准（0.10毫克/千克）。三个标明含有花生成分的样品在所有实验室中均显示花生蛋白含量在校准曲线之外（吸光度高于最高标准），还有三个样品的花生含量报告高于最高标准或在校准曲线范围内，具体取决于实验室。六个标明含有花生成分的样品的花生蛋白含量在校准曲线内。仅将这六个样品以及一个阳性对照样品（CS2）用于统计评估。使用统计检验（ Cochr an检验、Grubbs检验和Mandel检验）和方差分析来评估合作研究结果。计算了该试剂盒的重复性和再现性限，以及定量限（合作定量限为0.22毫克花生蛋白/千克）和检测限（合作检测限为0.07毫克花生蛋白/千克）。
