Stumr Frantisek, Gabrovská Dana, Rysová Jana, Hanák Petr, Plicka Jan, Tomková Kveta, Cuhra Petr, Kubík Martin, Barsová Sona, Karsulínoví Lenka, Bulawová Hana, Brychta Josef
SEDIUM RD, s.r.o., Zeleznicního pluku 1361, 530 02 Pardubice, Czech Republic.
J AOAC Int. 2009 Sep-Oct;92(5):1519-25.
An interlaboratory study was performed in six laboratories to prove the validation of the ELISA method developed for quantitative determination of beta-lactoglobulin (BLG) in foods. The ELISA kit used for this study is based on rabbit polyclonal antibody. In-house validation of the kit did not produce false-positive results or cross-reactivity in a broad range of food matrixes containing no milk proteins. All participants obtained the BLG kit with a standard operational procedure, the list of the samples, samples, and a protocol for recording test results. The study included 14 food samples (extruded breakfast cereals, bread, two soy desserts, butter, chicken ham, chicken meat, wheat flour, long grain rice, jelly, two whey drinks, crackers, and bitter chocolate) and six spiked samples (two rice, two wheat flour, and two chicken meat). Nine samples of food matrixes containing no milk proteins showed BLG content lower than the first standard (0.15 mg/kg). Two samples of food matrixes with no milk proteins revealed BLG content higher than standard 3 (1.5 mg/100 g) and standard 4 (5.0 mg/100 g). Three food samples containing milk were tested as positive, and all spiked samples were evaluated as positive. The statistical tests (Cochran, Dixon, and Mandel) and analysis of variance were used to evaluate the interlaboratory study results. Repeatability and reproducibility limits, as well as LOQ (0.22 mg BLG/kg) and LOD (0.07 mg BLG/kg), for the kit were calculated.
在六个实验室进行了一项实验室间研究，以验证所开发的用于定量测定食品中β-乳球蛋白（BLG）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。该试剂盒的内部验证在不含乳蛋白的多种食品基质中未产生假阳性结果或交叉反应。所有参与者都按照标准操作程序获得了BLG试剂盒、样品清单、样品以及记录测试结果的方案。该研究包括14种食品样品（挤压早餐谷物、面包、两种大豆甜点、黄油、鸡肉火腿、鸡肉、小麦粉、长粒米、果冻、两种乳清饮料、饼干和苦巧克力）和6种加标样品（两种大米、两种小麦粉和两种鸡肉）。9种不含乳蛋白的食品基质样品的BLG含量低于第一个标准（0.15毫克/千克）。两种不含乳蛋白的食品基质样品的BLG含量高于标准3（1.5毫克/100克）和标准4（5.0毫克/100克）。三种含乳食品样品检测为阳性，所有加标样品评估为阳性。使用统计检验（ Cochr an、Dixon和Mandel检验）和方差分析来评估实验室间研究结果。计算了该试剂盒的重复性和再现性限，以及定量限（0.22毫克BLG/千克）和检测限（0.07毫克BLG/千克）。
