Enzyme-linked immunosorbent assay for detection of zearalenone in corn, wheat, and pig feed: collaborative study.
Enzyme-linked immunosorbent assay for detection of zearalenone in corn, wheat, and pig feed: collaborative study.
复制标题
用于检测玉米、小麦和猪饲料中玉米赤霉烯酮的酶联免疫吸附测定：合作研究。
DOI：
10.1093/jaoac/77.6.1500
复制
发表时间：
1994
影响因子：
1.6
通讯作者：
Brjnton M Miller
中科院分区：
文献类型：
--
作者：
Glenn A. Bennett;Terry C. Nelsen;Brjnton M Miller
在一项国际合作研究中，23个合作者（22个实验室）对玉米、小麦和饲料中500 ng/g玉米赤霉烯酮的直接竞争性酶联免疫吸附测定（ELISA）筛选方法进行了评价。申办实验室制备了18份加标或自然污染的玉米、小麦和猪饲料样品，并将其与完整的检测试剂盒一起发送给加拿大、意大利、瑞典、荷兰和美国的参与合作者进行检测。通过在腕动振荡器上振荡3 min，用甲醇-水溶液（70 + 30）提取供试品。将一部分提取物与等体积的玉米赤霉烯酮-酶结合物混合，并将混合物与包被在微量滴定威尔斯孔上的玉米赤霉烯酮特异性单克隆抗体一起孵育。所有供试品均重复测定两次。52份空白样品中有1份（2%）报告为阳性。52份加标500 ng/g的样品中有39份（75%）报告为阳性。浓度等于或高于1000 ng/g的51份样本中有49份（96%）报告为阳性。ELISA法的总假阴性率为6.0%，假阳性率为22.7%。对于含量≥ 800 ng/g的样品，仅报告了1例（3.4%）假阴性。在分光光度法中，8名合作者根据加标提取物构建的标准曲线确定了测试样品中玉米赤霉烯酮的近似水平（每种测试商品0-3000 ng/g）。该方法给出了5.7%的假阴性和17.8%的假阳性的总体发生率。平均相对标准偏差，RSDr（重复性）和RSDR（再现性），分别为11.6%和25.1%的加标样品和11.7%和33.1%的自然污染的样品，分别。用测定的每组样品构建标准曲线。这些标准曲线的吸光度值的比较表明了测定中使用的试剂和抗体的性能。AOAC INTERNATIONAL首次采用ELISA法作为玉米、小麦和猪饲料中玉米赤霉烯酮含量≥ 800 ng/g的筛选方法。
A direct competitive enzyme-linked immunosorbent assay (ELISA) screening method for zearalenone in corn, wheat, and feed at 500 ng/g was evaluated by 23 collaborators (22 laboratories) in an international collaborative study. Eighteen samples of spiked or naturally contaminated corn, wheat, and pig feed were prepared by the sponsoring laboratory and sent for testing with complete test kits to participating collaborators in Canada, Italy, Sweden, The Netherlands, and the United States. Test samples were extracted with methanol-water solution (70 + 30) by shaking on a wrist-action shaker for 3 min. A portion of the extract was mixed with an equal volume of zearalenone-enzyme conjugate, and the mixture was incubated with zearalenone-specific monoclonal antibodies coated onto microtiter wells. All test samples were assayed in duplicate. One of 52 (2%) blanks was reported positive. Thirty-nine of the 52 (75%) samples that were spiked at 500 ng/g were reported as positive. Forty-nine of the 51 (96%) samples with concentrations at or above 1000 ng/g were reported as positive. The overall incidence of false negatives was 6.0% and the incidence of false positives was 22.7% by the ELISA method. Only one (3.4%) false negative was reported for samples containing > or = 800 ng/g. In the spectrophotometric method, 8 collaborators determined approximate levels of zearalenone in test samples from standard curves constructed from spiked extracts (0-3000 ng/g of each commodity tested). This method gave and overall incidence of false negatives of 5.7% and false positives of 17.8%. Average relative standard deviations, RSDr (repeatability) and RSDR (reproducibility), were 11.6 and 25.1% for spiked samples and 11.7 and 33.1% for naturally contaminated samples, respectively. Standard curves were constructed with each set of samples assayed. Comparison of absorbance values from these standard curves indicate the performance of reagents and antibody used in the assay. The ELISA method has been adopted first action by AOAC INTERNATIONAL as a screening method for zearalenone at > or = 800 ng/g in corn, wheat, and pig feed.