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利用胶体金在基于生物层干涉法的免疫传感器中对真菌毒素脱氧雪腐镰刀菌烯醇进行信号放大。

Signal amplification using colloidal gold in a biolayer interferometry-based immunosensor for the mycotoxin deoxynivalenol.

机构信息

United States Department of Agriculture, Agricultural Research Service, National Center for Agricultural Utilization Research, Peoria, IL 61604, USA.

出版信息

Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2012;29(7):1108-17. doi: 10.1080/19440049.2012.671789. Epub 2012 Apr 11.

Abstract

Deoxynivalenol (DON) is a toxin produced by certain species of Fusarium fungi that can infest wheat, barley and corn. The fungi cause diseases in crops worldwide and some of the secondary metabolites, such as DON, can adversely affect animal health and food safety. To monitor DON in wheat rapidly, a biosensor using the principle of biolayer interferometry (BLI) was developed. The signal from the sensor was substantially amplified through the use of a primary antibody-colloidal gold conjugate. The amplification was much greater in the presence of wheat matrix than in buffered solution, suggesting matrix components may have contributed to the enhancement. The improved signal provided by the amplification allowed for the development of rapid qualitative and quantitative assays. The limit of detection of the method was 0.09 mg kg(-1); the limit of quantitation was 0.35 mg kg(-1). Recovery from wheat spiked over the range from 0.2 to 5 mg kg(-1) averaged 103% (RSD = 12%). The quantitative assay compared favourably (r(2) = 0.9698) with a reference chromatographic method for 40 naturally contaminated wheats. The qualitative assay was able to classify accurately the same group of 40 samples as either above or below a 0.5 mg kg(-1) threshold. These results suggest that the BLI technique can be used to measure DON in wheat rapidly.

摘要

脱氧雪腐镰刀菌烯醇(DON)是某些镰刀菌真菌产生的一种毒素,可感染小麦、大麦和玉米。这些真菌在全球范围内引起农作物病害,其中一些次级代谢物,如 DON,会对动物健康和食品安全产生不利影响。为了快速监测小麦中的 DON,开发了一种基于生物层干涉(BLI)原理的生物传感器。通过使用一级抗体-胶体金缀合物,传感器的信号得到了极大的放大。在存在小麦基质的情况下,放大效果比在缓冲溶液中大得多,这表明基质成分可能对增强作用有贡献。放大提供的改进信号允许快速定性和定量测定的开发。该方法的检测限为 0.09 毫克/千克(-1);定量限为 0.35 毫克/千克(-1)。从 0.2 至 5 毫克/千克(-1)范围内添加的小麦的回收率平均为 103%(RSD = 12%)。定量测定与 40 种天然污染小麦的参考色谱方法相比具有良好的可比性(r(2) = 0.9698)。定性测定能够准确地将同一组 40 个样本分类为是否超过或低于 0.5 毫克/千克(-1)的阈值。这些结果表明,BLI 技术可用于快速测量小麦中的 DON。

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