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Ultrasensitive and quantitative gold nanoparticle-based immunochromatographic assay for detection of ochratoxin A in agro-products

机译:基于金纳米颗粒的超灵敏定量免疫色谱分析方法,用于检测农产品中的曲霉毒素A

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摘要

In most cases of mycotoxin detection, quantitation is critical while immunochromatographic strip tests are qualitative in nature. Moreover, the sensitivity of this technique is questioned. In order to overcome these limitations, an ultrasensitive and quantitative immunochromatographic assay (ICA) for rapid and sensitive quantitation of ochratoxin A (OTA) was developed. The assay was based on a competitive format and its sensitivity was improved by using a sensitive and selective OTA monoclonal antibody (OTA-mAb). The visible ICA results were obtained within 15 min, and in addition to visual examination, they were read by the rapid color intensity portable strip reader. The visual and computational detection limits (vLOD and cLOD, respectively) for ochratoxin A were 0.2 and 0.25 ng mL(-1), respectively. These values were lower than those reported by previous studies in a range 5-2500 folds. For validation, contaminated samples including wheat, maize, rice and soybean were assayed by ICA and a standard high performance liquid chromatography (HPLC). The results were in good agreement for both ICA and HPLC methods. The average recoveries of the HPLC were in the range 72-120% while the ICA values were from 76 to 104%, confirming the accuracy and sensitivity of this method. (C) 2014 Elsevier B.V. All rights reserved.
机译:在真菌毒素检测的大多数情况下,定量分析至关重要,而免疫层析试纸检测本质上是定性的。此外,该技术的敏感性受到质疑。为了克服这些限制,开发了一种用于sensitive曲霉毒素A(OTA)快速灵敏定量的超灵敏定量免疫色谱分析(ICA)。该测定法基于竞争性格式,并通过使用敏感的选择性OTA单克隆抗体(OTA-mAb)提高了其敏感性。可见的ICA结果在15分钟内获得,除目测外,还通过快速色强度便携式条带阅读器读取。曲霉毒素A的视觉和计算检出限(分别为vLOD和cLOD)分别为0.2和0.25 ng mL(-1)。这些值比以前的研究报告的值低5-2500倍。为了进行验证,使用ICA和标准高效液相色谱(HPLC)对包括小麦,玉米,大米和大豆在内的受污染样品进行了分析。 ICA和HPLC方法的结果均吻合良好。 HPLC的平均回收率在72-120%的范围内,而ICA值则在76%至104%的范围内,证实了该方法的准确性和灵敏度。 (C)2014 Elsevier B.V.保留所有权利。

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