Using of Tyramine Signal Amplification to Improve the Sensitivity of ELISA for Aflatoxin B1 in Edible Oil Samples
Using of Tyramine Signal Amplification to Improve the Sensitivity of ELISA for Aflatoxin B1 in Edible Oil Samples
复制标题
利用酪胺信号放大提高 ELISA 对食用油样品中黄曲霉毒素 B-1 的灵敏度
DOI：
10.1007/s12161-018-1235-9
复制
发表时间：
2018-09-01
影响因子：
2.9
通讯作者：
Du, Daolin
中科院分区：
文献类型：
--
作者：
Zhang, Yuanyuan;Li, Ming;Du, Daolin
酪胺信号放大（TSA）是一种优良的信号放大策略，具有提高分析物分析灵敏度的潜力。本研究将TSA系统与生物素-链亲和素（BSA）系统偶联后，酶联免疫吸附试验（ELISA）的灵敏度显著提高。为此，建立了一种基于TSA的灵敏TSA- elisa检测食用油中黄曲霉毒素B-1 （AFB(1)）的方法。在最佳条件下，TSA-ELISA对AFB(1)的检出限（LOD, IC10）和半最大抑制浓度（IC50）分别为0.004和0.039 ng/mL。开发的AFB(1) TSA-ELISA与ELISA相比LOD值提高了11倍，IC50值提高了6倍。TSA-ELISA与类似物的交叉反应性可忽略不计（< 3.48%），具有较高的特异性。AFB(1)在食用油样品中的加标回收率为81.4 ~ 118.8%，相对标准偏差为3.8 ~ 9.0%。此外，TSA-ELISA检测结果与hplc -荧光检测结果具有良好的相关性。TSA-ELISA是一种灵敏、廉价、高通量、可替代的食用油中AFB(1)检测方法。本研究为提高ELISA的灵敏度提供了一种简单实用的方法，具有重要的推广价值和应用前景。
Tyramine signal amplification (TSA), the excellent signal amplification strategy, has the potential to improve the sensitivities of analytes analysis. In this work, the sensitivity of enzyme-linked immunosorbent assays (ELISA) has significantly improved by coupling TSA system and further biotin-streptavidin (BSA) system. Thus, a sensitive TSA-ELISA based on TSA was developed for detecting aflatoxin B-1 (AFB(1)) in edible oil samples. Under optimal conditions, the limit of detection (LOD, IC10) and the half-maximal inhibition concentration (IC50) of the TSA-ELISA were 0.004 and 0.039 ng/mL for AFB(1), respectively. The developed TSA-ELISA for AFB(1) has an 11-fold improved LOD value and 6-fold improved IC50 value when compared with ELISA. The cross-reactivities of the TSA-ELISA with its analogues were negligible (< 3.48%), which indicated high specificity. The spiked recoveries were 81.4 to 118.8% with relative standard deviations (RSDs) of 3.8 to 9.0% for AFB(1) in edible oil samples. Furthermore, the results of TSA-ELISA correlated well with those obtained by HPLC-fluorescence detector. The proposed TSA-ELISA was a satisfactory tool for sensitive, inexpensive, high-throughput, and alternative detection of AFB(1) in edible oil samples. This study could provide the strategy for improving the sensitivity of ELISA with simple and practical approach, which has significant popularizing value and application prospect.