酶联适体分析法检测葡萄酒中的赭曲霉素A

doi:10.3969/j.issn.1673-1689.2015.04.010

首页 > 过刊浏览>2015年第34卷第4期 >396-401. DOI:10.3969/j.issn.1673-1689.2015.04.010

酶联适体分析法检测葡萄酒中的赭曲霉素A
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                        10.3969/j.issn.1673-1689.2015.04.010
                    
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Enzyme-Linked Aptamer Assay for Detection of Ochratoxin A in Wine

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建立了竞争取代酶联适体分析方法,检测葡萄酒中的赭曲霉素A (OTA)。核酸适体和OTA特异性结合导致与核酸适体杂交的短链 DNA 解链,解离的 DNA 作为捕获元素,进一步特异性结合辣根过氧化物酶 (HRP),HRP催化四甲基二苯胺 (TMB)底物显色,测定A450 nm与浓度的线性关系,确定OTA的检出限。考察了DNA包被浓度、杂交温度和封闭液等因素对检测的影响。结果表明,在优化的条件下,所建立的竞争取代酶联适体分析法对OTA检测有高灵敏度,检测限 0.88 μg/L,线性范围 1～100 μg/L在葡萄酒中添加时,加标回收率为 92.03 %～106.6 %,相对标准偏差RSD(n=5)小于 2.1 %,此方法可用于葡萄酒中OTA的快速测定。

Abstract:

ompetitive enzyme-linked aptamer assay for Ochratoxin A (OTA) detection in wine was investigated in this study. The binding of aptamer to the target OTA resulted in the dissociation of the DNA short chain from the aptamer, which was further captured by horseradish peroxidase (HRP). HRP catalyzed 3,3',5,5'.-Tetramethylbenzidine (TMB) substrate and finally leaded to a characteristic change that was detectable by the absorption of ultraviolet at 450 nm. The detection limit of OTA was determined by the linear relationship between OTA concentration and the absorbance. The factors influenced the effect of detection were investigated, including DNA concentration, temperature of hybridization, and blocking buffer solution. This work described the high performance of the competitive enzyme-linked aptamer assay (ELAA) for the determination of OTA. Under the optimal condition, the limit of detection attained was 0.88 μg/L and the linearity was in the range 1 to 100 μg/L. For the OTA detection in wine, the recovery rate was between 92.03% and 106.6%, and the relative standard deviation (RSD, n=5) was within 2.1%. This study validates the competitive ELAA is a useful tool for a rapid detection of OTA in wine.

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赵秋伶,史素青.酶联适体分析法检测葡萄酒中的赭曲霉素A[J].食品与生物技术学报,2015,34(4):396-401.

ZHAO Qiuling, SHI Suqing. Enzyme-Linked Aptamer Assay for Detection of Ochratoxin A in Wine[J]. Journal of Food Science and Biotechnology,2015,34(4):396-401.

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在线发布日期: 2015-08-09
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