Development and characterization of sandwich-type enzyme-linked aptamer assay for the detection of rongalite in food
Development and characterization of sandwich-type enzyme-linked aptamer assay for the detection of rongalite in food
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用于检测食品中雕白粉的夹心型酶联适体检测方法的开发和表征
DOI：
10.1016/j.ab.2018.09.019
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发表时间：
2018-12-15
影响因子：
2.9
通讯作者：
Han, Qinqin
中科院分区：
文献类型：
--
作者：
Jing, Le;Li, Jie;Han, Qinqin
Rongalite本质上是一种强致癌物，由于其漂白和增白的特性，被非法添加到食品加工中。本研究以荧光素酰胺（FAM）修饰的适体G 02为捕获探针，以生物素修饰的适体C 01为信号元件，建立了一种酶联适体分析方法（ELAA）。在保险粉的存在下，产生适体G 02-保险粉-适体C 01复合物，随后可以测量吸光度值。ELAA检测保险粉具有高特异性和亲和力，K-D值为19.91 +/- 1.321 nM。此外，建立了标准曲线，定量限（LOQ）为10 ng mL（-1）的保险粉。通过计算标准曲线的斜率和空白值的标准偏差，确定方法的检出限。(MDL)为0.572 ng mL（-1）。此外，在真实的食品样品中证明了ELAA的准确性。与高压液相色谱法相比，该方法可直接检测保险粉，在前处理、操作技术和成本上具有较大优势。简而言之，我们的数据表明，ELAA型可能适用于作为一种分子检测技术的保险粉。
Rongalite is an essentially strong carcinogen, which due to its properties as a bleaching and brightening, is illegally added to the food processing. In this study, a sandwich-type enzyme-linked aptamer assay (ELAA) is developed by using a rongalite-specific aptamer G02 modified fluorescein amidite (FAM) as a capture probe and aptamer C01 modified biotin as a signal element. In the presence of rongalite, the aptamer G02-rongalite-aptamer C01 complex is produced, and the absorbance value can be subsequently measured. The sandwich-type ELAA was shown to detect rongalite with high specificity and affinity, with a K-D value of 19.91 +/- 1.321 nM. In addition, the standard curve was established, with the limit of quantification (LOQ) for rongalite at 10 ng mL(-1). By calculating the slope of the standard curve and the standard deviation of the blank values, the method detection limit. (MDL) was 0.572 ng mL(-1). Additionally, the accuracy of the sandwich-type ELAA was demonstrated in real food samples. Compared with high-pressure liquid chromatography (HPLC) assay, the sandwich-type ELAA can detect rongalite directly, and it has great advantages in pre-treatment, operation technique and cost. In short, our data suggest that the sandwich-type ELAA may be applicable as a molecular detection technique for rongalite.