Immunochromatographic assay for ultrasensitive detection of aflatoxin B₁ in maize by highly luminescent quantum dot beads.
Immunochromatographic assay for ultrasensitive detection of aflatoxin B₁ in maize by highly luminescent quantum dot beads.
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DOI：
10.1021/am503517s
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发表时间：
2014-08-27
影响因子：
9.5
通讯作者：
Wang, Andrew
中科院分区：
文献类型：
--
作者：
Ren, Meiling;Xu, Hengyi;Huang, Xiaolin;Kuang, Min;Xiong, Yonghua;Xu, Hong;Xu, Yang;Chen, Hongyu;Wang, Andrew
采用CdSe/ZnS包封制备了高荧光量子点珠（QBs），并首次将其作为免疫层分析（ICA）信号放大探针用于玉米黄曲霉毒素B1 （AFB1）的超灵敏检测。使用高亮度量子点作为ICA探针的挑战是量子点的平滑流动和非特异性结合在硝化纤维素（NC）膜上，这是通过独特的聚合物封装量子点（QDs）和表面阻断方法来克服的。在最佳条件下，基于qb的ICA （QB-ICA）传感器对玉米提取物中AFB1的动态线性检测范围为5 ~ 60 pg mL-1，中位抑制浓度（IC50）为13.87±0.16 pg mL-1，显著低于QD作为信号探针的IC50(0.54±0.06 ng mL-1)，显著降低（39倍）。QB-ICA传感器对玉米提取物中AFB1的检出限（LOD）为0.42 pg mL-1，比先前报道的基于金纳米颗粒的免疫层析法（AuNP-ICA）提高了约2个数量级，甚至可以与传统的酶联免疫吸附法（ELISA）相媲美或更好。我们的QB-ICA传感器的性能和实用性通过商用ELISA试剂盒验证，并进一步通过液相色谱串联质谱（LC-MS /MS）验证。鉴于其有效的信号放大性能，所提出的QB-ICA为食品安全监测中分析物的快速、灵敏和经济的定量检测提供了巨大的潜力。
Highly luminescent quantum dot beads (QBs) were synthesized by encapsulating CdSe/ZnS and used for the first time as immunochromatographic assay (ICA) signal amplification probe for ultrasensitive detection of aflatoxin B1 (AFB1) in maize. The challenges to using high brightness QBs as probes for ICA are smooth flow of QBs and nonspecific binding on nitrocellulose (NC) membrane, which are overcome by unique polymer encapsulation of quantum dots (QDs) and surface blocking method. Under optimal conditions, the QB-based ICA (QB-ICA) sensor exhibited dynamic linear detection of AFB1 in maize extract from 5 to 60 pg mL–1, with a median inhibitory concentration (IC50) of 13.87 ± 0.16 pg mL–1, that is significantly (39-fold) lower than those of the QD as a signal probe (IC50 = 0.54 ± 0.06 ng mL–1). The limit of detection (LOD) for AFB1 using QB-ICA sensor was 0.42 pg mL–1 in maize extract, which is approximately 2 orders of magnitude better than those of previously reported gold nanoparticle based immunochromatographic assay (AuNP-ICA) and is even comparable with or better than the conventional enzyme-linked immunosorbent assay (ELISA) method. The performance and practicability of our QB-ICA sensor were validated with a commercial ELISA kit and further confirmed with liquid chromatography tandem mass spectrometry (LC–MS/MS). Given its efficient signal amplification performance, the proposed QB-ICA offers great potential for rapid, sensitive, and cost-effective quantitative detection of analytes in food safety monitoring.
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