A Novel Fluorescent FRET Hairpin Probe Switch for aflD Gene Detection in Real Fermented Soybean Paste

文献类型： 外文期刊

第一作者： Li, Yaqi

作者： Li, Yaqi;Li, Jiyang;Kong, Dezhao;Shi, Qiaoqiao;Liu, Chang;Yu, Tao;Dong, Chunwang

作者机构：

关键词： FRET switch; Hairpin probe; Quantum dots; AuNPs; aflD genes

期刊名称：FOOD ANALYTICAL METHODS （ 影响因子：3.366； 五年影响因子：3.07 ）

ISSN： 1936-9751

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收录情况： SCI

摘要： The development of detection method for aflatoxin gene cluster in Aspergillus flavus and Aspergillus parasiticusis is an effective approach to prevent the widespread of aflatoxins, which are considered as extremely, toxic substances. Herein, a hairpin DNA-assisted fluorescence probe switch based on Forster resonant energy transfer (FRET) was designed. In this work, the two terminals of hairpin DNA probe were combined with CdTe/CdS quantum dots as donor and gold nanoparticles (AuNPs) as acceptor respectively to fabricate the FRET hairpin probe switch, which were utilized to detect the aflatoxigenic fragment of aflD genes for the first time. In the absence of target DNA, the hairpin probe formed the double stem loops state to trigger the FRET to induce the fluorescence-quenching (turning off). With the addition of target DNA, the specific interaction between target and hairpin probe switch brought about the separation of fluorescent energy donor-acceptor pairs to create an increasing fluorescence response (turning on). Consequently, the fluorescence intensity recovers can be employed to detect aflD genes in homogenous phase through optical responses. This proposed FRET hairpin probe switch performed a great limit of detection of 0.02 nM with a linear range of 0.05-200 nM under optimal conditions. Furthermore, the feasibility of the fabricated strategy was demonstrated in fermented soybean paste with the advantages of simplicity, rapidity, and sensitivity, which indicated that this method exhibited the potential and practicability in foodborne biological samples.

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