基于DNA三链体结构和核酸外切酶III辅助循环扩增的三聚氰胺均相电化学生物传感器,Analytical Chemistry

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基于DNA三链体结构和核酸外切酶III辅助循环扩增的三聚氰胺均相电化学生物传感器
Analytical Chemistry ( IF 6.7 ) Pub Date : 2016-10-03 00:00:00 , DOI: 10.1021/acs.analchem.6b02753
Caili Fu , Chang Liu , Ying Li , Yajing Guo , Fang Luo , Peilong Wang ₁ , Longhua Guo , Bin Qiu , Zhenyu Lin

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三元结构中的无碱基位点（AP位点）可以高选择性地识别特定的靶标。在这项研究中，此特征首先用于开发简单，灵敏和选择性的均相电化学生物传感器，用于三聚氰胺的测定。该测定法具有以下优势：DNA三链体结构对AP具有三聚氰胺选择性，对三聚氰胺具有高选择性，并且核酸外切酶（Exo）III辅助的循环扩增效率高。精心设计了DNA-1（T1），DNA-2（T2），含有AP位点的poly [dA]探针（8A）和亚甲基蓝标记的DNA探针（dMB探针）。三聚氰胺可以通过胸腺嘧啶和三聚氰胺之间的氢键相互作用特异性地定位在AP位点，从而使T1，T2和8A彼此靠近，因此形成稳定的T-三聚氰胺-T DNA三链体结构。在最佳条件下微分脉冲伏安（DPV）响应与三聚氰胺浓度的对数在1 nM〜0.5μM范围内呈线性关系。研发的生物传感器已成功应用于从三聚氰胺碗中检测三聚氰胺的迁移。结果表明，在4％乙酸溶剂中的迁移最大，这与高效液相色谱检测的结果相似。这种均质的电化学传感器在检测乳制品中的三聚氰胺和多类食品包装或应用材料中三聚氰胺的迁移方面可能具有潜在的前景。研发的生物传感器已成功应用于从三聚氰胺碗中检测三聚氰胺的迁移。结果表明，在4％乙酸溶剂中的迁移最大，这与高效液相色谱检测的结果相似。这种均质的电化学传感器在检测乳制品中的三聚氰胺和多类食品包装或应用材料中三聚氰胺的迁移方面可能具有潜在的前景。研发的生物传感器已成功应用于从三聚氰胺碗中检测三聚氰胺的迁移。结果表明，在4％乙酸溶剂中的迁移最大，这与高效液相色谱检测的结果相似。这种均质的电化学传感器在检测乳制品中的三聚氰胺和多类食品包装或应用材料中三聚氰胺的迁移方面可能具有潜在的前景。

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Homogeneous Electrochemical Biosensor for Melamine Based on DNA Triplex Structure and Exonuclease III-Assisted Recycling Amplification

Abasic site (AP site) in the triplex structure can recognize specific target with high selectivity. In this study, this character was first applied to develop a simple, sensitive, and selective homogeneous electrochemical biosensor for melamine determination. The assay combines the advantage of the high selectivity of the DNA triplex structure containing an AP site to melamine and high efficiency of exonuclease (Exo) III-assisted recycling amplification. DNA-1 (T1), DNA-2 (T2), poly[dA] probe containing an AP site (8A) and methylene blue-labeled DNA probe (dMB probe) were carefully designed. Melamine can specifically locate in the AP site through hydrogen bonding interaction between thymine and melamine to make T1, T2, and 8A close to each other, therefore, forming a stable T-melamine-T DNA triplex structure. Under the optimal conditions, the differential pulse voltammetric (DPV) response had a linear relationship with the logarithm of melamine concentration in the range of 1 nM∼0.5 μM. The developed biosensor has been successfully applied to detect the migration of melamine from melamine bowl. Result showed that the migration in 4% acetic acid solvent was the largest, which is similar to that detected by high performance liquid chromatography. This homogeneous electrochemical sensor may have a potential prospect in detecting melamine in dairy products and migration of melamine from multicategory food packaging or application materials.

更新日期：2016-10-03

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