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利用无标记金纳米粒子探针和动态光散射技术检测转基因基因片段的超灵敏方法。

An ultrasensitive method for the detection of gene fragment from transgenics using label-free gold nanoparticle probe and dynamic light scattering.

机构信息

College of Science, State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, PR China.

出版信息

Anal Chim Acta. 2011 Jun 24;696(1-2):1-5. doi: 10.1016/j.aca.2011.04.001. Epub 2011 Apr 13.

Abstract

The detection of transgenic products is of great significance for the development of transgenic technique. In this paper, we developed a simple, rapid and ultrasensitive method for the detection of sequence-specific Nopaline synthase (NOS) gene from the transgenic plants using label-free gold nanoparticle (NP) probe and dynamic light scattering (DLS) technology. Gold NPs were stable in NaCl solution with the presence of NOS gene probe. On the contrary, they were aggregated in NaCl solution when the probe sequence was hybridized with target sequence. The change in the size of gold NPs can be detected by DLS technology with high sensitivity. Under the optimal conditions, the average hydrodynamic diameter of gold NPs was linear with the concentration of the target sequence ranging from 1.0×10(-13)mol L(-1) to 5.0×10(-9)mol L(-1), with a detection limit of 3.0×10(-14)mol L(-1) (S/N=3). The relative standard deviation (at 1.0×10(-9)mol L(-1) of target sequence) was 4.8% (n=11). The result shows that gold NPs-based DLS method has great potential in the analysis of transgenic products.

摘要

转基因产品的检测对于转基因技术的发展具有重要意义。本文利用无标记金纳米粒子(NP)探针和动态光散射(DLS)技术,开发了一种简单、快速、超灵敏的检测转基因植物中序列特异性胭脂碱合成酶(NOS)基因的方法。在有NOS 基因探针存在的情况下,金 NPs 在 NaCl 溶液中稳定。相反,当探针序列与靶序列杂交时,它们在 NaCl 溶液中聚集。金 NPs 尺寸的变化可以通过 DLS 技术高灵敏度检测到。在最佳条件下,金 NPs 的平均水动力直径与靶序列的浓度呈线性关系,范围为 1.0×10(-13)mol L(-1)至 5.0×10(-9)mol L(-1),检测限为 3.0×10(-14)mol L(-1)(S/N=3)。在靶序列为 1.0×10(-9)mol L(-1)时的相对标准偏差(n=11)为 4.8%。结果表明,基于金纳米粒子的 DLS 方法在转基因产品分析中具有很大的潜力。

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