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基于 SERS 的金纳米粒子靶向自组装用于苏云金芽孢杆菌转基因序列的放大检测

Target triggered self-assembly of Au nanoparticles for amplified detection of Bacillus thuringiensis transgenic sequence using SERS.

机构信息

College of Science, State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, PR China.

出版信息

Biosens Bioelectron. 2014 Dec 15;62:196-200. doi: 10.1016/j.bios.2014.06.046. Epub 2014 Jun 27.

Abstract

The research methods for DNA detection have been widely extended since the application of nanotechnology, but it remains a challenge to detect specific DNA sequences or low abundance genes in the biological samples with accuracy and sensitivity. Here we developed a SERS biosensing platform by target DNA (tDNA) triggered self-assembly of Au nanoparticles (Au NPs) probes on DNA nanowires for signal amplification in DNA analysis. Based on the hybridization chain reactions (HCR) and surface enhanced Raman scattering (SERS) technology, the SERS intensity reveals a good linearity with tDNA ranging from 50 pM to 500 pM under optimal conditions. The specific detection of tDNA sequence was realized with a detection limit of 50 pM (S/N=3). To demonstrate the specificity and universality of the strategy, the single-base mismatches in DNA and the Bacillus thuringiensis (Bt) transgenic sequence were successively applied in the SERS assay. The results showed that the sensitivity and accuracy of the SERS-based assay were comparable with real-time PCR. Besides, the method would provide precise and ultra-sensitive detection of tDNA but also informative supplement to the SERS biosensing platform.

摘要

自从纳米技术应用以来,DNA 检测的研究方法已经得到了广泛的扩展,但在生物样本中准确而灵敏地检测特定的 DNA 序列或低丰度基因仍然是一个挑战。在这里,我们通过目标 DNA(tDNA)触发 DNA 纳米线上的金纳米粒子(Au NPs)探针的自组装,开发了一种 SERS 生物传感平台,用于 DNA 分析中的信号放大。基于杂交链反应(HCR)和表面增强拉曼散射(SERS)技术,在最佳条件下,SERS 强度与 tDNA 呈良好的线性关系,范围从 50 pM 到 500 pM。通过单碱基错配在 DNA 中的特异性检测和苏云金芽孢杆菌(Bt)转基因序列的特异性检测,实现了 tDNA 序列的特异性检测,检测限为 50 pM(S/N=3)。为了证明该策略的特异性和通用性,我们将 DNA 中的单碱基错配和苏云金芽孢杆菌(Bt)转基因序列相继应用于 SERS 分析中。结果表明,基于 SERS 的测定法的灵敏度和准确性可与实时 PCR 相媲美。此外,该方法将为 tDNA 的精确和超灵敏检测提供信息补充,同时也为 SERS 生物传感平台提供了信息补充。

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