传感器类型
表面等离子共振(SPR)生物传感器
检测对象
鱼精DNA(fish sperm DNA, fsDNA);样品基质为H3PO4–KH2PO4缓冲液及含金属离子、葡萄糖、BSA的合成样品
检测原理
金纳米棒表面吸附CTAB形成正电荷界面,在pH 2.2酸性缓冲液中,带负电的fsDNA通过静电吸附结合到AuNRs表面。DNA结合改变纳米棒周围局部折射率,并诱导AuNRs发生端-端和侧-侧聚集,使纳米棒间距离缩短,产生局域表面等离子体耦合。聚集导致纵向LSPR带红移、吸收峰变宽,同时散射体尺寸增大和电子耦合增强共振光散射(RLS)强度。在581 nm处监测增强RLS强度ΔIRLS,其随fsDNA浓度增加而增大,实现无标记定量检测。
检测灵敏度
线性范围: 0.05–0.5 μg/mL;灵敏度斜率: 7.204;回归方程: ΔI = 9.946 + 7.204C;相关系数: 0.998
效应效果
该方法对多种金属离子、糖和蛋白质具有较好抗干扰性:Na+(500 μM)、K+(200 μM)、Ca2+(100 μM)、Mg2+(100 μM)等对RLS信号影响较小,变化在-8.4%至+11.7%范围内;葡萄糖(1 μM)和BSA(0.1 μg/mL)影响分别为+12.5%和+7.5%。合成样品分析中,含金属离子样品回收率为103.8%,含葡萄糖和BSA样品回收率为94.2%,RSD分别为2.4%和2.8%(均小于2.8%)。方法无需有机荧光染料或放射性标记,具有简单、快速、灵敏、稳定等优点,适用于实际样品中DNA的测定。
传感器的构成
- 光学换能器:金纳米棒(AuNRs),提供LSPR与RLS信号,平均长度约58±5 nm、直径约26±3 nm
- 表面修饰层:十六烷基三甲基溴化铵(CTAB)双分子层,吸附于AuNRs表面使其带正电并稳定纳米棒
- 识别/结合界面:CTAB正电荷表面,与带负电的fsDNA发生静电吸附,无独立特异性识别元件
- 缓冲介质:H3PO4–KH2PO4缓冲液(pH 2.2,5.0 mmol/L),维持酸性环境并控制离子强度
- 信号读出:共振光散射(RLS)光谱仪,在581 nm监测增强RLS强度ΔIRLS
中文摘要
本文报道了一种基于金纳米棒局域表面等离子共振(LSPR)的共振光散射(RLS)光谱法用于DNA定量检测。金纳米棒作为光学换能器,其表面吸附阳离子表面活性剂CTAB而带正电,可与带负电的DNA通过静电作用结合。DNA结合引起金纳米棒周围界面折射率变化及纳米棒聚集,导致RLS强度增强,增强峰位于311 nm和581 nm,其中581 nm处信号随DNA浓度增加而增强。在优化条件下,增强RLS强度与DNA浓度在0.05–0.5 μg/mL范围内呈线性关系。该方法无需有机荧光染料或放射性标记,具有简单、快速、灵敏和稳定等优点,并成功应用于合成样品中DNA的测定,结果令人满意。
英文摘要
A resonance light scattering (RLS) spectrometry for determining DNA based on localized surface plasmon resonance (LSPR) of gold nanorods was developed. The fabrication and characterization of the biosensor utilized gold nanorods as the optical transducer was described. The DNA was determined based on the binding of DNA to gold nanorods by electrostatic adsorption. The RLS intensity of the gold nanorods was enhanced in the presence of DNA. The peaks of the enhanced RLS spectrum were at the wavelength 311 and 581 nm. The binding of DNA to the gold nanorods causes an increase of the RLS intensity that is responsive to the concentration of DNA. Under the optimal conditions, the enhanced RLS intensities are directly proportional to the DNA concentrations in the range of 0.05-0.5 microg/mL. The proposed method was successfully applied to the determination of DNA in the samples and the results were satisfactory.