综述或非传感器论文 2009 非传感器论文

Recent development of nano-materials used in DNA biosensors.

Sensors (Basel, Switzerland) Xu K, Huang J, Ye Z, Ying Y, Li Y
阅读原文 PDF DOI PubMed

组成图示

Recent development of nano-materials ... 传感器构成示意图

点击图片查看大图 · 依据论文自动绘制

传感器类型

综述或非传感器论文

检测对象

特定DNA序列(target DNA/oligonucleotide);样品基质包括生物样品、食品、环境水样、细胞/染色体DNA等

检测原理

DNA生物传感器通常将序列特异性ssDNA探针或PNA探针固定在纳米材料修饰的电极、SPR芯片或石英晶体表面。目标DNA与探针杂交后,双链形成引起界面电荷、质量、电子转移阻抗或光学性质变化。纳米材料一方面作为高比表面积基底提高探针负载量,另一方面作为Au/Ag/CdS纳米颗粒或CNT载体标记物,通过质量增加、电化学活性增强、SPR场增强或酶催化产物积累实现信号放大。信号随目标DNA浓度升高而增强或发生可测偏移,经伏安、EIS、SPR、QCM、比色或ECL读出。

检测灵敏度

综述报道多个代表性数值:LOD: 10 fM;LOD: 1.38 fM;LOD: 0.2 pmol/L;LOD: 40 pg/mL;LOD: 2.3 × 10−13 mol/L;线性范围: 1.0 × 10−12 mol/L 至 1.0 × 10−7 mol/L;检测范围: 1 × 10−1 至 1 × 10−5 μM

效应效果

综述显示纳米材料可显著提升DNA传感器性能。CNT修饰玻碳电极使双链DNA鸟嘌呤氧化信号比裸电极高20倍,五个电极RSD为3.4%;CNT负载CdS标记使检测限较单颗粒剥离杂交降低约500倍,250倍非互补寡核苷酸干扰很小;CNT-HRP标签比传统ELOSA标签灵敏度至少提高1000倍;Au纳米簇嵌入SiO2的SPR分辨率较常规SPR提高10倍;金刚石纳米线传感器灵敏度达2 pM,并可稳定经历30次杂交/变性循环。作者认为小尺寸纳米材料及多种纳米材料组合可改善灵敏度、选择性和稳定性,未来有望实现无PCR的DNA检测。

传感器的构成

  • 基底/换能器:金电极、玻碳电极(GCE)、ITO、硅/蓝宝石、石英晶体、SPR芯片,提供电学/光学/压电换能基础
  • 纳米修饰层:Au NPs、Pt NPs、ZrO2、Pr6O11、SiO2、CNTs、聚吡咯(PPy)、聚苯胺、壳聚糖,扩大表面积并固定探针
  • 识别元件:单链DNA探针(ssDNA)、寡核苷酸(ODN)、肽核酸(PNA),与目标DNA序列特异性杂交
  • 信号标记物:Au NPs、Ag NPs、CdS NPs、CNT负载CdS或Ru复合物、HRP、链霉亲和素-生物素,放大杂交信号
  • 抗非特异吸附层:羧基化葡聚糖膜(SPR体系),减少ODN/Au-NP非特异吸附
  • 换能读出:伏安法、EIS、SPR、QCM、比色、ECL、电阻测量,将界面变化转为可测信号

中文摘要

随着对核酸分子结构与功能认识的深入,序列特异性DNA检测日益重要。基于核酸杂交的DNA生物传感器因特异性高、速度快、便携且成本低而受到积极开发。近年来,纳米材料在DNA生物传感器中的应用备受关注。由于具有高比表面积和良好生物相容性,纳米材料可增加DNA探针固定量,且结合在纳米材料上的DNA能保持生物活性;另一方面,用纳米材料标记目标分析物也可实现信号放大。本综述总结了近五年各类纳米材料在DNA生物传感器中的应用。作者发现,小尺寸纳米材料更适合作为DNA固定基底或信号放大标记物;在传感器中使用两种或多种纳米材料可提升整体性能并弥补单一纳米组分的不足。目前多数DNA生物传感器仍需PCR扩增,但开发更小尺寸和/或生物化学性能更优的纳米材料将显著提高其准确性、选择性和灵敏度,无PCR的DNA生物传感器有望在可预见的未来实现。

英文摘要

As knowledge of the structure and function of nucleic acid molecules has increased, sequence-specific DNA detection has gained increased importance. DNA biosensors based on nucleic acid hybridization have been actively developed because of their specificity, speed, portability, and low cost. Recently, there has been considerable interest in using nano-materials for DNA biosensors. Because of their high surface-to-volume ratios and excellent biological compatibilities, nano-materials could be used to increase the amount of DNA immobilization; moreover, DNA bound to nano-materials can maintain its biological activity. Alternatively, signal amplification by labeling a targeted analyte with nano-materials has also been reported for DNA biosensors in many papers. This review summarizes the applications of various nano-materials for DNA biosensors during past five years. We found that nano-materials of small sizes were advantageous as substrates for DNA attachment or as labels for signal amplification; and use of two or more types of nano-materials in the biosensors could improve their overall quality and to overcome the deficiencies of the individual nano-components. Most current DNA biosensors require the use of polymerase chain reaction (PCR) in their protocols. However, further development of nano-materials with smaller size and/or with improved biological and chemical properties would substantially enhance the accuracy, selectivity and sensitivity of DNA biosensors. Thus, DNA biosensors without PCR amplification may become a reality in the foreseeable future.

关键词

DNA生物传感器纳米材料信号放大电化学检测表面等离子共振碳纳米管