场效应晶体管(FET)生物传感器 2009

Poly-silicon nanowire field-effect transistor for ultrasensitive and label-free detection of pathogenic avian influenza DNA.

Biosensors & bioelectronics Lin CH, Hung CH, Hsiao CY, Lin HC, Ko FH, Yang YS
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组成图示

Poly-silicon nanowire field-effect tr... 传感器构成示意图

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传感器类型

场效应晶体管(FET)生物传感器

检测对象

高致病性禽流感病毒H5 DNA(H5 target DNA)、高致病性禽流感病毒H7 DNA(H7 target DNA);样品基质:PBS缓冲液(10 mM,pH 7.0)

检测原理

poly-SiNW FET表面先经APTES氨基化,再用戊二醛和NaBH3CN将互补捕获DNA探针偶联到纳米线表面。目标禽流感H5/H7 DNA与捕获探针特异性杂交后,DNA磷酸骨架将负电荷引入纳米线表面。对于n型poly-SiNW FET,表面负电荷改变沟道电场并降低载流子浓度,使漏极电流ID下降。杂交量随目标DNA浓度增加而增加,因此Δlog ID在1 fM–10 pM范围内随浓度升高,超过10 pM后趋于饱和。该检测为无标记检测,未使用酶或纳米酶放大,灵敏度主要来自纳米线高比表面积和FET场效应;栅压VG会影响响应幅度。

检测灵敏度

LOD: 1 fM;线性范围: 1 fM–10 pM

效应效果

未修饰poly-SiNW FET在PBS及H5/H7目标DNA中ID–VG曲线不变,仅APTES或APTES加戊二醛修饰也无响应,说明非特异结合低。cH7capture器件对H5target无响应,对10 pM H7target电流显著下降;cH5capture器件对H7target无响应,对10 pM H5target响应,选择性高。1 fM–10 pM呈浓度依赖,>10 pM饱和,100 pM仍稳定。与SPR(0.677 pM)和QCM(1 pM)无标记DNA检测相比,fM级灵敏度更优;与单晶SiNW FET性能相当但制备成本低、兼容商业半导体工艺、可大规模生产,作者认为适合便携式现场和即时诊断。

传感器的构成

  • 基底/换能器:N型多晶硅纳米线场效应晶体管(poly-SiNW FET),80 nm宽、2 μm长双纳米线通道,含源/漏电极与背栅,将表面电荷变化转换为漏极电流变化
  • 保护/绝缘层:100 nm湿氧化层(wet-oxide)和80 nm氮化层(nitride),防止液体侵入并降低栅漏电流
  • 表面化学修饰层:3-氨基丙基三乙氧基硅烷(APTES)引入氨基,戊二醛(glutaraldehyde)与氰基硼氢化钠(NaBH3CN)用于DNA探针偶联
  • 识别元件:5′-氨基修饰互补禽流感H5捕获DNA探针(cH5capture)或H7捕获DNA探针(cH7capture),特异性杂交目标DNA
  • 信号标记物:无标记(label-free);FITC仅用于功能化验证
  • 封闭剂:50 mM乙醇胺(ethanolamine),封闭未反应醛基,降低非特异结合
  • 样品/微流控层:PDMS/亚克力微流控通道与金属夹持器,维持PBS溶液包围纳米线
  • 信号读出:Keithley 2636芯片分析仪,测量ID–VG/ID–VD曲线,输出漏极电流变化

中文摘要

流感监测需要快速、稳健、低成本且能详细分析毒株的分析技术。本文报道了功能化多晶硅纳米线场效应晶体管(poly-SiNW FET)用于高致病性禽流感(AI)病毒H5和H7毒株DNA的特异性、超灵敏(飞摩尔级)检测。禽流感是重要传染病,亟需现场监测。该器件采用简单低成本方法制备,与现有商业半导体工艺兼容,无需昂贵电子束光刻工具,适合大规模生产。当poly-SiNW FET纳米线表面修饰互补捕获DNA探针后,加入禽流感病毒目标DNA可产生特异性电学响应;在fM至pM范围内可区分H5目标DNA。凭借优异电学性能和商业化量产潜力,poly-SiNW FET有望发展为用于现场检测和即时诊断的便携式生物传感器。

英文摘要

Enhanced surveillance of influenza requires rapid, robust, and inexpensive analytical techniques capable of providing a detailed analysis of influenza virus strains. Functionalized poly-crystalline silicon nanowire field-effect transistor (poly-SiNW FET) was demonstrated to achieve specific and ultrasensitive (at fM level) detection of high pathogenic strain virus (H5 and H7) DNA of avian influenza (AI) which is an important infectious disease and has an immediate need for surveillance. The poly-SiNW FET was prepared by a simple and low-cost method that is compatible with current commercial semiconductor process without expensive E-beam lithography tools for large-scale production. Specific electric changes were observed for AI virus DNA sensing when nanowire surface of poly-SiNW FET was modified with complementary captured DNA probe and target DNA (H5) at fM to pM range could be distinguished. With its excellent electric properties and potential for mass commercial production, poly-SiNW FET can be developed to become a portable biosensor for field use and point-of-care diagnoses.

关键词

多晶硅纳米线场效应晶体管禽流感DNA无标记检测生物传感器