DNA电化学传感器的研制

利用自组装单分子膜技术将巯己基修饰的单链DNA固定在金电极表面，以电活性的Hoechst 33258为指示剂，考察了单链DNA修饰电极（ssDNA/Au电极）双连DNA修饰电极（dsDNA/Au电极）的指示剂氧化峰电流，并利用其差值（Δip)与互补DNA浓度成线性关系对待测DNA浓度进行定量，因此，DNA电化学传感器可用于特定DNA序列的识别和测定。     

基于特殊形貌CdS纳米颗粒修饰的DNA传感器在DNA杂交信号检测中的应用

用水热法制备出具有特殊核桃状外表的纳米小球修饰在玻碳电极的表面,通过5′端巯基修饰的探针DNA共价结合在CdS层敏感层上形成共聚物,再与靶DNA杂交,利用循环伏安法(CV)和差分脉冲伏安法(DPV)研究修饰电极的电化学行为。修饰CdS纳米颗粒的电极检测得到的DNA杂交信号有明显的增强,峰电流强度值与靶DNA浓度值的负对数具有较好的线性关系,信号增强的最大值在靶DNA浓度为101μmol/L时得到。传感器灵敏度提高,检测下限可达1pmol/L以下。     

基于二茂铁标记的电化学DNA生物传感器研究

将二茂铁和巯基修饰的脱氧核糖核酸(DNA)探针通过自组装的方式固定到金电极表面,以二茂铁作为电子介体构建电化学DNA传感器,利用杂化前后DNA传感器所展现出峰电流的差异,实现对目标DNA的定量检测。通过研究DNA杂化前后方波伏安法的扫描频率与二茂铁电子介体传输速率关系,优化扫描频率。结果表明:当扫描频率200 Hz时,目标DNA浓度在5. 0×10~(-9)～1. 0×10~(-7)mol/L范围内,峰电流的变化与目标DNA浓度呈线性关系,线性拟合方程式为ΔI_P/I_0(%)=0. 760 95+0. 610 65 c,检测限为1. 7×10~(-9)mol/L(S/N=3)。     

基于树状高分子的DNA电化学传感器对禽流感病毒的检测

将G4 PAMAM固定在玻碳电极表面,然后通过共价作用固定禽流感病毒探针ssDNA-1,以[Co(phen)3]3+为指示剂,采用示差脉冲伏安法和交流阻抗法对DNA电化学生物传感器进行了表征.结果发现,通过与双链dsDNA作用的[Co(phen)3]3+的峰电流信号的变化,可以识别和定量检测溶液中互补的禽流感病毒DNA片段.经过条件优化,该法测定DNA的浓度线性范围为1.3×10-9～6.5×10-8 mol/L,最低检测限为9.2×10-10 mol/L.     

基于NaOH刻蚀玻碳电极的电化学传感器在检测膀胱癌DNA中的应用

制备了一种基于活化的玻碳电极的新型电化学DNA生物传感器,可用于膀胱癌DNA的检测.通过循环伏安法(CV)实现玻碳电极在NaOH溶液中的刻蚀,使电极表面负载大量官能团,为DNA提供连接位点,由Laviron方程计算得到玻碳电极表面的羧基浓度为 1.022×10-6 mol/cm2.亚甲基蓝(MB)作为电化学检测的杂交指示剂.采用原子力显微镜(AFM)对刻蚀后的电极进行了形貌表征.在最优杂交条件下,通过差分脉冲法(DPV)计算出最佳检测限为5.677×10-13 mol/L(n=5),适用目标 DNA浓度范围1×10-8 mol/L~1×10-12 mol/L.该传感器有望用于实际样品中膀胱癌DNA的快速检测.     

碲化钴纳米管/纳米金/壳聚糖复合膜DNA电化学传感器的构建

通过水热法制备了碲化钴纳米管,以制备的碲化钴纳米管、纳米金和壳聚糖组成的复合膜修饰裸铂电极,构建了一新型DNA电化学生物传感器.利用该生物传感器及邻菲罗啉钴电化学杂交指示剂对禽病毒基因进行了检测研究.实验表明,该法测定DNA的浓度线性范围为2.0×10-10～2.0×10-6 mol/L,最低检测限为7.94× 10-11 mol/L.     

种属DNA伏安传感器的制备及其应用研究

在羧酸修饰的碳糊电极表面共价键合人ssDNA,制备了人DNA伏安传感器.在杂交液中,传感器表面上的人ssDNA与杂交液中的人或动物ssDNA进行杂交反应时,电活性物质Co(bpy)3(ClO4)3配合物嵌入DNA双链中,使峰电流增加(△ip).△ip与试液中DNA浓度成正比,可用于检测生化样品中DNA含量,线性范围为0.010～0.20 mg/L,检测下限为2.29цg/L.     

基于氧化石墨烯/金纳米粒子放大电信号的二茂铁标记型溶菌酶适体传感器

设计了一种简单的信号衰减型电化学溶菌酶适体传感器。以氧化石墨烯修饰的玻碳电极为基底（GO/GCE）,通过循环伏安法电沉积金纳米粒子在电极表面,得到AuNPs/GO/GCE。将3’端修饰巯基的单链DNA通过金硫键共价连接于电极上,而在与前者可部分杂交的适体探针链5’端标记上二茂铁甲酸。当目标物溶菌酶与适体探针链发生特异性结合,通过示差脉冲伏安法检测到二茂铁甲酸还原峰电流相应的改变,从而实现对溶菌酶的识别及定量检测。结果表明,该传感器对溶菌酶具有较好的选择性和灵敏度,对于溶菌酶的检测范围为1.08×10-11～1.08×10-8mol/L,检出限达到6.02×10-12mol/L。     

基于“树枝状”信号放大的电化学DNA生物传感器研究

发展了一种基于"树枝状"信号放大的电化学生物传感器用于DNA的检测。该传感器利用两种DNA功能化的纳米金颗粒,通过两次"三明治"杂交,在电极表面形成"树枝"状结构,从而实现DNA的定量检测。首先通过共价交联方法获得巯基DNA1和DNA2修饰的两种纳米金颗粒,其中DNA1和DNA2与目标cDNA部分互补。然后,修饰在金电极上的捕获探针DNA1与目标cDNA分子及巯基DNA2修饰的纳米金颗粒(DNA2-AuNPs)形成第一个"三明治"杂交结构,实现一次放大检测。接着,DNA2-AuNPs又可与cDNA、巯基DNA1修饰的纳米金颗粒(DNA1-AuNPs)形成第二个"三明治"杂交结构,实现二次放大检测。这种"树枝状"放大信号的方法的检测限是0.13pmol/L,相对仅利用纳米金颗粒放大的方法而言,其检测限降低了4倍。并且,该传感器具有较好的识别碱基错配的能力。     

用于液相检测的双参数压电DNA传感器研究

采用自组装技术,利用亲和素-生物素系统将25-mer的生物素标记的DNA探针固定于石英谐振器金电极上,与双参数压电传感器相结合,研制成了用于液相检测的压电DNA传感器。该传感器稳定性较好,3h频移在5Hz以内;液体的体积在一定范围内对双参数传感器未见明显影响;用双参数方法制作的传感器特异性等性能较好,为实现检测的自动化和生物的动力学测定打下了基础。     

电化学沉积二氧化锆制备DNA电化学传感器

设计了一种简单、方便的电化学DNA生物传感器的制作方法,首先在硼掺杂的金刚石电极(BDD)表面电化学沉积一层二氧化锆薄膜,利用二氧化锆的分子结构特点,将探针DNA即5’末端修饰磷酸基团的寡核苷酸短链(ssDNA)连接到二氧化锆薄膜上.以亚甲基蓝(MB)为氧化还原的电子媒介体,应用循环伏安法和差分脉冲法测试了该电极的性能...     

硅烷法固定ssDNA制作的压电式DNA传感器

利用硅烷法将ssDNA固化T方向切割、3.58MHz和10MHz石英晶体金电极上,构建了压电DNA传感器,建立了快速检测葡萄球菌肠毒素B(SEB)基因的方法。优化了固定SEB基因的条件,并用动态法、静态法和斑点杂交法进行了固化效果观察。结果表明:该传感器2h后,有较好响应;对不同长度DNA探针固化效果进行比较,169bp(即169个碱基)SEB探针固定和杂交效果较好;利用0.5mol/LNaOH进行电极洗脱,时间为45min,电极可以再生,且传感器可以重复使用3次;利用1.2mol/LNaOH和1.2mol/LHCl洗涤电极,该电极可多次再利用;固化好的电极于4℃条件下可以保存三周之久。     

Development of an SH-SAW sensor for the detection of DNA hybridization

We have developed SH (shear horizontal) surface acoustic wave (SAW) sensors for the detection of DNA (deoxyribonucleic acid) hybridization on the gold-coated delay line of transverse SAW devices. DNA hybridization experiments were performed with 15-mer oligonucleotides (probe and complementary target DNA). The sensor consists of twin SAW delay line oscillators (sensing channel and reference channel) operating at 100 MHz fabricated on 36° rotated Y-cut X-propagation LiTaO₃ piezoelectric single crystals. The relative change in the frequency of the two oscillators was monitored to detect hybridization between the target DNA and probe DNA immobilized on the delay line of the SAW sensor. The measurement results showed a good response of the sensor to the mass loading effects of the DNA hybridization with a sensitivity level up to 1.55 ng/ml/Hz.     

毛细管DNA传感器

研究一种新型的利用毛细管作为载体检测靶DNA的生物传感器的可行性。首先应用0.2%多聚赖氨酸在毛细管内壁固定1×10-4mol/L ssDNA,与3.33×10-5mol/L靶DNA杂交后,应用1 g/L EB染色,在RF5000荧光仪上进行检测,t=5.5629,p<0.05,测定组与试剂空白组的荧光强度有明显差别;通过对12 h和14 h的杂交时间的比较,14 h的荧光强度明显高于12 h;对0.01,0.1,1,100,1 000 nmol/L的靶DNA进行测试,随靶DNA浓度的增高,荧光强度增强。实验结果表明:在光学DNA传感器的构建中,毛细管作为探针固定和荧光检测的载体是完全可行的。     

压电基因传感器检测芽孢杆菌靶序列研究

在石英晶体微天平(QCM)上用生物素亲和素法和自组装法2种不同的方法固定寡核苷酸探针,构建压电基因传感器,对芽孢杆菌靶序列进行实时检测。结果表明:生物素亲和素法固定探针效果更好,靶序列浓度为0.05~0.5μmol/L时,有很好的线性关系,线性回归方程为Y=93.88X+10.88(R=0.989 0,N=6,P<0.001),非线性误差为±4.7%。该传感器特异性较好,能够识别错配3个碱基的序列。     

一种工业用微型溶氧传感器的研制

以聚四氟乙烯和纯金电极为主要材料,制成了一种成本低、操作简便的Clark型溶氧传感器,并将该传感器在不同温度下的淡水和盐水中分别进行了测试。传感器的响应电流和溶解氧浓度之间呈良好的线性关系,在溶解氧浓度0～8mg/L范围内,误差小于±0.4mg/L。     

Development of an integrated CMOS DNA detection biochip

This paper presents a CMOS DNA detection biochip using an electrical detection method with self-assembly multilayer gold nanoparticles (AuNPs). Each measuring spot of this biochip consists of three major parts; a pair of electrodes with a nanogap, a current amplifier circuit, and a heater with an embedded temperature sensor. The biochip is first fabricated by a TSMC (Taiwan Semiconductor Manufacturing Company Ltd.) 0.35 μm 2P4M standard CMOS process. Then, post-CMOS micromachining etch processes are used to expose the surface of the nanogap to test samples for the establishment of multilayer AuNPs through hybridization between single strand DNAs in the samples. The gap distance between a pair of electrodes is 350 nm. Before taking DNA detection measurements, self-assembly monolayer AuNPs is established on the nanogap surface between two microelectrodes. Multilayer AuNPs can be observed if hybridization between single strand DNAs occurs. An approximately 1000-fold increase in electric current between the multilayer AuNPs over the monolayer AuNPs serves an indication of the presence of target DNA in test samples. After integrating the electrodes with an embedded current amplifier, the electric current of multilayer AuNPs is amplified to the order of mA that can be easily measured by a commercial Volt-Ohm-Milliammeter. The heating system with a heating element and a temperature sensor can be used to distinguish single base-pair mismatch hybridization from complementary hybridization for the establishment of multilayer AuNPs. The lowest detectable concentration of target DNA on this biochip is 0.1 nM.     

Preparation of an electrochemical PNA biosensor for detection of target DNA sequence and single nucleotide mutation on p53 tumor suppressor gene corresponding oligonucleotide

Development of an electrochemical biosensor based on peptide nucleic acid (PNA) probe for detection of target DNA sequence and single nucleotide mutation in p53 tumor suppressor gene corresponding oligonucleotide using methylene blue (MB) as an electrochemical indicator is described. The interaction between MB and short sequence of p53, one of the most important tumor suppressor genes due to its dysfunction in the majority of human cancers, was studied by differential pulse voltammety (DPV). Probe modified electrode was prepared by self-assembled monolayer (SAM) formation of thiolated PNA molecules on the surface of gold electrode (GE). The hybridization of PNA probe with target DNA was performed in solution to form PNA-DNA hybrid on the surface of the GE. A significant increase in the reduction signal of MB was observed upon hybridization of the probe with the complementary DNA. The selectivity of the biosensor was studied using noncomplementary oligonucleotides. Furthermore, our results confirmed the ability of the sensor to detect single base mismatch in the sample oligonucleotide. The influence of probe concentration on the effective discrimination against noncomplementary sequence and point mutation was also investigated. Diagnostic performance of the biosensor is described and the detection limit is found 6.82 × 10⁻¹⁰ M. The electrochemical impedance spectroscopy was also employed to further investigate the sensor function.