Ultrasensitive electrochemical biosensor for specific detection of DNA based on molecular beacon mediated circular strand displacement polymerization and hyperbranched rolling circle amplification

Using a cascade signal amplification strategy, an ultrasensitive electrochemical biosensor for specific detection of DNA based on molecular beacon (MB) mediated circular strand displacement polymerization (CSDP) and hyperbranched rolling circle amplification (HRCA) was proposed. The hybridization of MB probe to target DNA resulted in a conformational change of the MB and triggered the CSDP in the presence of bio-primer and Klenow fragment (KF exo⁻), leading to multiple biotin-tagged DNA duplex. Furthermore, the HRCA was implemented to product amounts of double-stranded DNA (ds-DNA) fragments using phi29 DNA polymerase via biotin-streptavidin interaction. After the product of HRCA binded numerous biotinylated detection probes, an ultrasensitive electrochemical readout by further employing the streptavidin-alkaline phosphatase. The proposed biosensor exhibited excellent detection sensitivity and specificity with a log-linear response to target DNA from 0.01 fM to 10 pM as low as 8.9 aM. The proposed method allowed DNA detection with simplicity, rapidness, low cost and high specificity, which might have the potential for application in clinical molecular diagnostics and environmental monitoring.     

核酸工具酶辅助的信号放大技术在生物分子检测中的应用

核酸工具酶具有催化能力高、序列识别能力强、反应条件温和以及生物相容性好等优势,因而被广泛地应用于核酸、蛋白质和小分子等生物活性分子的分析检测。本文主要对核酸工具酶进行介绍,并对应用于生物分子检测的一些核酸工具酶辅助的信号放大技术进行较为全面的综述,并展望了该技术的应用前景。     

Electrochemical Signal Tracing by Glucose Oxidase and Ferrocene Dually Functionalized Gold Nanoprobe for Ultrasensitive Immunoassay

A glucose oxidase (GOD) and ferrocene (Fc) dually functionalized gold nanoprobe was simply prepared for electrochemical immunoassay. By combination with sandwich immunoreaction at a carbon nanotube (CNT)‐based immunosensor and signal tracing of the nanoprobe through the Fc‐mediated GOD‐catalytic reaction, a new electrochemical immunoassay method was successfully developed. Both the multi‐enzyme signal amplification of the nanoprobe and the electron transfer promotion of the CNTs modified on the immunosensor greatly enhanced the signal response. Thus this method showed excellent analytical performance including ultrahigh sensitivity, wide linear range as well as good specificity, reproducibility, stability and reliability for human IgG measurement.     

基于无线通信的接触式带钢板形信号采集系统研制

针对滑环结构的接触式板形测试仪存在的不足,采用无线通信方式,研制了多单片机系统的嵌入式板形信号采集系统,通过光电接近开关的合理安装设计,巧妙解决了检测辊正反转时的信号采集和每组4个传感器信号区分问题,并给出了I2C总线系统中主、从单片机的流程。现场试验表明,在传感器为30组,检测辊10圈/秒转动时,系统的可靠性、数据传输速率和误码率均可满足板形闭环控制的要求。     

面向低强度无线网络信号接收谱参数估计研究

针对以北斗卫星导航信号为代表的亚纳秒级的低强度无线网络信号在定位中难以获取精确时间估计及角度估计,且易受环境噪声影响,使其定位精度不高等难题,本文提出了基于亚纳秒级的低强度无线网络信号接收谱参数估计方法。首先通过抽样方式,将发射信号抽样为多维独立子信号并独立建模,通过构造噪声空间与子信号空间在对应列向量正交化的基础上精确获取TOA估计;随后利用复数域映射,在获取TOA估计基础上采取比对方式精确地获取DOA估计。最后对本文参数估计方法进行了精度分析。测试数据显示：与 PM算法、ESPRIT算法相比,本文技术在TOA及DOA估计上更为精确;同时在信号强度低且背景噪声干扰严重的情况下,本文方法仍可有效的维持参数估计精度。     

一种多功能信号源系统设计

针对单一信号源无法满足测试需求的现状,提出了一种新型多功能信号源系统设计方案;详细阐述了信号源系统的硬件及软件设计,方案基于标准化、高速、可靠性三方面的考虑,硬件环境基于Compact PCI工控计算机,设计了3U CPCI结构的信号源板卡,配套软件采取模块化设计;该方案可实现多类型、多通道的高精度、高可靠性信号的独立输出和同时输出;实验结果表明,该多功能信号源系统稳定性好、可靠性高、使用便捷,满足设计需求。     

Investigation of several parameters influencing signal generation in flow-through membrane-based enzyme immunoassay

Rapid-response analytical tests that can be performed at the point of sampling are based on a visual detection system. The influence of different factors on the signal generation in a membrane-based enzyme immunoassay was investigated. The research was applied to a flow-through immunoassay for the detection of ochratoxin A (OTA). This assay format is a very convenient, simple and fast qualitative screening tool. Conjugates of OTA with horseradish peroxidase (HRP) and alkaline phosphatase (AP) were used as enzyme tracers. A new conjugate OTA-AP has been synthesized in our laboratory and its performance in the assay was compared with that of OTA-HRP. Different substrate systems for HRP and AP were compared. Several reagents, including polymers and surfactants, were tested for their possible effect on signal generation with the use of OTA-HRP conjugate. Polymers such as poly(vinyl alcohol) (PVA) and poly(ethylene glycol) (PEG) 6000 exerted a favourable effect on signal amplification, whereas surfactants negatively affected assay performance. The highest signal amplification (30–70% compared to the standard assay procedure) was achieved using 0.5% PVA in tetramethylbenzidine (TMB) Colorburst substrate solution and phosphate-buffered saline (PBS) for the washing step. It allowed more reliable visual estimation of the results from OTA-HRP assay. Exclusion of the detergent (Tween 20) from the washing solution exerted a favourable effect on assay performance using both enzyme tracers. The assay using OTA-HRP was more susceptible to matrix interferences than the assay with OTA-AP. Signal development in the matrix was better for the OTA-AP assay and visual estimation of the results was easier to perform in this case. For the analysis of spiked wheat samples, OTA-AP conjugate gave a more sensitive, stable and reproducible assay with a cut-off level of 4 μg kg⁻¹ for OTA. The application of the new OTA-AP conjugate resulted in improved assay performance for the food samples.     

MicroRNAs电化学生物传感器在乳腺癌检测中的研究进展

MicroRNAs是一类内源性非编码小RNA分子,可调控靶基因的表达。特异性microRNAs的失调在诸如癌症、心血管疾病、免疫疾病、神经退行性疾病和皮肤疾病等的发展过程中起着关键作用,常作为疾病早期诊断和预后的生物标志物。电化学生物传感器由于其灵敏、快速、成本低等优势,已经成为传统microRNAs检测方法的一种很有前途的替代方法。本文综述了基于microRNA电化学生物传感器检测乳腺癌的研究进展,并对其所构建的电化学生物传感器的检测方法进行了分析探讨。     

Natural Product Synthesis on Polymeric Supports—Synthesis and Biological Evaluation of an Indolactam Library

Potent activators of protein kinase C in fibroblasts: This property was determined for several indolactam V analogues ( 1 ) with a new cell-based assay system. This tumor-promoting indole alkaloid and analogues thereof can be synthesized efficiently on the solid phase. The key steps of the combinatorial approach are a regioselective amination of the indole ring and an enantioselective enzymatic reaction.     

基于Hilbert变换的水下多源声信号频率的相干探测

为实现水下中低频声信号的探测识别,通过研究水下多声源相干探测信号的特征,理论上给出了相干探测信号频谱混叠情况下的特征表达式,并提出了一种基于Hilbert变换的信号解调处理方法,实现了水下多声源相干探测信号频谱混叠情况下各声源发声频率的解调.该方法将探测信号经过滤波平滑处理之后进行Hilbert变换,得到信号的解析形式,然后对解析信号模值的平方进行二次滤波平滑等处理,分离混叠在一起的频带,将得到的信号进行频谱分析,根据频移值计算得到水下各个声源的发声频率.在光学暗室下搭建激光相干探测系统,对2~6kHz的水下声信号进行实验,实验结果表明,该方法可以有效分离探测信号中混叠在一起的信号频带,并准确提取各水下声信号的发声频率,频率提取重复性不大于2.5Hz.