基于内含肽技术的生物传感器

开发用于分析复杂生物学问题的生物传感器，对于研究人类疾病和发现潜在药物分子十分重要。内含肽介导的蛋白质剪接技术的应用推动了新型生物传感器发展，基于内含肽技术的生物传感器可以对酶活性、激素受体、维生素和蛋白质与蛋白质间的相互作用等一些重要的生物学过程进行高效精准的分析和研究。重点阐述基于内含肽技术的生物传感器相关研究进展，并对其进行了展望。     

用于农药残留快速检测的生物传感器灵敏度筛选试验(简报)

为了提高乙酰胆碱酯酶生物传感器的灵敏度,使其更有效地应用于有机磷农药的快速检测。以商品乙酰胆碱酯酶（家蝇）和自制的鸡肝酶、鸡脑酶、鲤鱼脑酶为研究对象,比较了这4种乙酰胆碱酯酶的活性;检测了不同浓度的氧乐果、敌百虫和敌敌畏对4种酶的酶活抑制率。结果表明,鸡脑乙酰胆碱酯酶活性抑制率最高,3种农药对其的抑制率分别为：敌敌畏100 μg/L的抑制率为54.1%,敌百虫100 μg/L的抑制率为50.56%,氧化乐果100 μg/L的抑制率为24.16%。鸡脑酶廉价易得,且易于纯化,与自制的鸡肝酶和鲤鱼脑酶相比,乙酰胆碱酯酶的活性抑制率显著较高。因此鸡脑酶可以作为农产品农药残留快速检测用酶。     

应用SPR生物传感器检测酸菜发酵液中大肠杆菌

应用SPR传感器可以快速检测酸菜发酵液中的大肠杆菌,以便检测和控制酸菜发酵液中的亚硝酸盐含量,保证发酵食品安全。采用生物素-亲和素系统增大SPR传感器折射信号,获得了传感器的折射率与大肠杆菌浓度的回归方程,实现了对酸菜发酵液中大肠杆菌浓度快速,精确检测,提高大肠杆菌检测限到104cfu·mL-1。     

布拉酵母菌海藻糖提取方法的比较研究

本试验对布拉酵母菌海藻糖提取方法做了初步研究,通过3种提取剂,采用单一及复合处理方法,摸索出布拉酵母菌海藻糖提取的最佳方法是以5%的三氯乙酸做提取剂,超声破碎10 min后100℃水浴30 min。提取后用5%盐酸水解提取液,然后用生物传感器测定葡萄糖含量,计算海藻糖含量。     

Silencing of Erwinia amylovora sy69 AHL‐quorum sensing by a Bacillus simplex AHL‐inducible aiiA gene encoding a zinc‐dependent N‐acyl‐homoserine lactonase

Quorum sensing in Gram‐negative bacteria is regulated by diffusible signal molecules called N‐acyl‐l ‐homoserine lactones (AHLs). These molecules are degraded by lactonases. In this study, six Bacillus simplex isolates were characterized and identified as a new quorum‐quenching species of Bacillus. An aiiA gene encoding an AHL‐lactonase was identified based on evidence that: (i) it showed high homology with other aiiA genes of Bacillus sp.; (ii) the deduced amino acid sequence contained two conserved regions, ¹⁰⁴SHLHFDH¹¹¹ and ¹⁶⁵TPGHTPGH¹⁷³, characteristic of the metallo‐β‐lactamase superfamily; and (iii) the protein had zinc‐dependent AHL‐degrading activity. Additionally, the expression of the aiiA gene was significantly up‐regulated by 3‐oxo‐AHL. The AHL‐lactonase inhibited multiplication of the 3‐oxo‐C6‐AHL‐producing plant pathogen Erwinia amylovora sy69 both in vitro and in planta. The results provide support for the use of the quorum‐quenching functionality of B. simplex in the integrated control of the devastating fire blight pathogen.     

A novel enhanced dot blot immunoassay using colorimetric biosensor for detection of Toxoplasma gondii infection

This study reports development of a novel point of care assay, namely an enhanced immuno-dot blot assay, for discrimination of anti-Toxoplasma IgG and anti-Toxoplasma IgM antibodies. This method has been designed based on formation of a sandwich complex between a gold nanoprobe (chitosan gold nanoparticle-anti-human IgG or anti-IgM) and anti- Toxoplasma lysate antigen (TLA) which holds anti-TLA antibodies, either IgG or IgM. Briefly, anti-human IgG or anti-IgM antibody was conjugated to chitosan gold nanoparticles via glutaraldehyde chemistry. Then, lysate antigen was immobilized on the surface of nitrocellulose membrane, which followed by addition of the sera sample and gold nanoprobes. The positive signals were readily detectable via observation with naked eye. This positive color change was further intensified via gold enhancement chemistry. The intensity of biosensor signal was proportional to the concentration of active antibodies on the surface of nanoparticles, titer of T. gondii antibodies in the sera samples and concentration of Toxoplasma lysate antigen coated on the nitrocellulose membrane. A minimum concentration to use the antibodies for conjugation, to detect titer of Toxoplasma IgG and IgM antibodies, and the concentration of TLA coated in nitrocellulose membrane were 0.5 mg/mL, 2 IU/mL, 10 IU/mL, and 20 μg/mL, respectively. This enhanced immuno-dot blot assay offers a simple diagnostic technique without expensive equipment requirement for distinguishing of anti- T. gondii IgM and IgG antibodies in field conditions, pregnant women, and immunocompromised patients.     

检测有机磷农药残留生物传感器的温度特性

为了考察检测有机磷农药残留用量热式生物传感器的温度特性,本文采用分光光度法研究了固定化鸡肝酯酶的活力、农药敌敌畏对鸡肝酯酶的抑制等与温度的关系。在生物传感器的实测条件下,研究了不同温度固定化酶活力的操作流失情况。研究显示,温度对酶反应的影响很大,控温精度的提高将增强检测的精度和稳定性。温度55～60℃时,是酶反应速度最快的区域;抑制时间5 min时,农药敌敌畏对鸡肝脂酶产生了明显的抑制作用。在农药浓度小于1 mg/L的区域,随浓度变化的相对抑制关系接近于线性;一定浓度的农药敌敌畏对酶的抑制程度随着温度的变     

生物传感器在水产品新鲜度检测中的应用与研究进展

新鲜度是判断水产品品质的一项重要指标。传统新鲜度检测方法大多繁琐、费时费力,建立快速便捷的检测方法具有重要的现实意义和学术价值。生物传感器操作简单、检测快速、成本低,近年来被广泛研究应用于水产品新鲜度的检测。本文在简要阐述传统和新型新鲜度检测方法的基础上,综述了生物传感器在水产品新鲜度检测上的应用,着重介绍了酶固定载体和纳米传感器方面的最新进展,并概述用于水产品新鲜度检测的生物传感器技术当前存在的问题及发展趋势,旨在为该光传感器的后续研发提供参考。     

YB菌株生化需氧量的测定及对河豚毒素毒性的响应

[目的]了解自有YB菌株在BOD生物传感器及河豚毒素毒性传感器中的作用。[方法]以YB菌株作为响应菌株,采用夹层膜法制作固定化微生物膜,组装成微生物传感器并测试其性能。[结果]YB菌株在33℃,pH为7.2的条件下对BOD响应良好,在此条件下静态法测得传感器标准曲线为y=0.061 2x-0.001 3,线性相关系数为0.995;动态法测得传感器标准曲线为y=0.072 2x-0.031 9,线性相关系数为0.996。同时,YB菌株对河豚毒素基本没有响应。[结论]自有YB菌株可以作为BOD微生物传感器的敏感菌,但不能作为河豚毒素毒性传感器测试菌株。     

基于DNA双链电荷转移的Hg2+电化学生物传感器的研究

基于DNA双链电荷转移原理,利用胸腺嘧啶(thymine)与Hg2的特异性识别和计时电量法构建了一种高灵敏检测水溶液中Hg2的电化学生物传感器.该传感器将含有1个T-T碱基错配对的DNA互补双链通过Au-S键自组装在金电极表面,运用计时电量法在含有亚甲基蓝的铁氰化钾溶液中进行测定.T-T错配阻断了DNA双链内部电荷转移,而Hg2通过T-Hg2-T配位作用与双链DNA特异性结合并形成DNA双链内部电荷转移通路,引起电极表面计时电量的变化.计时电量测定结果显示:在亚甲基蓝的还原峰电位(-380mV)附近,计时电量随着溶液中的Hg2+浓度的增大而增加,Hg2+浓度在1.0 nmol/L～ 104 nmol/L范围内,计时电量的变化量与Hg2浓度的对数呈良好的线性关系,线性相关系数(R2)为0.997,检测限为0.5 nmol/L(S/N=3).干扰实验表明,该传感器对Hg2具有良好的特异性和选择性.