固定化虫荧光素酶光纤传感器

固定化虫荧光素酶光纤传感器蔡谨,王顺光,杨歧生,吉鑫松（浙江大学化工系生化教研室,杭州３１００２７;中国科学院上海生物化学研究所,２０００３１）关键词虫荧光素酶,ＡＴＰ,固定化酶,光纤生物传感器ＡＴＰ是生物体内极为重要的能量物质。如何准确快速地定量Ａ...     

光纤倏逝波生物传感器及其应用

介绍光纤倏逝波生物传感器的基本原理、常用试验方法、基本仪器构建及应用进展。光纤倏逝波生物传感器是基于光波在光纤内以全反射方式传输时产生倏逝波的原理,以生物分子作为敏感元件进行检测的一类新兴传感器。光纤倏逝波生物传感器有望应用于环境监控、食品卫生监控、临床疾病监测、DNA检测和生物战剂检测。     

光纤生物传感器及其在微生物检测中的应用

本文介绍了光纤生物传感器的原理,对光纤传感器制作中的工程学和生物学问题进行了探讨并概述了它的应用情况。     

光纤DNA传感器

光纤ＤＮＡ传感器是ＤＮＡ生物传感器中很有潜力、极具发展前途的一种,具有重要的研究价值和诱人的应用前景。该技术灵敏度高,分析操作简便、快速,为分子生物研究提供了一种新方法,在基础医学、临床医学和预防医学各个领域具有十分广泛的应用前景。本文对光纤ＤＮＡ传感器的原理,ＤＮＡ探针在光纤表而的固定化,光纤ＤＮＡ传感器的类型等方面的研究进展作了评述。     

倏逝波光纤免疫传感器的光纤再生

倏逝波光纤免疫传感器作为一种新兴的检测技术,在环境检测、食品卫生检测及生物医学检测等领域具有广泛的应用前景。为了降低检测成本并达到快速检测,光纤再生问题就显得尤为重要。我们在对倏逝波光纤免疫传感器原理和光纤再生原理介绍的基础上,对光纤再生的酸性碱性溶液、高浓度盐溶液、去污剂等方法进行简要综述,探讨了光纤再生存在的问题,并展望了解决光纤重生后倏逝波光纤免疫传感器的应用前景。     

一种用于盐度测量的光纤传感器

制作了智能水凝胶PDA—C12。根据凝胶的溶胀性质设计制作了一种盐度光纤传感器探头,并实测了在不同盐度溶液中的输出光强,可测范围达到了1．0mol／L以上,为在一般要求下的盐度测量提供了一种新的方法。     

消失波生物传感器及其在DNA与免疫分析中的应用

消失波光纤生物传感器是近年来发展很快的一项的分析技术。它现在已成为分了生物学领域的热门技术。本文叙述消失波生物传感器的识别元件,换能装置以及检测研究系统的研究进展。着重讨论消失波技术在ＤＮＡ检测与免疫检测中的应用。并对这些技术的应用价值做出评价。     

生物传感器快速测定BOD的研究

生化需氧量(biochemicaloxygendemand,BOD)是一种表征水体有机污染程度的综合指标,广泛应用于水体监测和废水处理厂的运行控制。由于BOD的标准测定方法需时5天,不能及时地反映水质状况和反馈处理信息,因此快速测定BOD的方法和仪器化研究近年来得到广泛的重视。利用生物传感器测定BOD是一种有效地快速测定废水中可生化降解有机物的方法。介绍生物传感器的工作原理及其生物敏感材料,讨论BOD传感器的性能参数以及BOD快速测定值(BODst)与标准BOD5值的一致性问题。对现阶段市场上常见的几种BOD快速测定仪进行简单的介绍,并对它们的性能进行比较 。     

压电免疫传感器的表面固定方法

抗原／抗体在石英晶体电极表面固定而不丧失其活性是压电免疫传感器成功的关键,直接影响到它的灵敏度和可重复性等性质。本文分别从固定抗体和抗原的角度介绍了压电免疫传感器表面固定方法。并对戊二醛交联法、自组装单层膜法(SAMs)、蛋白A固定法做了较深入和详细的介绍,展望了其以后的发展前景。     

用光纤生物传感器检测炭疽杆菌、鼠疫杆菌及葡萄球菌肠毒素B

目的:用新研制的光纤生物传感器FOB-3检测多种病原微生物及细菌毒素。方法:利用双抗体夹心法,优化免疫反应的时间和浓度后,用FOB-3分别检测炭疽芽孢及繁殖体、鼠疫F1抗原和葡萄球菌肠毒素B。为便于结果判定,确定截断(cutoff)值,并以Ssignal与Nnoise差值的形式消除荧光信号中的噪声。结果:使用光纤生物传感器FOB-3,在20min内可分别检测到50～1000ng/mL的鼠疫F1抗原、0.1～100μg/mL的葡萄球菌肠毒素B、3×101～3×106CFU/mL的炭疽杆菌繁殖体和4×104～4×107CFU/mL的炭疽芽孢。结论:光纤生物传感器可以灵敏、快速、便捷地检测病原微生物及其毒素。     

Dual-peak long period fiber grating coated with graphene oxide for label-free and specific assays of H5N1 virus

Avian influenza is an acute infectious disease caused by the avian influenza virus (AIV), which has caused enormous economic losses and posed considerable threats to public health. This study aimed to demonstrate an immunosensor based on dispersion turning point long-period fiber grating (DTP-LPFG) integrated with graphene oxide (GO) for the specific detection of a type of AIV H5N1 virus. LPFG was designed to work at DTP, whose dual-peak spacing was very high sensitive to a refractive index. Anti-H5N1 monoclonal antibodies were covalently bonded with the GO film on the fiber surface, thus constructing an immunosensor for the label-free and specific detection of the H5N1 virus. The proposed method was capable of the reliable detection of H5N1 virus with the limit of detection as low as ~1.05 ng/ml within the large range of 1 ng/mL to 25 µg/mL. More importantly, immunoassays of the whole H5N1 virus in clinical samples further confirmed that the GO-integrated DTP-LPFG immunosensor showed very high specificity to the H5N1 virus and demonstrated great potential for clinical use.     

制备肺炎衣原体抗原片检测血清抗体

目的:探索肺炎衣原体抗原片检测血清抗体法在诊断Cpn感染中的实际应用前景。方法:应用进口肺炎衣原体(Cpn)毒株感染Hep-2细胞,分别以瑞氏-姬母萨染色、吖啶橙染色和直接免疫荧光染色等3种方法鉴定Cpn感染细胞。纯化获取大量Cpn抗原,用于制备斑点抗原片。建立微量免疫荧光染色法(MIF)检测血清抗体,诊断Cpn感染。结果:Cpn感染Hep-2细胞的最适条件是用含1μg/mL放线菌酮的维持液,在35℃、5%CO2孵箱中培养7d,并在培养的第0、3、4、5天以2600r/min离心1h,感染成功率极高。染色反应显示,瑞氏-姬母萨染色可将Cpn包涵体染成蓝紫色或红紫色;吖啶橙染色则使Cpn感染的Hep-2细胞呈现鲜明的橘红色;免疫荧光抗体染色后,在Cpn感染细胞内可见亮苹果绿色包涵体。通过斑点抗原荧光抗体染色的方法抽样检测了100份病人血清中的Cpn抗体,其中抗Cpn-IgG抗体的阳性血清共61份,阳性率为61%。与Cpn-外周血单核细胞(Cpn-PBMC)抗原片比较,阳性检出率无明显差别。结论:用Cpn感染细胞制作的Cpn斑点抗原片可用于临床检测血清Cpn-IgG抗体,且具有特异性、敏感性高的特点,但要求检测人员有一定的经验。     

Surface functionalization allowing repetitive use of optical sensors for real‐time detection of antibody‐bacteria interaction

In this study, sensor surface functionalization allowing the repetitive use of a sensing device was evaluated for antibody‐based detection of living bacteria using an optical planar Bragg grating sensor. To achieve regenerable immobilization of bacteria specific antibodies, the heterobifunctional cross‐linker N‐succinimidyl 3‐(2‐pyridyldithio) propionate (SPDP) was linked to an aminosilanized sensor surface and subsequently reduced to expose sulfhydryl groups enabling the covalent conjugation of SPDP‐activated antibodies via disulfide bonds. The immobilization of a capture antibody specific for Staphylococcus aureus on the sensor surface as well as specific binding of S. aureus could be monitored, highlighting the applicability of optical sensors for the specific detection of large biological structures. Reusability of bacteria saturated sensors was successfully demonstrated by cleaving the antibody along with bound bacteria through reduction of disulfide bonds and subsequent re‐functionalization with activated antibody, resulting in comparable sensitivity towards S. aureus.

     

植物长链非编码RNA研究进展

长链非编码RNA(Long non-coding RNA,lncRNA)长度大于200个核苷酸,大量存在于生物体中并具有多种生物学功能。目前,植物中发现的lncRNA大多由RNA聚合酶Ⅱ转录,并通过目标模仿、转录干扰、组蛋白甲基化和DNA甲基化等多种机制介导基因的表达,在植物开花、雄性不育、营养代谢、生物和非生物胁迫等生物过程中起着调节因子的作用。文章综述了近年来发现的植物lncRNA数据库、预测方法、表达及可能的生物学功能。     

Regiospecific orientation of single-chain antibody and atomic force microscope (AFM) images

An antibody containing a genetically engineered lipid group at the N-terminus and a hexahistidinyl tag at the C-terminus (Lpp-scFv-His6) was immobilized in an oriented manner on the surface of liposomes. Liposomes, consisting of antibody and phosphatidylcholine, have been prepared and imaged by AFM. For AFM visualization, the resulting liposomes were bound on the surface of mica by two different mechanisms. The histidine tags present in the antibody molecules of the immunoliposome were anchored to the NiCl₂ treated mica surface. Alternatively, the immunoliposomes were immunochemically bound on antigen-coated mica surface. Both approaches yielded liposomes which were clearly imaged without damage by AFM in ambient condition.     

A novel bioassay for the monitoring of carcinoembryonic antigen in human biofluid using polymeric interface and immunosensing method

Carcinoembryonic antigen (CEA) is a member of a family of cell surface glycoproteins. Recognition of CEA is needed to monitor the physiological status of the patient for treatment and also it is important to assess the severity of the disease. In this work, we reported a novel sandwich‐type electrochemical immunosensor based on gold nanoparticles functionalized cysteamine‐glutaraldehyde (AuNPs‐CysA‐GA) and it successfully designed to detection of the CEA biomarker in a human plasma sample. The AuNPs‐CysA‐GA provides a large surface area for the effective immobilization of CEA antibody, as well as it ascertains the bioactivity and stability of immobilized CEA antigens. Biotinylated‐anti‐CEA antibody (Ab1) was immobilized on the surface of glassy carbon electrode (GCE) modified AuNPs‐CysA‐GA. Also, secondary antibody (HRP‐Ab2) was costed immobilized to complete the sandwich part of immunosensor. Field emission scanning electron microscope (FE‐SEM and EDS), was employed to monitor the sensor fabrication procedure. The immunosensor was used for the detection of CEA using differential pulse voltammetry (DPVs) technique. The proposed interface led to enhancement of accessible surface area for immobilizing high amount of anti‐CEA antibody, increasing electrical conductivity, boosting stability, and biocompatibility. Finally, the low limit of quantitation (LLOQ) of the proposed immunosensor was obtained as 7 ng/mL with the linear range of 0.001‐5 μg/L. The proposed immunoassay was successfully applied for the monitoring of the CEA in unprocessed human plasma samples. Obtained results paved that the proposed bioassay can be used as a novel bioassay for the clinical diagnosis of cancer based on CEA monitoring.     

Novel optical fiber Vernier immunosensor based on cascading Sagnac loops embedded with excessively tilted fiber grating for specific detection of canine distemper virus

A novel optical fiber Vernier effect (VE) biosensor based on cascading Sagnac loops embedded with excessively tilted fiber grating (ExTFG) is proposed for the label free and specific detection of canine distemper virus (CDV). The VE was realized by cascading two different Sagnac loops with similar free spectrum range (FSR), one of which was integrated with panda-type polarization maintaining fiber (PMF) as the reference loop, and the other was embedded with ExTFG as the sensing loop. Owning to the amplified function of the VE, the refractive index (RI) sensitivity of the proposed sensing structure reached −1914.89 nm/RIU, which is approximately 12 times higher than that of the single ExTFG based RI sensor. Furthermore, the ExTFG in sensing loop was modified by graphene oxide (GO) and bio-functionalized by the CDV monoclonal antibodies (anti-CDV MAbs) for the specific detection of the CDV. Experimental results show that the proposed optical fiber Vernier sensor could detect the CDV in buffer solution with concentration as low as 1 pg/mL, and the sensitivity was about −1.18 nm/[log(mg/ml)] in the concentration range of 1 pg/mL ~ 50 ng/mL. The excellent specific and clinical properties of the biosensor were verified by immunoassays for fetal bovine serum, Toxoplasma gondii, rabies virus and CDV serum in sequence. Due to the sensitivity amplification function of VE, dense comb spectrum of the Sagnac loop and the stable interference spectra maintained by the polarized light, the proposed biosensor possesses the combined advantages of high sensitivity, high Q-factor and high stability, which may have potential applications in biosensing fields.     

长链非编码RNA的微RNA海绵作用与呼吸系统疾病

长链非编码RNA（long non-coding RNAs, lncRNAs）是一类由长度大于200个核苷酸组成的长链非编码序列。lncRNAs具有较长的序列,使得lncRNAs具有复杂的二级及三级结构,这也是lncRNAs结合DNA、RNA和蛋白质及其行使复杂功能的结构基础。MicroRNA（miRNAs)是长度在19到25个核苷酸之间的非编码单链RNA分子,是目前研究最多的小分子非编码RNA。而lncRNAs通过结合或者螯合miRNA来调节miRNA丰度,发挥lncRNA的“海绵”作用,从而调控一系列的病理生理过程。lncRNAs及miRNA在呼吸系统疾病的发生、发展、治疗和预后起重要作用。本文就lncRNAs及其“海绵”作用对呼吸系统疾病的影响及可能的机制进行综述。