氟罗沙星表面等离子共振生物传感器的开发研究

研究利用表面等离子共振生物传感器（SPR）检测氟罗沙星标准品的检测条件,绘制标准曲线,为实际检测提供依据。试验中采用氨基偶联法对芯片（SM5）活化及固定氟罗沙星-OVA偶联物,对抗原在芯片表面的固定条件及抗体浓度的选择进行优化。结果表明,芯片活化及抗原固定时,选用pH 4.5的醋酸缓冲液及50 mmol/L氟罗沙星-OVA偶联物可以得到较理想的固定响应值;试验中绘制标准曲线的最低检测限为1 ng/mL,检测范围为0～20 ng/mL。抗体1∶400稀释时可达到较低的检测限,为实际样品检测提供检测依据。     

黄嘌呤氧化酶传感器的研制及其对鱼鲜度的检测

将黄嘌呤氧化酶以共价连接法固定于二醋酸纤维素薄膜上，制成的酶膜与极谱式氧电极共同组成对次黄嘌呤进行定量测定的黄嘌呤氧化酶传感器。该传感器测定范围为１０～１２０ｍｇ／Ｌ，响应时间１０ｓ，不同浓度次黄嘌呤标准溶液对传感器１０ｓ响应值的相关系数为γ＝－０．９９６６，次黄嘌呤标准溶液重复测定６０次（１０ｓ响应值）的变异系数为ＣＶ＝０．３０３％。应用该传感器与分光光度法对鱼体次黄嘌呤含量进行了测定比较，二者具有良好的相关性。     

表面等离子共振生物传感器检测培氟沙星的方法研究

研究表面等离子共振生物传感器检测缓冲液中培氟沙星的方法,为样品的检测提供依据。采用抑制法,将培氟沙星与卵清蛋白的偶联物固定在芯片上,将抗体与不同浓度的培氟沙星混合后通入芯片表面,优化反应条件,构建标准曲线。结果显示,选择1∶1000稀释的抗体,流速7 μL/min,反应时间4 min,构建标准曲线,可以达到较低的检测限。利用SPR生物传感器建立快速检测缓冲液中培氟沙星的方法,检测限为0.8 ng/mL,检测时间为4 min,为样品检测奠定基础。     

农药残留快速检测技术的最新进展

酶抑制法、免疫分析法和生物传感器在农药残留快速检测中发挥着越来越重要的作用,日益受到国际社会的高度重视。本研究详细介绍了酶抑制法、免疫分析法和生物传感器这3种方法的检测原理、研究现状及实际应用情况,并对农药残留快速检测技术的未来发展趋势进行了展望。     

The Study and Development of Environment-Monitoring Biosensors

A review on environment-monitoring biosensors is presented.During recent years,various environment-monitoring biosensors are fast developed.They are applied to various areas,such as aquatic environment,atmosphere environment et al.The developing trend of environment_monitoring biosensors is also discussed.     

A Simple and Reliable Assay for Detecting Specific Nucleotide Sequences in Plants Using Optical Thin-film Biosensor Chips

Here we report the adaptation and optimization of an efficient, accurate and inexpensive assay that employs custom-designed silicon-based optical thin-film biosensor chips to detect unique transgenes in genetically modified （GM） crops and SN-P markers in model plant genomes. Briefly, aldehyde-attached sequence-specific singlestranded oligonucleotide probes are arrayed and covalently attached to a hydrazine-derivatized biosensor chip surface. Unique DNA sequences （or genes） are detected by hybridizing biotinylated PCR amplicons of the DNA sequences to probes on the chip surface. In the SN-P assay, target sequences （PCR amplicons） are hybridized in the presence of a mixture of biotinylated detector probes and a thermostable DNA ligase. Only perfect matches between the probe and target sequences, but not those with even a single nucleotide mismatch, can be covalently fixed on the chip surface. In both cases, the presence of specific target sequences is siL, nified by a color change on the chip surface （gold to blue/purple） after brief incubation with an anti-biotin IgG horseradish peroxidase （HRP） to generate a precipitable product from an HRP substrate.     

口蹄疫及检测技术进展

口蹄疫包括七个主要的血清型,被国际兽疫局定为一类传染病,我国农业部也将其列为一类传染病。因为近年频繁爆发,严重威胁到畜禽生产和人类自身生存安全,同时由于亚型多、抗原变异性强、宿主范围广等原因,其生态学和公共卫生学意义有极其重要的作用,本文就它的感染机制和各种诊断与防制措施进行概述,从而阐述目前口蹄疫的诊断技术进展状况和进展。     

N-酰基-高丝氨酸内酯类群体感应信号分子检测方法的建立

N-酰基-高丝氨酸内酯（AHL）类化合物是革兰氏阴性菌群体感应系统中最重要的一类信号分子，并调控许多生理特性的表达。但由于AHLs分子为小分子物质，且AHL产生菌所产生的AHL的浓度极低，因此建立有效检测AHLs方法具有非常重要的意义。本研究利用两种细菌生物感应器Chromobacterium violaceum CVO26和Agrobacterium tumefaciens A136（pCF218/pCF372）建立了琼脂扩散法、薄层层析与细菌生物感应器相结合（TLC-Biosensor）、β-半乳糖苷酶活法等检测AHL的生物学方法,为研究革兰氏阴性菌的群体感应系统提供了有效手段。     

生物传感器及其发展概况

简要介绍了生物传感器的概念、结构、基本原理和国内外发展概况。     

Optimization of an oxygen-based approach for community-level physiological profiling of soils

Current approaches for rapid assessment of carbon source utilization by whole soil communities (i.e., community-level physiological profiling or CLPP) provide a limited, biased view of microbial communities with little connection to in situ activities. We developed an alternative CLPP approach based upon fluorometric detection of dissolved oxygen consumption in a microtiter platform which offers flexible manipulation of experimental factors. In the attempt to reduce oxygen re-dissolution, the wells were filled with liquid to very near the top and sealed. We found that filling the wells with 240 vs. 150 μl of sample improved the sensitivity of the system to discern both the response to a substrate amendment as low as 10 mg l⁻¹ and un-amended, endogenous respiration. The preparation of a soil slurry facilitates inoculation into the microplate. Disruption of soil samples had a limited effect on the endogenous respiration in comparison to intact soil microbags in a 24-well microplate. Storage time (up to 33 days) reduced the level of activity in intact soil microbags but not in disrupted samples. A microcosm fertilization experiment was set to study the effects of N availability on the respiratory response in the plates. The use of soil organic carbon (SOC) and amended C-substrates (50 mg l⁻¹) was increased by the addition of nitrogen (N) in the plate, and appeared N-limited shortly after microcosm fertilization. The addition of the eukaryotic inhibitor cycloheximide delayed the initial increase in fluorescence (time to minimum response) of several C sources (casein, acetate, asparagine, coumaric acid), varying among soils, which could be explained by the fungal use of these compounds. However, the extent of the inhibition caused by cycloheximide did not increase at higher fungal to bacteria ratios as estimated by PLFA analysis, indicating that the direct estimation of the fungal biomass from cycloheximide addition is not feasible. This paper provides an optimized, standardized protocol for soil analysis, and sets the basis for further validation studies that will continue to define the underlying capabilities/biases of this approach.