纳米碳材料在禽白血病病毒电化学免疫分析中的应用

发布时间：2018-07-01 19:01

本文选题：禽白血病病毒 + 免疫传感器　；参考：《山东农业大学》2017年硕士论文

【摘要】：近几年来,由J亚群禽白血病病毒(ALVs-J)引起的家禽死亡及淘汰率逐年上升,造成的经济损失逐年增加,也严重危害了我们的食品安全。目前检测ALVs-J的方法多存在耗时长、成本高等缺点。因此我们将研究重点放在了低消耗、高灵敏、响应快速、设备低廉、操作简便的免疫分析法上。本文研究了掺氮多孔碳材料、类石墨相氮化碳、纳米纤维素等纳米碳材料及其复合材料的性能,构建了几种不同类型的免疫传感器,实现了对ALVs-J的超灵敏检测。主要分为以下四部分:(1)以纳米金(AuNPs)、伴刀豆球蛋白A(Con A)作为基底,铂纳米粒子修饰的掺氮多孔碳材料(Pt/PCN)做为探针构建了一种新型的电化学免疫传感器。根据之前的报道,Pt/PCN比单纯的Pt或者PCN有更高的催化活性。利用Pt/PCN可大大提高免疫传感器的灵敏度。Pt/PCN的类过氧化物酶活性可以催化对苯二酚的氧化,根据生成的对苯醌的电化学活性,可以实现对ALVs-J的检测。在最佳实验条件下,电化学免疫传感器所得到的还原峰电流与ALVs-J浓度的对数值在102.02 to 104.30 TCID50/m L范围内成正比,检测限为107 TCID50/mL(S/N=3)。此电化学免疫传感器还展现出良好的选择性、重现性和稳定性。(2)本研究基于纳米金与类石墨相氮化碳的复合材料(Au NPs/g-C3N4)构建了一种新型三明治结构电化学免疫传感器,实现了对ALVs-J的超灵敏检测。该免疫传感器具有三重信号放大策略,提高了检测的灵敏性。以Au NPs/g-C3N4为电极修饰材料,增加了电极表面积从而增加了捕获一抗的量;纳米金具有类氧化物酶性质,可以催化检测液中的葡萄糖生成过氧化氢和葡萄糖酸;生成过氧化氢后,所负载的辣根过氧化物酶(HRP)对电信号物质甲苯胺蓝(Tb)显示出极大的电化学生物催化活性。通过对比不同的传感器基底和探针,证实了传感器的信号放大作用。在最佳实验条件下,电化学免疫传感器所得到的氧化峰电流与ALVs-J浓度的对数值在102.22 to 104.30 TCID50/mL范围内成正比,检测限为122 TCID50/mL(S/N=3)。此电化学免疫传感器还展现出良好的选择性、重现性和稳定性。(3)本研究以石墨烯为基底、纳米金纳米纤维素复合物为探针构建了一种新型电化学免疫传感器,实现了对禽白血病病毒的高灵敏检测。石墨烯提高了甲苯胺蓝的负载量,也增加了捕获一抗的量;在过氧化氢存在的条件下,所负载的辣根过氧化物酶(HRP)对电信号物质甲苯胺蓝(Tb)显示出极大的电化学生物催化活性。通过对比不同的传感器探针,证实了传感器的信号放大作用。在最佳实验条件下,电化学免疫传感器所得到的还原峰电流与ALVs-J浓度的对数值在102.08 to 104.00 TCID50/mL范围内成正比,检测限为116 TCID50/mL(S/N=3)。此电化学免疫传感器还展现出良好的选择性、重现性和稳定性。(4)利用自增强电化学发光,以修饰Ru(Ⅱ)复合物的纳米纤维素为探针构建了一种新型免疫传感器,实现了ALVs-J的超灵敏检测。通过共反应剂树状高分子(PAMAM)与发光物质二(2,2’-联吡啶)-4’-甲基-4-羧基联吡啶-钌N-琥珀酰亚胺酯-二(六氟磷酸盐)的复合合成了理想的发光基团。与普通的分子间ECL反应相比,这种分子内的反应电子转移路径短、能量损失少,所以表现出了更高的发光效率。而且,应用Au-PAMAM可以提高Ab1和Ru的负载量。在最佳实验条件下,电化学发光免疫传感器所得到的ECL强度与ALVs-J浓度的对数值在102.02 to 104.30 TCID50/mL范围内成正比,检测限为101TCID50/mL(S/N=3)。此电化学免疫传感器还展现出良好的选择性、重现性和稳定性。
[Abstract]:In recent years, the rate of death and elimination of poultry caused by the J subgroup of avian leukosis virus (ALVs-J) has increased year by year. The economic loss caused by the avian leukemic virus (ALVs-J) has increased year by year, and it also seriously endangers our food safety. At present, the methods of detecting ALVs-J are often time-consuming and costly. Therefore, we focus on low consumption, high sensitivity and quick response. In this paper, the properties of nitrogen doped porous carbon materials, Shi Moxiang carbon nitride and nanoscale carbon nanomaterials and their composites were studied in this paper. Several different types of immunosensor were constructed, and the ultra sensitive detection of ALVs-J was realized. The following four parts were as follows: (1) A UNPs), with concanavin A (Con A) as the base, platinum nanoparticles modified nitrogen doped porous carbon material (Pt/PCN) was used as a probe to construct a new electrochemical immunosensor. According to previous reports, Pt/PCN has a higher catalytic activity than pure Pt or PCN. The use of Pt/PCN can greatly improve the sensitivity of the immunosensor.Pt/PCN. The activity of peroxidase can catalyze the oxidation of hydroquinone. According to the electrochemical activity of the quinone, the ALVs-J can be detected. Under the best experimental conditions, the reduction peak current of the electrochemical immunosensor is proportional to the value of the ALVs-J concentration in the range of 102.02 to 104.30 TCID50/m L, and the detection limit is 107. TCID50/mL (S/N=3). The electrochemical immunosensor also showed good selectivity, reproducibility and stability. (2) a novel sandwich structure electrochemical immunosensor based on nano gold and graphite like carbon nitride (Au NPs/g-C3N4) was developed in this study. The ultra sensitive detection of ALVs-J was realized. The immunosensing apparatus There is a three signal amplification strategy, which improves the sensitivity of detection. Au NPs/g-C3N4 is used as the electrode modifier to increase the surface area of the electrode and increase the amount of the capture one. The nano gold has the properties of the oxide enzyme, which can catalyze the detection of glucose in the liquid to produce hydrogen peroxide and gluconic acid; after the production of hydrogen peroxide, the horseradish horseradish is loaded. The peroxidase (HRP) showed a great electrochemical biocatalytic activity to the electrical signal, toluidine blue (Tb). By comparing the substrates and probes of different sensors, the signal amplification effect was confirmed. Under the optimum experimental conditions, the value of the peak current of the oxidation peak to the concentration of ALVs-J obtained by the electrochemical immunosensor was 102.22 The positive ratio of to 104.30 TCID50/mL was proportional to the detection limit of 122 TCID50/mL (S/N=3). The electrochemical immunosensor also showed good selectivity, reproducibility and stability. (3) a novel electrochemical immunosensor was constructed with graphene as the base and nano gold nano cellulose complex was used as a probe to realize avian leukemia. The high sensitivity test of the virus. Graphene increased the load of toluidine blue and increased the amount of the capture one. Under the presence of hydrogen peroxide, the loaded horseradish peroxidase (HRP) showed great electrochemical bioactivity to the electrical signal, toluidine blue (Tb), and was confirmed by comparison of different sensor probes. Under the best experimental conditions, the reduction peak current obtained by the electrochemical immunosensor is proportional to the value of the ALVs-J concentration in the range of 102.08 to 104 TCID50/mL, and the detection limit is 116 TCID50/mL (S/N=3). The electrochemical immunosensor also shows good selectivity, reproducibility and stability. (4) A new type of immunosensor was constructed with the nano cellulose modified with Ru (II) complex by self enhanced electrochemiluminescence. The super sensitive detection of ALVs-J was realized. Through the co reactants dendrimer (PAMAM) and luminescent substance two (2,2 '- bipyridine) -4' - -4- carboxylic pyridine - ruthenium N- succinimide - two (six fluoro phosphorus) The compound syntheses the ideal luminescent group. Compared with the common intermolecular ECL reaction, the reaction electron transfer path is shorter and the energy loss is less, so it shows higher luminous efficiency. Moreover, the application of Au-PAMAM can increase the load of Ab1 and Ru. Under the best experimental conditions, the electrochemiluminescence immunosensor The obtained ECL strength is proportional to the value of ALVs-J concentration in the range of 102.02 to 104.30 TCID50/mL, and the detection limit is 101TCID50/mL (S/N=3). The electrochemical immunosensor also shows good selectivity, reproducibility and stability.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.1

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