本文選題：禽白血病病毒 + 免疫傳感器�。� 參考：《山東農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：近幾年來(lái),由J亞群禽白血病病毒(ALVs-J)引起的家禽死亡及淘汰率逐年上升,造成的經(jīng)濟(jì)損失逐年增加,也嚴(yán)重危害了我們的食品安全。目前檢測(cè)ALVs-J的方法多存在耗時(shí)長(zhǎng)、成本高等缺點(diǎn)。因此我們將研究重點(diǎn)放在了低消耗、高靈敏、響應(yīng)快速、設(shè)備低廉、操作簡(jiǎn)便的免疫分析法上。本文研究了摻氮多孔碳材料、類石墨相氮化碳、納米纖維素等納米碳材料及其復(fù)合材料的性能,構(gòu)建了幾種不同類型的免疫傳感器,實(shí)現(xiàn)了對(duì)ALVs-J的超靈敏檢測(cè)。主要分為以下四部分:(1)以納米金(AuNPs)、伴刀豆球蛋白A(Con A)作為基底,鉑納米粒子修飾的摻氮多孔碳材料(Pt/PCN)做為探針構(gòu)建了一種新型的電化學(xué)免疫傳感器。根據(jù)之前的報(bào)道,Pt/PCN比單純的Pt或者PCN有更高的催化活性。利用Pt/PCN可大大提高免疫傳感器的靈敏度。Pt/PCN的類過(guò)氧化物酶活性可以催化對(duì)苯二酚的氧化,根據(jù)生成的對(duì)苯醌的電化學(xué)活性,可以實(shí)現(xiàn)對(duì)ALVs-J的檢測(cè)。在最佳實(shí)驗(yàn)條件下,電化學(xué)免疫傳感器所得到的還原峰電流與ALVs-J濃度的對(duì)數(shù)值在102.02 to 104.30 TCID50/m L范圍內(nèi)成正比,檢測(cè)限為107 TCID50/mL(S/N=3)。此電化學(xué)免疫傳感器還展現(xiàn)出良好的選擇性、重現(xiàn)性和穩(wěn)定性。(2)本研究基于納米金與類石墨相氮化碳的復(fù)合材料(Au NPs/g-C3N4)構(gòu)建了一種新型三明治結(jié)構(gòu)電化學(xué)免疫傳感器,實(shí)現(xiàn)了對(duì)ALVs-J的超靈敏檢測(cè)。該免疫傳感器具有三重信號(hào)放大策略,提高了檢測(cè)的靈敏性。以Au NPs/g-C3N4為電極修飾材料,增加了電極表面積從而增加了捕獲一抗的量;納米金具有類氧化物酶性質(zhì),可以催化檢測(cè)液中的葡萄糖生成過(guò)氧化氫和葡萄糖酸;生成過(guò)氧化氫后,所負(fù)載的辣根過(guò)氧化物酶(HRP)對(duì)電信號(hào)物質(zhì)甲苯胺藍(lán)(Tb)顯示出極大的電化學(xué)生物催化活性。通過(guò)對(duì)比不同的傳感器基底和探針,證實(shí)了傳感器的信號(hào)放大作用。在最佳實(shí)驗(yàn)條件下,電化學(xué)免疫傳感器所得到的氧化峰電流與ALVs-J濃度的對(duì)數(shù)值在102.22 to 104.30 TCID50/mL范圍內(nèi)成正比,檢測(cè)限為122 TCID50/mL(S/N=3)。此電化學(xué)免疫傳感器還展現(xiàn)出良好的選擇性、重現(xiàn)性和穩(wěn)定性。(3)本研究以石墨烯為基底、納米金納米纖維素復(fù)合物為探針構(gòu)建了一種新型電化學(xué)免疫傳感器,實(shí)現(xiàn)了對(duì)禽白血病病毒的高靈敏檢測(cè)。石墨烯提高了甲苯胺藍(lán)的負(fù)載量,也增加了捕獲一抗的量;在過(guò)氧化氫存在的條件下,所負(fù)載的辣根過(guò)氧化物酶(HRP)對(duì)電信號(hào)物質(zhì)甲苯胺藍(lán)(Tb)顯示出極大的電化學(xué)生物催化活性。通過(guò)對(duì)比不同的傳感器探針,證實(shí)了傳感器的信號(hào)放大作用。在最佳實(shí)驗(yàn)條件下,電化學(xué)免疫傳感器所得到的還原峰電流與ALVs-J濃度的對(duì)數(shù)值在102.08 to 104.00 TCID50/mL范圍內(nèi)成正比,檢測(cè)限為116 TCID50/mL(S/N=3)。此電化學(xué)免疫傳感器還展現(xiàn)出良好的選擇性、重現(xiàn)性和穩(wěn)定性。(4)利用自增強(qiáng)電化學(xué)發(fā)光,以修飾Ru(Ⅱ)復(fù)合物的納米纖維素為探針構(gòu)建了一種新型免疫傳感器,實(shí)現(xiàn)了ALVs-J的超靈敏檢測(cè)。通過(guò)共反應(yīng)劑樹狀高分子(PAMAM)與發(fā)光物質(zhì)二(2,2’-聯(lián)吡啶)-4’-甲基-4-羧基聯(lián)吡啶-釕N-琥珀酰亞胺酯-二(六氟磷酸鹽)的復(fù)合合成了理想的發(fā)光基團(tuán)。與普通的分子間ECL反應(yīng)相比,這種分子內(nèi)的反應(yīng)電子轉(zhuǎn)移路徑短、能量損失少,所以表現(xiàn)出了更高的發(fā)光效率。而且,應(yīng)用Au-PAMAM可以提高Ab1和Ru的負(fù)載量。在最佳實(shí)驗(yàn)條件下,電化學(xué)發(fā)光免疫傳感器所得到的ECL強(qiáng)度與ALVs-J濃度的對(duì)數(shù)值在102.02 to 104.30 TCID50/mL范圍內(nèi)成正比,檢測(cè)限為101TCID50/mL(S/N=3)。此電化學(xué)免疫傳感器還展現(xiàn)出良好的選擇性、重現(xiàn)性和穩(wěn)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.
【學(xué)位授予單位】：山東農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O657.1

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