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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 謝學真(Hsyue-Jen Hsieh) | |
dc.contributor.author | Cheng-Han Tsai | en |
dc.contributor.author | 蔡承翰 | zh_TW |
dc.date.accessioned | 2021-06-16T23:52:46Z | - |
dc.date.available | 2016-07-30 | |
dc.date.copyright | 2012-07-20 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-07-20 | |
dc.identifier.citation | [1] Guzman MG. Dengue, an update. Trop Med Int Health. 2007;12:5.
[2] Dengue and dengue hemorrhagic fever among adults: Clinical outcomes related to viremia, serotypes, and antibody response. J Infect Dis. 2008;197:817-24. [3] Alcon S, Talarmin A, Debruyne M, Falconar A, Deubel V, Flamand M. Enzyme-linked immunosorbent assay specific to dengue virus type 1 nonstructural protein NS1 reveals circulation of the antigen in the blood during the experiencing primary acute phase of disease in patients or secondary infections. J Clin Microbiol. 2002;40:376-81. [4] W. H. O. Dengue, countries or areas at risk, 2010. [http://www. who. int/topics/dengue/en/]2010. [5] Dussart P, Labeau B, Lagathu G, Louis P, Nunes MRT, Rodrigues SG, et al. Evaluation of an enzyme immunoassay for detection of dengue virus NS1 antigen in human serum. Clin Vaccine Immunol. 2006;13:1185-9. [6] Young PR, Hilditch PA, Bletchly C, Halloran W. An antigen capture enzyme-linked immunosorbent assay reveals high levels of the dengue virus protein NS1 in the sera of infected patients. J Clin Microbiol. 2000;38:1053-7. [7] Tomlinson SM, Malmstrom RD, Watowich SJ. New approaches to structure-based discovery of dengue protease inhibitors. Infectious disorders drug targets. 2009;9:327-43. [8] Li PC, Liao MY, Cheng PC, Liang JJ, Liu IJ, Chiu CY, et al. Development of a Humanized Antibody with High Therapeutic Potential against Dengue Virus Type 2. PLoS Negl Trop Dis. 2012;6:e1636. [9] Modis Y, Ogata S, Clements D, Harrison SC. A ligand-binding pocket in the dengue virus envelope glycoprotein. Proceedings of the National Academy of Sciences of the United States of America. 2003;100:6986-91. [10] Lindenbach BD, Rice CM. trans-Complementation of yellow fever virus NS1 reveals a role in early RNA replication. J Virol. 1997;71:9608-17. [11] Mackenzie JM, Jones MK, Young YR. Immunolocalization of the dengue virus nonstructural glycoprotein NS1 suggests a role in viral RNA replication. Virology. 1996;220:232-40. [12] Su CC, Wu TZ, Chen LK, Yang HH, Tai DF. Development of immunochips for the detection of dengue viral antigens. Anal Chim Acta. 2003;479:117-23. [13] Guzman MG, Halstead SB, Artsob H, Buchy P, Farrar J, Gubler DJ, et al. Dengue: a continuing global threat. Nature reviews Microbiology. 2010;8:S7-16. [14] Vaughn DW, Green S, Kalayanarooj S, Innis BL, Nimmannitya S, Suntayakorn S, et al. Dengue in the early febrile phase: Viremia and antibody responses. J Infect Dis. 1997;176:322-30. [15] Wikipedia. Dengue fever. http://en. wikipedia. org/wiki/Dengue_fever. [16] Sia SK, Whitesides GM. Microfluidic devices fabricated in poly(dimethylsiloxane) for biological studies. Electrophoresis. 2003;24:3563-76. [17] Chen CS, Breslauer DN, Luna JI, Grimes A, Chin WC, Leeb LP, et al. Shrinky-Dink microfluidics: 3D polystyrene chips. Lab Chip. 2008;8:622-4. [18] Delamarche E, Juncker D, Schmid H. Microfluidics for processing surfaces and miniaturizing biological assays. Adv Mater. 2005;17:2911-33. [19] Olkkonen J, Lehtinen K, Erho T. Flexographically printed fluidic structures in paper. Anal Chem. 2010;82:10246-50. [20] Martinez AW, Phillips ST, Butte MJ, Whitesides GM. Patterned paper as a platform for inexpensive, low-volume, portable bioassays. Angew Chem Int Edit. 2007;46:1318-20. [21] Cheng CM, Martinez AW, Gong JL, Mace CR, Phillips ST, Carrilho E, et al. Paper-Based ELISA. Angew Chem Int Edit. 2010;49:4771-4. [22] Martinez AW, Phillips ST, Whitesides GM, Carrilho E. Diagnostics for the developing world: microfluidic paper-based analytical devices. Anal Chem. 2010;82:3-10. [23] Li X, Tian JF, Garnier G, Shen W. Fabrication of paper-based microfluidic sensors by printing. Colloid Surface B. 2010;76:564-70. [24] Carrilho E, Martinez AW, Whitesides GM. Understanding Wax Printing: A Simple Micropatterning Process for Paper-Based Microfluidics. Anal Chem. 2009;81:7091-5. [25] Carrilho E, Martinez AW, Whitesides GM. Understanding wax printing: a simple micropatterning process for paper-based microfluidics. Anal Chem. 2009;81:7091-5. [26] Martinez AW, Phillips ST, Whitesides GM. Three-dimensional microfluidic devices fabricated in layered paper and tape. Proceedings of the National Academy of Sciences of the United States of America. 2008;105:19606-11. [27] Lai CY, Tsai WY, Lin SR, Kao CL, Hu HP, King CC, et al. Antibodies to envelope glycoprotein of dengue virus during the natural course of infection are predominantly cross-reactive and recognize epitopes containing highly conserved residues at the fusion loop of domain II. J Virol. 2008;82:6631-43. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/65595 | - |
dc.description.abstract | 本研究著重於發展以紙為基材的檢測系統,用來偵測登革病毒抗原(dengue virus antigen)。以紙為基材的微流體系統相較於傳統的微流體元件,有低元件製造成本以及方便操作等多項優勢,本研究將利用此優勢,使用噴蠟列印法(Wax printing method)製備酵素連結免疫吸附分析法(Enzyme-linked immunosorbent assay)所使用的96孔盤,並且在其上建立一個用來檢測第二型登革病毒(dengue virus type 2, DENV-2)抗原的檢測平台。本研究中所使用的登革病毒抗原分別為第二型登革病毒的第一型非結構蛋白(dengue virus type 2 nonstructural protein 1, NS-1)以及第二型登革病毒的外鞘蛋白(dengue virus type 2 E protein, E protein),利用這兩種登革病毒特有的生物指標分子以及其抗體,進行以紙為基材的酵素免疫分析法,並且利用家用掃描機來記錄每次酵素呈色後的顏色深淺數值,便可以此數值來判斷樣品中登革病毒抗原的濃度。另一方面,本研究也將此登革病毒抗原以及抗體的組合,應用於以紙為基材的側邊流免疫檢測元件(lateral flow immunoassay device)中,利用有奈米金粒子結合的抗第二型登革病毒第一型非結構蛋白的抗體(anti-DENV-2 NS-1 immunoglobulin),將可以在紙上進行第二型登革病毒抗原的快速篩檢。實驗結果顯示,在紙上進行酵素免疫分析法,在適當的清洗步驟下,能獲得較低的訊號背景值以及較高的檢測靈敏度。本研究中所開發的登革病毒抗原酵素免疫分析檢測系統的靈敏度可以達到1 (ng/ml),並且可以使用肉眼就可以清楚分辨不同濃度的抗原在紙上所顯示的顏色。使用酵素結合之單株抗體(enzyme-conjugated monoclonal antibody)來進行第二型登革病毒外鞘蛋白的檢測,同樣可以獲得可由肉眼分辨的的顏色訊號,使我們可以快速的利用此技術定性上檢測病毒的外鞘蛋白。使用以紙為基材的酵素免疫分析平台,不論在登革病毒抗原的定性或是定量上,皆能有清晰且準確的檢測結果。 | zh_TW |
dc.description.abstract | This research focused on the development of a low-cost, easy-to-handle, and robust approach to detect one of the deadly diseases in tropical medicine—dengue. To achieve this goal, we have developed a paper-based ELISA (P-ELISA) —a new, inexpensive, and easy-to-handle platform for biochemical analysis—to detect the dengue virus type 2 nonstructural protein 1 (DENV-2 NS-1) and E protein.
We introduced a new paper fabrication method, called wax printing method, to fabricate the paper in a rapid and easy way. The wax printed on the paper will form a hydrophilic-hydrophobic boundary after reflowing the wax by heating. We use this wax printing technique to fabricate a 96-well plate on the filter paper to prepare a paper-based platform to do ELISA for detecting dengue antigen proteins. The current results indicated that paper-based sandwich ELISA was capable of detecting ~1 (ng/ml) of DENV-2 NS-1. Also, indirect ELISA for dengue virus type 2 envelope protein (DENV-2 E protein) gives a distinct color intensity even at one-hundred folds of VLP (virus-like particle solution) dilution. These promising results, we believe, have a wide range of potential applications such as the detections of various proteins or antigens associated with many tropical infectious diseases. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T23:52:46Z (GMT). No. of bitstreams: 1 ntu-101-R99524026-1.pdf: 1762166 bytes, checksum: 802ebb2c8dc4804c1cf67b1bd063b7cb (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | Contents
誌謝 I 中文摘要 III Abstract V Contents VII List of Figures XI List of Tables XIII 縮寫對照 XV Chapter 1 Introduction 1 1. 1 Epidemic of dengue 1 1.2 Dengue virus antigen proteins 2 1. 3 Methodology for dengue diagnosis 5 1. 4 Application of microfluidic device 6 1. 5 Fabrication of paper-based microfluidic device 7 1. 5. 1 Photolithography 8 1. 5. 2 Chemical modification by alkyl ketene dimer 9 1. 5. 3 Wax printing method 10 1. 6 Overview of the research 12 Chapter 2 Material and Methods 13 2. 1 Preparation of a P-ELISA plate 13 2. 2 P-ELISA for DENV-2 NS-1 and E protein 13 2. 2. 1 Antibody 13 2. 2. 2 Antigen protein 14 2. 2. 3 Reagent for ELISA 14 2. 3 Building NS-1 calibration curve by using P- ELISA 14 2. 4 Demonstration of P-ELISA for DENV-2 E protein 17 Chapter 3 Results 19 3. 1 P-ELISA plate preparation 19 3. 2 Washing method testing 21 3. 2. 1 Testing of buffer washing strength 21 3. 2. 2 Testing of drop washing 23 3. 2. 3 Test of dip washing 25 3. 3 Testing of primary antibody concentration 27 3. 4 Protocol for P-ELISA to detect DENV-2 NS-1 29 3. 5 Calibration curve of DENV-2 NS-1 31 3. 5. 1 Calibration curve fitting 31 3. 6 Comparison between P-ELISA and conventional ELISA 35 3. 7 Demonstration of P-ELISA for DENV-2 E protein 37 3. 7. 1 Protocol for P-ELISA to detect DENV-2 E protein 37 3. 7. 2 DENV-2 E protein detection 39 Chapter 4 Discussion 41 4. 1 P-ELISA plate preparation 41 4. 2 P-ELISA for DENV-2 NS-1 and E protein 42 4. 2. 1 Building protocol for P-ELISA 42 4. 2. 2 Building DENV-2 NS-1 calibration curve by using P-ELISA 45 4. 3 Demonstration of P-ELISA for DENV-2 E protein 47 Chapter 5 Conclusions 49 5. 1 Conclusions 49 5. 2 Future works 50 References 51 | |
dc.language.iso | en | |
dc.title | 偵測登革病毒抗原之紙基檢測系統之開發 | zh_TW |
dc.title | Paper-based Assay System for Detecting Dengue Viral Antigens | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 鄭兆?(Chao-Min Cheng) | |
dc.contributor.oralexamcommittee | 吳漢忠(Han-Chung Wu),侯劭毅(Shao-Yi Hou) | |
dc.subject.keyword | 噴蠟列印法,登革熱檢測,紙基酵素連結免疫吸附分析法, | zh_TW |
dc.subject.keyword | wax printing method,dengue fever diagnosis,Paper-based ELISA, | en |
dc.relation.page | 54 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2012-07-20 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 化學工程學研究所 | zh_TW |
顯示於系所單位: | 化學工程學系 |
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