陳力騏Chen, Richie L.C.臺灣大學:生物產業機電工程學研究所聶其華Nieh, Chi-HuaChi-HuaNieh2010-05-052018-07-102010-05-052018-07-102009U0001-0707200911211100http://ntur.lib.ntu.edu.tw//handle/246246/180246本研究開發兩種拋棄式檢測試片應用於茶品與蝴蝶蘭的品質管理。茶品品管檢測試片係利用拋棄式網印碳電極搭配流動注射系統檢測茶湯的氧化還原電位，藉此評定茶品發酵度。此法可減少電極表面上多元酚類的吸附(CV<1.2%)且達到高通量檢測 (樣品檢測數量 >20 hr-1)。透過檢測不同發酵度之自製包種茶，顯示此法與高效能液相層析法呈現高度相關性。研究另一檢測試片為蘭花蕙蘭嵌紋病毒(CymMV)檢測用免疫層析試片。將多株抗體結合40nm金粒子，可利用肉眼觀察到抗原抗體結合之呈色變化。本研究中，利用肌紅蛋白固定於金奈米粒子，透過肌紅蛋白內之heme基來增強luminol- H2O2 之化學發光強度，藉此降低試片檢測極限。Two types of disposable sensing strips were developed for the quality control of two economic agricultural products, tea and moth orchids. The sensing strip for tea quality is a screen-printed carbon-paste disposable electrode that can be incorporated into a flow injection manifold to estimate tea fermentation degrees based on the redox potentials of tea infusions. The flow-injection analytical manner reduced the adsorption of polyphenols on the electrode surface, reproducible (CV<1.2%) and high-throughput analytical results (>20 hr-1) were obtained. For pouchong tea samples with different fermentation degrees, the data show a high correlation with those obtained by HPLC.he second one is an immunochromatographic strip for detecting the Cymbidium mosaic virus in moth orchids (phalaenopsis spp.). To visualize the immunochromatographic process, polyclonal antibodies against the coat protein of the virus were attached onto 40nm gold nanoparticles. In the present study, myoglobin was immobilized onto the gold nanoparticles to enhance the luminol-H2O2 chemiluminescence by the prosthetic heme group of myoglobin.誌謝 i文摘要 iibstract iiiable of Contents ivist of Figures viiist of Tables ixomenclature xhapter 1 Introduction 1hapter 2 Literature Review 3.1 Estimation of tea fermentation degree 3.1.1 Chromatographic determination of tea catechins 3.1.2 CE determination of tea catechins 8.1.3 Spectrographic methods 9.1.4 Electrochemical and other methods 9.1.5 Fouling problems on the electrode surface 11.2 Virus diagnosis in orchids 12.2.1 Immunoassay procedures 12.2.1.1 Agar gel double diffusion method 12.2.1.2 Enzyme-linked immunosorbent assay (ELISA) 12.2.1.3 Quartz crystal microbalance (QCM) immunosensors 15.2.1.4 Magnetoreduction assay (MRA) 15.2.2 Nucleic acids procedures 17.2.2.1 Polymerase chain reaction (PCR)-based methods 17.2.2.2 QCM-based DNA biosensors 20.2.3 Other analytical methods 20.2.4 Novel immunochromatographic strip for biodiagnosis 21.2.5 Properties and applications of AuNPs in biodiagnosis 23hapter 3 Materials and Methods 26.1 Estimation of tea fermentation degree 26.1.1 Chemicals 26.1.2 Preparation of buffers and reagents 26.1.3 Preparation of tea infusions 26.1.4 Flow-injection manifold 27.1.5 Working electrodes 29.1.6 Manufacture of tea samples 29.1.7 HPLC estimation of tea catehins 31.2 Detection of CymMV and ORSV by a immunochromatographic strip 32.2.1 Chemicals 32.2.2 Preparation of reagents 32.2.3 Preparation of gold colloid 33.2.4 Conjugation of catalyst-AuNPs 34.2.4.1 Preparation of HRP-AuNPs 34.2.4.2 Preparation of Mb-AuNPs 34.2.4.3 Preparation of Mb-S-AuNPs 34.2.5 CL measurement of Mb-AuNP conjugates 37.2.6 Preparation of anti-CymMV IgG&Mb-AuNP conjugates 37.2.7 Immunochromatographic strip test 37hapter 4 Results and Discussion 39.1 Estimation of tea fermentation degree 39.1.1 Reliability of FIA system 39.1.2 Comparison of different working electrodes on sensor response 40.1.3 Redox potential measurement of black tea/green tea mixtures 41.1.4 Effect of indoor withering on redox potential and depletion of tea catechins 45.1.5 Measurements of commercial tea 49.2 Detection of CymMV and ORSV by an immunochromatographic strip 50.2.1 Synthesis of AuNPs 50.2.2 Preliminary test of catalyst-immobilized AuNPs 53.2.3 Properties of Mb-AuNP conjugates 55.2.4 Optimization of reagents in CL system 57.2.5 Effect of Mb-AuNP conjugates on CL intensity 60.2.6 Preliminary test of immunochromatographic strip for CymMV detection 61hapter 5 Conclusions 62eferences 64application/pdf664024 bytesapplication/pdfen-US電位法流動注射系統茶品發酵度免疫層析試片蘭花病毒化學發光Potentiometric flow injectionTea fermentation degreeImmunochromatographic stripOrchid virusChemiluminescencethesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/180246/1/ntu-98-R96631005-1.pdf