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Title: 利用功能化金奈米粒子於快速基因檢測平台開發
Development of a fast analysis platform for detecting specific gene using functional gold nanoparticles
Authors: Li, Han-Wei
Contributors: 國立臺灣海洋大學:生命科學暨生物科技學系
Keywords: 基因檢測;金奈米粒子;聚腺嘌呤;聚合酶連鎖反應;臨床診斷
gene detection;gold nanoparticles;poly-adenine;polymerase chain reaction;clinical diagnosis
Date: 2017
Issue Date: 2018-08-22T06:31:27Z
Abstract: 基因檢測於臨床醫學上能提供相當重要的指標,目前主要的檢測方法為利用即時定量聚合酶連鎖反應儀 (Real time Quantitative Polymerase Chain Reaction Machine) 進行試驗分析,雖然具有高靈敏度但耗時且成本較高,未能滿足高效且低成本的臨床檢測需求。因此本計劃主要目的是開發功能性核酸修飾之金奈米粒子,利用金奈米粒子與腺嘌呤有高度親和力的特性,以尾端為聚腺嘌呤之核酸修飾於金奈米粒子,相較於傳統使用硫醇基修飾核酸於金奈米粒子,成本約可節省五倍以上。結合聚合酶連鎖反應儀擴增目標基因片段後,藉由修飾有核酸之金奈米粒子能專一性與目標基因互補雜交而造成金奈米粒子聚集的原理,可於可見光波段 (~520 nm) 檢測目標基因。此方法提供一個快速基因檢測平台,能短時間進行基因檢測,並具定量功能,相較於傳統檢測法能更貼近臨床醫學上之簡單、快速、低成本、高靈敏度和專一性的需求,並期望此研究開發快速基因檢測平台可應用於病毒檢測、甲基化基因分析或其它癌症基因分析。
Gene detection is the one of the important diagnostic tests that can provide useful information to understand the fitness or illness of organs and systems, which enables early detection of serious diseases. The current method for analysis of DNA is through amplification and quantification of DNA by quantitative polymerase chain reaction (qPCR), which has high sensitivity and accuracy. Although qPCR method has these advantages, the high cost of qPCR instrument limits its wide application. In this project, we aim at developing a highly sensitive, high throughput and low cost analysis platform for specific DNA detection by employing noncovalent DNA-modified-gold nanoparticles as probes (probe DNAAu NPs). Considering the disadvantage of the high cost of using thiolated DNA to modify on Au NPs surface, we will use DNA containing poly-adenine (poly-A) tail having high affinity towards Au NPs to prepare probe DNAAu NPs. We believe that employing poly-A tailed DNA can reduce the cost to 20% in comparison with using thiolated DNA probe. The simple and rapid colorimetric assay will be achieved by target DNA-induced aggregation of probe DNAAu NPs when the target DNA is hybridized with probe DNA. As a result, the concentration can be determined by monitoring the absorption at ~520 nm. This method can provide a rapid, sensitive, highly specific and high throughput analysis platform for detecting DNA. These advantages meet the requirements for the clinical diagnosis, and broader application than qPCR method becauseof their low cost and easy handling of instruments. We believe this analysis platform will become a potential tool for DNA detection in commercial clinical diagnosis.
URI: http://ethesys.lib.ntou.edu.tw/cgi-bin/gs32/gsweb.cgi?o=dstdcdr&s=G001043B001.id
Appears in Collections:[生命科學暨生物科技學系] 博碩士論文

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