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利用蛋白質聚合物達到訊號放大 = Signal Amplificatio...

國立高雄大學應用化學系碩士班

利用蛋白質聚合物達到訊號放大 = Signal Amplification by the Generation of Protein Polymer Networks

紀錄類型:	書目-語言資料,印刷品 : 單行本
並列題名:	Signal Amplification by the Generation of Protein Polymer Networks
作者:	林伯烜,
其他團體作者:	國立高雄大學
出版地:	[高雄市]
出版者:	撰者;
出版年:	2012[民101]
面頁冊數:	115面圖，表 : 30公分;
標題:	生物檢測
標題:	streptavidin
電子資源:	http://handle.ncl.edu.tw/11296/ndltd/90904846645347037157
附註:	106年4月25日公開
附註:	含參考書目
摘要註:	現今越來越多新的疾病被發現在世界上，且大多發現在落後的國家。目前雖有適當的技術來檢測疾病，但對資源匱乏的環境而言，檢測仍然是一個挑戰。一個理想的診斷測試其特點應包含廉價、靈敏、特異、操作簡單、快速且穩定（無需冷藏），最重要的是，不需使用昂貴設備。在傳統的細菌檢測技術上，它是耗時且勞動力密集的。例如平板計數法（plate count techniques），酶聯免疫吸附試驗（ELISA），生化試驗和聚合酶鏈反應（PCR）等。這些技術依賴於精密儀器，昂貴試劑和大量的技術工人。對落後的國家和資源貧乏的地區而言，滿足這些需求仍然是一大問題。蛋白質聚集放大法（PCBAM）計畫是一種以視覺檢測和鑑定人類樣本中病原微生物的技術。此新方法的核心在於使用生物素標記牛血清蛋白（bBSA）與鏈黴親和素連結辣根過氧化物酶;（S-HRP）形成類聚合物。該假設是：此類聚合物可藉化學，生化或物理方法來達到連續訊號放大進而可以肉眼觀測。其長期研究目標是，檢測血液中極低濃度的細菌樣品，並能藉由簡單步驟診斷微生物感染。本研究嘗試證明SHRPSA法的可行性。如同其名，此法利用S-HRP與bBSA 結合使訊號放大。此外，研究包括探討SHRPSA法應用於ELISA的偵測極限，培養時間與試劑濃度比例的優化等。目前我們能證實使用SHRPSA法能夠使IL-7抗原在低濃度6.2 pg/mL下的吸收度放大，且有規律的隨著操作圈數而放大。雖然其背景值稍高，但與整體的放大結果比較後可忽略。現階段我們仍然需要酵素免疫分析儀來檢測吸收度，但在往後的研究我們將嘗試使用poly-HRP解決背景值的問題，降低操作時間，並建立檢測器系統以達到不需儀器即可檢測的目的。 There are many new diseases that are found in the world today, especially in developing countries. An appropriate technology to detect disease for using in resource poor settings remains a challenge. An ideal diagnostic test is affordable, sensitive, specific, simple to perform with minimal training, rapid, robust (stable without refrigeration), and equipment-free. Traditional bacterial detecting technologies, such as plate count technique, Enzyme-linked immunosorbent assay (ELISA), biochemical tests, and the polymerase chain reaction (PCR) are time-consuming and labor-intensive. The main problem with these techniques being performed in developing countries and resource-poor settings is that they rely on instrumentation, expensive reagents, and are labor intensive.The protein conglomeration based amplification method (PCBAM) proposal is a visual detection technique and is apply to identification of samples that is pathogenic microorganisms from human. The core of this developing method is to utilize biotinyled bovine serum albumin (bBSA), and streptavidin-horseradish peroxidase conjugate (S-HRP) to forming polymer-like complexes. The hypothesis is that the complexes would through chemical, biological or physical reaction to achieve continuous signal amplification and can be visually detected in the end. The ultimate goal of PCBAM is to detect low concentration of bacteria in human blood and develop an easy-to-use pathway to determine the microbial infection. In this study, we attempt to demonstrate the feasibility of a SHRPSA method. As its name, this method uses the combination of S-HRP and bBSA to amplify the signal. In addition, we explored the detection limit of ELISA with SHRPSA method, optimization of the incubation time of the conglomeration components. Now we can confirm that the SHRPSA method can enhance the ELISA signal at 6.2 pg/mL of human IL-7 antigen, and have a regular increase in absorbance with the increase of operation cycles. Though the background is an issue that it can be omitted by comparison of overall the amplification results with increasing absorbance. In current stage, we still need to use the enzyme immunoassay analyzer to detect the absorbance. In future studies, we will try to solve the background issue and reduce the total operation time by using poly-HRP and build up a sensor system to achieve the purpose that the analyte can be detected without instruments.

利用蛋白質聚合物達到訊號放大 = Signal Amplification by the Generation of Protein Polymer Networks
林, 伯烜

利用蛋白質聚合物達到訊號放大 = Signal Amplification by the Generation of Protein Polymer Networks / 林伯烜撰 - [高雄市] : 撰者, 2012[民101]. - 115面 ; 圖，表 ; 30公分.
106年4月25日公開含參考書目.
生物檢測streptavidin

利用蛋白質聚合物達到訊號放大 = Signal Amplification by the Generation of Protein Polymer Networks
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