中文摘要
胰島素是一種多胜肽荷爾蒙，由胰臟製造與分泌並參與醣類代謝，進而提供細胞能量以維持生理功能，當身體出現異常，胰島素分泌不足或是胰島素作用不良時，血糖無法維持平衡，而出現糖尿病。膠體奈米金粒子是在免疫層析檢測試片中最常用來標幟抗體的一種，因為其具有穩定性,且肉眼易觀察之優點。此外，金探針標幟抗體並不需要反應時間、清洗步驟及與酶反應等費時的程序來產生信號。本研究目的為開發檢測人類胰島素之競爭型與三明治型奈米金免疫層析試片，以提供初步檢測胰島素以及胰臟健康情況。實驗結果顯示，此自製式競爭型免疫層析試片能於緩衝溶液與小鼠血清中進行檢測，且不會與其它蛋白質有交叉反應，可檢測範圍為5-100 µg人類胰島素，檢測時間約15-20分鐘。本研究所製備的可拋棄式免疫層析試片具有下列優點：1. 高度專一性與靈敏度；2. 樣品不需前處理；3. 所需樣品量少；4. 操作方便；5. 檢測快速；6. 低成本；7. 沒有交叉反應；8. 不需要任何儀器之使用；9. 試片可長期保存。本研究所開發之試片預期可應用於健康檢查之糖尿病篩檢及胰臟健康狀態之評估。 Abstract
Insulin is a peptide hormone that produced and secreted by the pancreas to involve in regulating glucose metabolism and provide cellular energy to maintain the physiological function. When the body is abnormal, the insulin secretion becomes dysfunctional so that the insulin secretion will be unable to maintain blood sugar balance resulted in diabetes. The colloidal gold nanoparticles are the most commonly used detector reagent in these strips for visualization of signals because of their stability and easy visual observation. Moreover, gold probes do not require time-consuming procedures such as incubations, washing steps and enzymatic reactions to generate a signal.
The main purpose of this study is to develop the competitive and sandwich format which based on nano-gold immunochromatographic strip for detection of insulin. We had successfully developed competitive nano-gold immunochromatographic strip system, the strip can provide diabetes screening and the pancreas health diagnosis.
According to the experimental results, the self-assembled immunochromatographic strip can determine human insulin in PBS and mice serum. This results also showed no cross-reaction with other protein, detect the range of 5-100 μg of human insulin and detection time was about 15-20 minutes. The advantages of self-assembled immunochromatographic strip was highly specific and sensitive, no pretreatment, low sample requirement, easy to operate, low cost, rapid determination, no cross-reaction, long-term preservation, no machine needed etc. In this study, the development of the immunochromatographic strip expected to be used for homecare and point-of-care of diabetes, also assess the health status of the pancreas.
