- 學位論文
光纖式粒子電漿共振生化感測平台:利用標定後金奈米棒對植株中胡瓜嵌紋病毒之定量
Fiber-Optic Particle Plasmon Resonance Sensor: Detection of Cucumber mosaic virus by gold nanorods-modified immunosensor
摘要
本研究開發以金奈米棒應用於光纖粒子電漿共振(Fiber-Optic Particle Plasmon Resonance，FO-PPR)生化感測器做為檢測平台，並以胡瓜嵌紋病毒(Cucumber mosaic virus)作為檢測樣品，以評估本系統之應用性。目前FO-PPR以球狀奈米金做為奈米感測器為主，在許多文獻中也證實具有良好的靈敏性與再現性。但是使用球狀金奈米面對一些具有可見光吸收之檢體，造成光譜干擾，進而使得實驗產生誤差。研究將以金奈米棒做為取代，利用金奈米棒可調式紅外光區，將可有效避免其光譜干擾。 以1%MPTMS(mercaptopropyl)-trimethoxy-silane)修飾上光纖後，浸泡在二次離心金奈米棒溶液120分鐘，可達到19.82%覆蓋率。利用混合自組裝單層修飾金奈米棒並與CMV-抗體進行做鍵結，對於純化CMV進行感測，利用CMV與CMV-抗體特異性結合，將造成光纖中光訊號強度改變， 建立以FOPPR與金奈米棒檢測CMV之平台。本檢測對於CMV之偵測極限可達19.7 ng/mL，對實際植物檢體檢測結果與酵素連結免疫吸附法(Enzyme-linked immunosorbent assay，ELISA)所檢出之結果符合，印證FO-PPR應用於植物病毒CMV之檢測具有實測之效果，並且檢定方法相較ELISA快速且免標定。
並列摘要
Cucumber mosaic virus (CMV) is a plant pathogenic virus. This virus has a worldwide distribution and a very wide host range. In fact it has the reputation of having the widest host range of any known plant virus. There are several methods for the determination of CMV, mainly using enzyme-linked immunosorbent assay (ELISA) and reverse transcription- Polymerase Chain Reaction (RT-PCR) techniques. In this study, one simple and label-free biosensing method has been developed for CMV. The biosensing technique, fiber-optic particle plasmon resonance (FOPPR) sensor, is based on gold nanoparticles-modified optical fiber which has been demonstrated to exhibit high sensitivity. However, using gold nanospheres as the sensing material in FOPPR sensor is interfered by the colored matrix in some samples, leading to experimental error and poor reproducibility. Using Gold nanorods as the sensing material in FOPPR sensor avoids the absorption interference in the visible region by the colored matrix in a sample. To implement the FOPPR sensor based on gold nanorods for biosensing of CMV, the gold nanorod surface is modified by a mixed self-assembled monolayer for further conjugation of anti-CMV antibody and minimization of nonspecific adsorption. Upon binding of CMV to immobilized anti-CMV on the gold nanorod surface, the absorbance of the gold nanorod layer on the optical fiber changes and the signal change is enhanced through multiple total internal reflections along the optical fiber. Results show that the detection of CMV in plant fluid by this sensor agrees with the clinically accepted ELISA method but a much shorter analysis time is required.