腸病毒七十一型(Enterovirus 71)對世界各地的幼童而言是一種重要的致病原且比其他非小兒麻痺腸病毒具有高致病率及致死率，其感染屬於神經性症狀,受感染的病童平均會在三天內惡化成重症。過去的臨床確認檢驗方式需要先作病毒培養再進行病毒分離和藉由反轉錄聚合酶鏈鎖反應(RT-PCR)，這些檢驗流程趕不上惡化成重症的時間，無法達到即時診斷出EV 71並作後續的醫療處置和疾病控管。在過去的文獻中，多晶矽奈米線場效電晶體可被製成且具有高靈敏度、無需標誌且即時偵測生物分子的生化感測器。將特定腸病毒的DNA序列有專一性的單股DNA序列先固定在多晶矽奈米線場效電晶體表面，將互補和非互補的DNA序列流過奈米線場效電晶體表面，能對DNA序列作專一性辨識而兩互補的DNA序列產生雜交反應使奈米線場效電晶體的導電度產生變化，最低濃度可偵測到fM (femto-molar,10-15M)。此結果表示多晶矽奈米線場線電晶體具有高靈敏度、無需標誌且可即時偵測的潛能，此特性可發展成生物感測系統用來偵測腸病毒的感染型別並應用於其他傳染病篩檢。

Enterovirus 71 (EV 71) is an important pathogen that causes higher morbidity and mortality in children around the world than those of other non-Polio enteroviruses. Its infection is neurotropic and even followed by rapid deterioration within average 3 days. The standard clinical methods for EV 71 identification require virus isolation in cell culture and reverse transcriptase polymerase chain reactions (RT-PCR). Virus isolation requires 5-10 days and hinders the subsequent treatment and disease control.
Poly silicon nanowire field effect transistor has been shown to function as transducer for high sensitive, label-free and real-time biosensing to detect enterovirus infection. The selectivity of target for detection can be achieved by surface modification on NWFET. In our research, specific antibody or DNA sequences that recognize capsid protein or nucleic acid will be immobilized on poly Si NWFET. Currently, we are able to distinguish between EV 71 and CA 16 DNAs by real-time electrical analysis. It will be a promising biosensor for rapid diagnosis of EV 71 or other epidemic diseases.

Abstract (Chinese) i
Abstract (English) iii
Acknowledgement iv
Contents v
Contents of Tables vii
Contents of Figures viii
Abbreviations ix
I Introduction 1
1-1 Enterovirus 1
1-1.1 Introduction of Enterovirus 1
1-1.2 Transmission pathway 2
1-1.3 Epidemiology 3
1-1.4 Current clinical diagnosis methods 3
1-1.4.1 Enterovirus isolation in cell culture 4
1-1.4.2 Enterovirus antisera neutralization test (NT) 4
1-1.4.3 Immunofluorescence assay (IFA) 5
1-1.4.4 Reverse transcription polymerase chain reaction (RT-PCR) 5
1-2 Applying poly crystalline silicon nanowire field effect transistor 6
II Materials and methods 8
2-1 Experimental materials 8
2-2 Instruments 11
2-3 Poly crystalline silicon NWFET fabrication process 11
2-4 Microfluidic system 12
2-5 Surface modification 13
2-6 Sample transport at kinetic equilibrium 14
2-7 Liquid phase electrical measurement 14
III Results and discussion 16
3-1 Nanowire chip selection in dry air condition 16
3-2 Biosensing of non-immobilized semiconductor device 16
3-3 Device characteristic verification in liquid phase 17
3-4 Enterovirus 71 (EV 71) DNA Biosensing 18
3-5 EV 71 DNA biosensing after hot water washed 20
3-6 CA 16 DNA Biosensing 20
IV Summary and Perspective 22
V Reference 23
VI Tables and Figures 30

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