臺灣博碩士論文加值系統

設計出高穩定性、靈敏性、快速性、便利性的生物檢測平台是所
有從事相關生物檢測研究人員共同的理念。本篇論文在於是利用石英
晶體微天平做為一腸炎弧菌與乳癌相關檢測平台並藉由了解在此生
物感測器中，將其各項步驟的最適化條件、動力學情形、表面分子排
列結構等，確定出可讓核酸、蛋白質等各種具生物辨識力之生物探針
穩定固定的生物檢測平台。
整體來說，金表面鍵結出官能基方式以電漿接枝法的方式效能最
佳。而使用PEI與GA化學鍵結的方式，可使晶片穩定性提高10 倍。
隨著GA 接枝量提升，核酸探針的鍵結量呈線性上升。然而從百分比
的角度來看，發現隨著GA接枝量提升，核酸探針鍵結在GA的披覆量
反而略為下降。
核酸探針固定在晶片的比例約2%，雜交反應固定量最高可達到
探針固定量的18%，而反應時間僅需十分鐘。晶片的最低靈敏度為
10μg，而達到飽和濃度約為50μg，頻率變化量約45Hz。而另一部
份本文嘗試將抗體固定上晶片雜交反應，初步的結果顯示亦是可行的
方式，且抗體具有活性可進行抗原的專一性辨識。以上結果顯示本文
所設計石英晶體生物檢測平台以達到初步可供檢檢測的效果。

All of the bio-researcher would like to design the detection
bioplatform that be more stable､sensitive､faster､convenient. The paper
is based on using quartz crystal microbalance to research Vibrio
parahamolyticus and breast cancer.
The whole to say, the method of Plasma Graft is the beat efficient to
immobilize functionality on the golden surface. Then it will be more
stability 10 times by using the method of PEI and GA chemistry immobili
-zation way. As the GA’s move up, the DNA probe immobilization
quantity lies to linear ascension.
The ratio of DNA probe immobilization quantity is 2 percent on the
chip, then the best hybridization immobilization quantity is up to 18
percent of the DNA probe immobilization quantity, and the reaction time
only 10 minute. Another part of paper results is want to immobili -zation
antibody and hybridization on the chip, the accidence test is feasible, and
antibody still had activity to recognize specific antigen. The
above-mentioned results to show the paper design’s bio-QCM platform
can be accomplished accidence detectable result.

ENGLISH ABSTRACT……………………………………………..……i
CHINESE ABSTRACT…….....……………...……………...…………..ii
ACKNOWLEDGEMENTS…………...………...………………………iii
TABLE OF CONTENTS…..………………………………...………….iv
LIST OF FIGURES…....………………………………..…………..…...x
LIST OF TABLES……………….……………………...…..………….xiv
NOMENCLATURE……...…………………………..……….….....….xvi
NOMENCLATURE………………………………….….…..……xvi
UNIT SYMBOL…………………………………………...…….xvii
SUPERSCRIPT………………………...…………………...….. xviii
CHAPTER
I INTRODUCTION…...……………....………........…………..………1
1.1 Experiment motive…………………………...………………1
1.2 Experiment purpose.……………………………………...…..4
1.3 Experiment structure....……........…………………..…….….6
1.4 Brief introduction of every chapter…………………………..9
v
II DOCUMENT RETROSPECT….….……………….......….……….11
2.1 Brief introduction of detecting device of living beings……...11
2.1.1 Define and make up……………………………..…..…12
2.1.2 Genesis and develop……………………...……………17
2.1.3 Kind and classification…………………………….…...22
2.1.4 Design principle……………………………..…………26
2.1.5 Application…………………………………..…………30
2.1.6 The assessment of the market and present situation…...32
2.1.7 In the clinical advantage that examines…………..……36
2.2 Quartz crystal little day brief introduction of balance…...…..38
2.2.1 Piezoelectricity phenomenon………………….………39
2.2.2 Piezoelectricity of the quartz crystal…………….….…42
2.2.3 The balance detects and examines the theory way on
quartz piezoelectricity little day……………….………49
2.2.4 The balance detects the way of examining to revise on
quartzy piezoelectricity little day under the liquid
state……………………………………………..…..…52
2.2.5 The balance detects the experiment step and way
examined to revise on quartz piezoelectricity little day
vi
under the liquid state……………………..……..….….56
2.2.6 The surface of the regular biological component is
decorated………………………………..………..……61
2.2.7 Research that the balance is applied to detect of little day
of quartz piezoelectricity……………….………..……63
2.3 Brief introduction of the enteritis Vibrio…………………….65
2.3.1 Cause pathogens poisonned by food……………..……66
2.3.2 Distributed and seasonality……………………………67
2.3.3 Type attitude and characteristic…………………….…69
2.3.4 Difference type of the fungus body……………...……71
2.3.5 Pathogenic strength……………………………………73
2.3.6 Pathology characteristic………………………….……75
2.3.7 Appraise the way………………………………...…….76
2.3.8 The food hygiene is safe………………………………79
2.4 Brief introduction of the breast cancer………………....……81
2.4.1 Epidemiology of the breast cancer……………….……82
2.4.2 Treat the new development of the breast cancer………84
2.4.3 Firefly's mere normal position hybridization law……..86
III MATERIALS AND METHODS......................................…..…….88
vii
3.1 Materials………………………….…………..……….……88
3.1.1 Chemicals and reagents………………………….........88
3.1.2 Bacteria strains……………………….…………...…..89
3.1.3 Primer secquences…………………………..…..…….89
3.1.4 Chip size………………………………………....……89
3.2 Experiment machine…………………...……….…………..90
3.2.1 Generally biochemical machine…………………...….90
3.2.2 Liquid quartz crystal sensing system……………..…..91
3.3 Methods and process………………….……….………..….92
3.3.1 Bacteria Experiment……..………………………....…92
3.3.2 Polyacrylamide Gel Preparation……………………....95
3.3.3 Pretreatment of QCM electrode surface………………97
3.3.4 Liquid QCM experiment…………………....…..…...100
3.3.5 Chip reused experiment………………………...……105
3.4 Experiment design explain…………………….……….…106
IV EXPERIMENT RESULTS……………………………...…...….112
4.1 The best pretreatment of QCM………………..……….….112
4.1.1 Blank chip stability experiment….…………………..113
4.1.2 The difference chip surface methods to the
viii
immobilization quantity on a crystal suface
experiment…………………………………………...117
4.1.3 The influence of difference chemical reaction condition
to the immobilization quantity on a crystal suface
experiment…………………..…………………..…...119
4.2 Building biochip with Vibrio parahaemolyticu DNA
probe……………………………………………….…..….123
4.2.1 The influence of difference DNA probe concentration to
the immobilization quantity on a chip suface..........…123
4.2.2 The influence of different buffer to the DNA probe
hybridization on the chip surface………..……....…..130
4.2.3 The influence of different procedure condition to DNA
probe hybridization immobilization quantity………..132
4.2.4 Mimic experiment by DNA probe chip…………..….140
4.2.5 DNA probe chip reused experiment……………..…..146
4.3 Building antibody breast cancer test biochip……………...148
4.3.1 Accomplish antibody probe biochip…………...…….148
4.3.2 The situation about different antibody probe
immobilization experiment…………………………..150
ix
V DISCUSSION AND CONCLUSIONS….....................................152
5.1 Results and discussion……………………………...……152
5.1.1 The discussion of chip stabilit……..………..…...…..152
5.1.2 The discussion of best treatment steps at DNA chip...159
5.1.3 The discussion of DNA probe hybridizationin the
different influence……………………………..…….169
5.1.4 The discussion of antibody probe hybridizationin
influence………………………….…………..……...173
5.1.5 Estimating the molecular structure on the QCM
surface……………………………………………….174
5.2 Experiment conclusion……………………………...……..175
VI RECOMMENDATION FOR FUTURE........................................177
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