本研究成功製作出具有高靈敏度及高穩定度之快速呈色試劑檢測系統，利用免疫反應的呈色結果作為檢測標的，使免疫呈色反應之偵測往定量方向更加精確。本研究開發之快速呈色檢測系統包含硬體及軟體兩部分，在硬體方面設計出一台呈色判讀儀主體，裝載非指向性發光二極體燈及低成本之網路攝影機，並製作出兩種可替換式載台分別放置檢測卡匣及快速響應矩陣碼。在軟體部分使用Matrix Laboratory (MATLAB)編寫出呈色檢測系統所需的圖形化使用者介面，包含呈色判讀程式本體、快速響應矩陣碼編碼程式、距離量測程式。由實驗結果可以發現本研究所開發之快速呈色檢測系統在對人類絨毛膜促性腺激素檢測試劑量測下，可建立出線性度極佳之濃度與灰階值校正曲線，作為量化分析的依據，該曲線決定係數可達0.98，在人類絨毛膜促性腺激素濃度極限偵測下，可量測最低濃度至6.25 mIU/mL，成功地突破傳統閾值25 mIU/ mL的判定，在重複實驗下仍可保持良好的精準度(標準差<1)。最後將本研究與市售呈色分析儀比較，本研究在反應時間上更快(<10秒)、重複實驗之標準差也均小於市售成色分析儀。

This thesis presents a rapid test system with high sensitivity and stability for human chorionic gonadotropin. The inspection is based on immunochromatographic assay which is widely applied to human chorionic gonadotropin detection. In combination with the immunochromatographic strip, the rapid test system is able to detect and calculate the concentration of the sample. The system is composed of hardware and software parts, and the layout of rapid reader was designed by SolidWorks. The analyzing programs including a main program and two subroutines were compiled by MATLAB. After verification of each parts of the system, we found that the rapid test system successfully detect the minimum 6.25 mIU/mL of human chorionic gonadotropin reagent within several seconds.

[1]A. J. Lapthorn, D. C. Harris, A. Littlejohn, J. W. Lustbader, R. E. Canfield, K. J. Machin, F. J. Morgan and N. W. Isaacs, “Crystal structure of human chorionic gonadotropin,” Nature, 369, pp.455-461, 1994.
[2]J. G. Pierce and T. F. Parsons, “Glycoprotein Hormones: Structure and Function,” Annual Reviews Biochemistry, 50, pp.465-495, 1981.
[3]S. Birken, Y. Maydelman, M. A. Gawinowicz, A. Pound, Y. Liu and A. S. Hartree, “Isolation and characterization of human pituitary chorionic gonadotropin,” Endocrinology, 137, pp.1402-1411, 1996.
[4] http://en.wikipedia.org/wiki/Human_chorionic_gonadotropin.
[5]R. Bellisario, R. B. Carlsen and O. P. Bahl, “Human Chorionic Gonadotropin Linear Amino Acid Sequence of The α Subunit,” Journal of Biological Chemistry, 248, pp.6796-6809, 1973.
[6]R. B. Carlsen, O. P. Bahl and N. Swawinathan, “Human Chorionic Gonadotropin Linear Amino Acid Sequence of The β Subunit,” Journal of Biological Chemistry, 248, pp.6810-6827, 1973.

[7]R. H. Garrett and C. M. Grisham, Biochemistry, Saunders College Publishing, 1995.
[8] 廖國棠，金奈米粒子標記物在免疫分析、DNA 序列分析及微管道晶片系統分析上的應用，國立中山大學化學研究所博士論文，民國九十四年。
[9]E. Engvall and P. Perlmann, “Enzyme-linked immunosorbent assay, Elisa. 3. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes,” Journal of Immunology, 109, pp. 129-135, 1972.
[10] T. Watanabe, Y. Ohkuno, H. Matsuoka, H. Kimura, Y. Sakai, Y. Ohkaru, T. Tanaka, and Y. Kitaura, “Development of a simple whole blood panel test for detection of human heart –type fatty acid-binding protein,” Clinical Biochemistry, 34, pp. 257-263, 2001.
[11]M. Hedenfalk, P. Adlercreutz, and B. Mattiasson, “Modulation of the measuring range of a radioimmunoassay using an organic water two phase system,” Analytica Chimica Acta, 341, pp. 269-274, 1997.
[12]F. Hardy, L. Djavadi-Ohaniance and M. E. Goldberg, “Measurement of antibody/antigen association rate constants in solution by a method based on the enzyme-linked immunosorbent assay,” Journal of immunological methods, 200, pp. 155-159, 1997.
[13]P. Onnerfjord, S. Eremin, J. Emneus and G. Marko-Varga, “Fluorescence polarisation for immunoreagent characterization,” Journal of immunological methods, 213, pp. 31-39, 1998.
[14]J. A. Schmid and A. Billich, “Simple method for high sensitivity chemiluminescence ELISA using conventional laboratory equipment,” BioTechniques, 22, pp. 278, 1997.
[15]C. A. Janeway, P. Travers, M. Walport, and M. J. Shlomchik, Immunobiology, 5th edition, Garland Science, 2001.
[16]http://cht.a-hospital.com/w/hCG
[17]徐敘瑢, 陳憲偉, 光電材料與顯示技術, 五南文化事業出版, 2007.
[18]R. W. McNaught and J. T. France, “Studies of the biochemical basis of steroid sulphatase deficiency: Preliminary evidence suggesting a defect in membrane-enzyme structure,” Journal of Steroid Biochemistry, 13, pp. 363-373, 1980.
[19]G. Harrison, P. Haffey and E. E. Golub, “A nanogram-level colloidal gold single reagent quantitative protein assay,” Analytical Biochemistry, 380, pp.1-4, 2008.
[20]W. N. Zhang and Z. Y. Zhao, “Three-color Data Reproduction Algorithm and Implementation for the Color Image Based on CCD and DSPs,” Computer Engineering, 28, pp.111-113, 2002.
[21] R. Z. Zhou, J. He and Z. L. Hong, “Adaptive Algorithm of Auto White Balance for Digital Camera,” Journal of Computer Aided Design & Computer Graphics, 17, pp.529-533, 2005.
[22]D. Z. Zheng, Z. L. Wu and S. Wang, “Detecting Method of Quantitative Colloidal Gold Test Strip Concentration Based on the DSP Image Processing,” Proc. IEEE 4th International iCBBE Conference (IEEE iCBBE 2010), pp.1-4, 2010.
[23]H. Y. Jiang and M. Du, “Research on the Detection of Gold Immunochromatographic Assay by the Image Histogram Feature Vectors and Fuzzy C-means,” Proc. IEEE 8th International FSKD Conference (IEEE FSKD 2011), pp.467-471, 2011.