Development and characterization of enzyme colorimetric detection and its applications in biomedical sciences

• Main Authors: Yu-Jung Li, 李岳蓉
• Other Authors: You-Zung Hsieh
• Format: Others
• Language: zh-TW
• Published: 1999
• Online Access: http://ndltd.ncl.edu.tw/handle/17123487750567939846

碩士 === 國立交通大學 === 應用化學系 === 87 === The goals of the research project are on the development of novel methods for enhancing the sensitivity of enzyme colorimetric detection and multiplex detection based on multicolor enzyme colorimetry. Enzyme colorimetric detection is achieved by the enzymatic reaction to create a colour precipitate on the immediate area. The thesis focuses on the dynamic range and limit of detection for both the enzyme colorimetric detection and the signal amplification method by Catalyzed Reporter Deposition(CARD). With the amplification of CARD, studies have been done on the effect of camptothecin(CPT) to the expression profile of human nasopharyngeal cell line(KB cell). The experimental results show that CARD amplification can raise the sensitivity of the technique by sixty folds comparing with enzyme colorimetric detection. Data consistency is achieved at the same time. On the development of multicolor enzyme colorimetric detection, combining different ratios of different haptens in probe labeling can create many probes with different colors. Two different labeling methods, polymerase chain reaction and random-primed labeling, were compared on the labeling efficiency and the correlation of hapten ratios with color . Using random-primed method, 21 probes with distinct colors were successfully created. They can be used to analyze up to 21 samples simultaneously, which greatly increases the efficiency of analysis.