Development of High Performance DNA Sequencer Using Laser-Induced Capillary Vibration Method

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As can be seen from Human Genome Project, acceleration of analysis time and improvement of sensitivity will promote both fundamental and application study. In recent years, capillary zone electrophoresis (CZE) has attracted much attention as a separation tool for ultramacro amounts of components. however, CZE has no detection method sensitive enough for ultramicro amounts of separated componts, and it has been difficult to apply to ultramicro amounts of materials such as DNA fragments. We proposed capilary vibration induced by laser (CVL) method as a highly sensitive on-column detector for CZE,and attomole amounts of DNA fragments were quantitatively detected using the CVL method. In this study we aimed at developing a novel DNA sequencing method with higher separation efficiency and sensitivity by one order of magnitude by maans of analytical chemistry and molecular biology.This year is the last year of the three year research. In the previous year, we developed pulsed laser-induced stationary wave CVL detection method. We applied the developed method to detection of DNA fragments, and the detection limit of absolute amount was 16 amol. The method was also applied to gene diagnosis, and DNA fragments which was considered to have relation with heart disease or Alzheimer's disease were sensitively detected. These results suggested feasibility of development of gene diagnosis, which is a key to preventive medicine and early detection. Moreover, we applied to high performance immune diagnosis, we investigated on simple method for separating immunologically bound samples and free samples, and developed more sensitive and simpler immunoassay method than the conventional method. Attomole amounts of biological materials were detected with the same or higher sensitivity than the methods using fluorescent tags or radioisotopes. Realization of ultrasensitive detection without labeling process will lead to open a new application field in DNA analysis.