【作者】 楼凯凯；

【导师】 叶树明；

【作者基本信息】 浙江大学 ， 生物物理学， 2006， 硕士

【摘要】 微生物广泛分布于自然界中,对生态系统的物质交换和能量流动起着不可替代的作用,同时微生物与人类健康有着密切的关系。对微生物的研究关乎整个自然界和人类社会。微生物检测技术为微生物的研究提供了有力的手段。传统的微生物检测方法费时费力,难以满足不同情况下的检测要求,而ATP生物发光法作为一种快速、简便的微生物检测方法,在环境监测、食品卫生、生命科学、医药等领域都有广阔的发展前景。其自动原位检测技术的研究更是国内外的首创,可以解决长期连续监测和极端环境下无法进行人工检测等问题,具有重要意义。 本文从ATP生物发光法的基本原理切入,系统分析了反应中的各种影响因素,包括离子和盐度、pH值、游离态ATP、温度以及荧光素酶活性,得到最佳的pH值范围为7.7～8.0,温度为25℃左右。同时分析了其他几种影响因素对反应的影响程度的大小,并提出相应的措施,去除干扰。掌握并优化ATP生物发光法,将其应用于实际水体微生物量的测量,按照淡水和海水两种情况,分别测量了西湖和海洋的水体微生物量,得到西湖水体微生物的水平分布情况和海洋水体微生物的垂直分布情况。还对赤潮的预警做了可行性研究,认为ATP生物发光法结合自动原位监测仪,可以实现赤潮的预警。最后,作为ATP生物发光法检测微生物量的进一步研究,提出一套可行的自动原位监测仪的设计方案。更多还原

【Abstract】 Microorganisms, which widely exist in nature, play important role in the mass exchange and transfer of energy. There is an affinity between microorganisms and human health. The research of microorganisms concerns the whole nature and human society nearly. The measurement technology for microorganism supports the research of microorganisms as a powerful tool. However, traditional measurement method takes too long time to meet with different conditions. ATP bioluminescence technology, as a rapid and convenient measurement method for microorganisms, has a great prospect in application in the fields of environmental monitoring, food hygiene, life science, medicine, etc. The research of automatic online measurement technology is innovative at home and abroad. It can solve the problems of long-term continual monitoring and measurements in the extreme environments which are not suitable for human activity. Therefore, the research of automatic online measurement technology is extremely significant.This study, based on the elementary reaction principle, analyzed different factors which influenced the ATP bioluminescence reaction, including ions and salinity, pH value, dissociative ATP, temperature and the activity of luciferase. The optimal pH value was between 7.7 and 8.0 and the temperature was about 25 ℃. The affection level of other factors was also analyzed, and solutions were proposed to eliminate the interference. Furthermore, ATP bioluminescence technology was applied in the detection of microorganisms in natural water. Research objects were waters in West Lake and marine in terms of fresh water and salt water. As results, microbialhorizontal distribution of West Lake and microbial vertical distribution of marine were obtained. Moreover, feasibility of forecast to Red Tide was studied. The result indicated that ATP bioluminescence technology, combined with automatic online measure-meter, could realize the forecast to Red Tide. Finally, as further study of the detection of microorganisms by ATP bioluminescence technology, a feasible design of automatic online measure-meter was proposed.更多还原