【作者】 姬晓琴；

【导师】 王晓昌；

【作者基本信息】 西安建筑科技大学 ， 环境科学， 2008， 硕士

【摘要】 目前,由于水环境污染或水系传播引起的病原体感染疾病是世界危害范围最广的问题,引发了许多流行事件,尤其是肠道病毒,引发的案例最多且流行面积最广。因此,对于肠道病毒的研究也就成为当前人们研究的热点问题之一。然而,水体中肠道病毒的含量较少,浓度很低,无法进行直接检测,必须首先从水体中浓缩。另外,传统的细胞培养检测方法,操作繁琐,耗时长,难以在基础实验室进行普及推广。因此,选择一种高效的浓缩方法和快速灵敏的检测方法对环境水体中病毒研究具有很重要的现实意义。本次研究对国内外现有的各种浓缩方法进行了综述比较,并选择了其中的微孔滤膜吸附-洗脱法、滑石粉-硅藻土吸附层吸附-洗脱法、化学絮凝法和滑石粉-硅藻土絮凝沉淀法四种不同的浓缩方法进行比较研究,综合考虑病毒回收率、可靠性等各个方面的因素,选出最佳的浓缩方法为微孔滤膜吸附-洗脱法。本文还对现有的各种肠道病毒检测方法进行了介绍比较,着重阐述了PCR检测方法的原理及其应用研究现状,在RT-PCR方法的基础上建立了水体中肠道病毒的通用引物巢式PCR检测方法,该方法主要是对RT-PCR的产物进行二次扩增,大大提高了检测的特异性和灵敏度。此外,本次研究利用巢式PCR方法对西安市各典型地表水体进行了长期定点连续监测,并对西安市水体中肠道病毒的存在情况和分布特征进行了简单分析。更多还原

【Abstract】 At present, infection of pathogens spreaded by water is the most serious problem in the world, which has caused many epidemics, especially enteroviruses which caused epidemics more than any other viruses, and also the impacted area is the largest. So, study on the enteroviruses become a research hotspot. However, the concentration of the enteroviruses in waters are always very low, and can not be detected directly. Enteroviruses must be concentrated from water to achieve the detection level. Besides, traditional detection method of cell cuture is very complicated, and can not be used universally. Therefore, it is very useful for study on enteroviruses from waters to choose a highly effective concentration method and a sensitive detection method.In this paper, we summarized all the concentration methods used, and we carefully chose four methods to compare,which were microporous filter adsorption-elution method, talcum powder-diatomaceous earth adsorption layer absorption-elution method and talcum powder-diatomaceous earth flocculation precipitation method. Considering about the virus recovery and reliability of the methods, we take the microporous filter adsorption-elution method for the best one. Additionally, we also summarized all the detection methods, and emphasized on the introduction of the principles of PCR and its application. Based on RT-PCR, we build the nested PCR method to detect enteroviruses in waters by universal primer, which amplificated the RT -PCR amplification, and this method could greatly increase the specificity and sensitivity. Furthermore, we deteced the typical surface water in Xi’an city consecutively by nested PCR, and made a analysis of the distrbution characteristic of enteroviruses in Xi’an city.更多还原