Combining ultrafiltration, immunoseparation and analytical microarrays for the monitoring of viruses from large drinking water samples with a volume of 30,000 L

公共和环境卫生保护要求安全饮用水不含致病细菌和病毒，其浓度可能对消费者的健康构成风险。在饮用水系统中，即使是很低的浓度，水媒病毒也会对公共卫生造成危害。然而，由于必要的检测技术还没有得到充分发展，因此在很大程度上缺乏有关病毒浓度与健康风险的定量数据。水中微量低浓度病毒的计数需要一种快速有效的富集方法，以提高后续检测方法的检测限。假设卫生仅根据世卫组织的水质风险评估是安全的，则需要大量的水样本（10，000 L -30，000 L）。到目前为止，还缺少与处理<10 ml的极低样品体积的现代生物分析检测方法相兼容的相应富集技术。因此，根据本提案，应开发一种富集和检测仪器，该仪器将超滤用于大体积的尺寸依赖性富集，免疫分离用于选择性富集至1 mL的最终体积，以及自动化分析微阵列用于快速和同时检测饮用水中的各种病毒。将在田间研究中通过定量噬菌体MS 2和phiX 174验证组合工艺。指示病毒（噬菌体）和感染性病毒（诺如病毒和腺病毒）之间的相关性研究将在新的化学发光流通微阵列芯片阅读器（MCR 3）上进行。

Public and environmental health protection requires safe drinking water free of pathogenic bacteria and viruses in concentrations that can be a risk for consumer´s health. Waterborne viruses can be a public health hazard in drinking water system even in very low concentrations. However, quantitative data relating virus concentrations to health risks are largely lacking because the necessary detection technologies are only insufficiently developed. The enumeration of viruses in water in low concentrations at minute quantities demands a rapid and efficient enrichment methods in order to improve the limit of detection of subsequent detection methods. Assuming that the hygiene is only secure based on the WHO risk assessment for water quality, large sample volumes of water are needed (10,000 L – 30,000 L). So far, corresponding enrichment technologies are missing which are compatible to modern bioanalytical detection methods dealing with very low sample volumes of <10 ml. Therefore, with this proposal an enrichment and detection instrument shall be developed combining ultrafiltration for size dependent enrichment of large volumes, immunoseparation for selective enrichment to an end-volume of 1 mL, and automated analytical microarrays for rapid and simultaneous detection of various viruses in drinking water. The combined process will be validated in field studies by quantification of the bacteriophages MS2 and phiX174. Correlation studies between indicator viruses (bacteriophages) and infectious viruses (noroviruses and adenoviruses) will be performed on the new chemiluminescence flow-through microarray chip reader (MCR 3).