Department of Environment and Health, Institute of Health and Environmental Medicine, Key Laboratory of Risk Assessment and Control for Environment & Food Safety , Tianjin 300050, China.
Environ Sci Technol. 2014 Jun 17;48(12):6947-56. doi: 10.1021/es501415m. Epub 2014 Jun 6.
Exposure to various infectious viruses in environmental drinking water can constitute a public health risk. However, it is difficult to detect viruses in water due to their low concentration. In this study, we have developed a novel filter cartridge system containing electropositive granule media (EGM). Viruses present in large volumes of environmental samples were adsorbed onto the EGM, and then recovered by elution and poly(ethylene glycol) (PEG) concentration. To evaluate the system's efficiency in viral recovery, poliovirus (PV-1), a surrogate for enteric viruses, was used to artificially contaminate river water samples which were then assayed by quantitative real-time PCR. To optimize the concentration procedure, the eluent type, water flow rate and properties (e.g., pH, bacterial, and viral loads), were evaluated. The highest virus recovery was obtained by pumping river water at a flow rate of 300 mL/min and then pushing 3 L of an eluent containing 3× broth [1.5% (w/v) NaCl, 3% (w/v) tryptone, 1.5% (w/v) beef powder] with 0.05 mol/L glycine through the filter. Using this procedure, the recovery efficiencies of PV-1 from 10 to 100 L of spiked river water were up to 99%. In addition, this method is virus load and pH dependent. Virus recovery was maximal at a load of between 10(3.5) and 10(5.5) TCID50 and a pH ranging from 5 to 7. The bacterial load in the water has no effect on virus recovery. Different types of viruses and surface water were tested to validate the system's applicability. Results revealed that the EGM filter cartridge was able to concentrate PV-1, human adenoviruses (HAdVs) and noroviruses (HuNoVs) with high efficiency from river, lake, and reservoir water. Furthermore, it showed more efficient recovery than glass wool and 1MDS filters. These data suggest that this system provides rapid and efficient virus recovery from a large volume of natural surface water and, as such, could be a useful tool in revealing the presence of viruses in surface water.
暴露于环境饮用水中的各种传染性病毒可能构成公共卫生风险。然而，由于病毒浓度低，很难在水中检测到病毒。在本研究中，我们开发了一种新型的过滤筒系统，其中包含正电颗粒介质（EGM）。大量环境样品中的病毒被吸附到 EGM 上，然后通过洗脱和聚乙二醇（PEG）浓缩回收。为了评估该系统回收病毒的效率，我们使用脊髓灰质炎病毒（PV-1）作为肠道病毒的替代物，人工污染河水样品，然后通过定量实时 PCR 进行检测。为了优化浓缩程序，我们评估了洗脱液类型、水流速度和特性（例如 pH 值、细菌和病毒负荷）。通过以 300mL/min 的流速泵送河水，然后将含有 3×肉汤[1.5%（w/v）NaCl、3%（w/v）胰蛋白胨、1.5%（w/v）牛肉粉]的 3L 洗脱液以 0.05mol/L 甘氨酸推过过滤器，可获得最高的病毒回收率。使用此程序，从 10 至 100L 已污染河水样品中回收的 PV-1 的回收率高达 99%。此外，该方法依赖于病毒负荷和 pH 值。病毒回收率在 10(3.5)至 10(5.5)TCID50 之间的负荷和 5 至 7 的 pH 值范围内达到最大。水中的细菌负荷对病毒回收率没有影响。我们测试了不同类型的病毒和地表水来验证该系统的适用性。结果表明，EGM 过滤筒能够高效地从河水、湖水和水库水中浓缩 PV-1、人腺病毒（HAdVs）和诺如病毒（HuNoVs）。此外，它的回收率高于玻璃棉和 1MDS 过滤器。这些数据表明，该系统能够从大量天然地表水快速有效地回收病毒，因此可能是揭示地表水病毒存在的有用工具。
