Department of Soil, Water & Environmental Science, Building 38, Room 429, The University of Arizona, Tucson, AZ 85721, USA.
Appl Environ Microbiol. 2011 May;77(10):3500-6. doi: 10.1128/AEM.02705-10. Epub 2011 Mar 25.
The methods used to concentrate enteric viruses from water have remained largely unchanged for nearly 30 years, with the most common technique being the use of 1MDS Virozorb filters followed by organic flocculation for secondary concentration. Recently, a few studies have investigated alternatives; however, many of these methods are impractical for use in the field or share some of the limitations of this traditional method. In the present study, the NanoCeram virus sampler, an electropositive pleated microporous filter composed of microglass filaments coated with nanoalumina fibers, was evaluated. Test viruses were first concentrated by passage of 20 liters of seeded water through the filter (average filter retention efficiency was ≥ 99.8%), and then the viruses were recovered using various salt-based or proteinaceous eluting solutions. A 1.0% sodium polyphosphate solution with 0.05 M glycine was determined to be the most effective. The recovered viruses were then further concentrated using Centricon Plus-70 centrifugal ultrafilters to a final volume of 3.3 (±0.3 [standard deviation]) ml; this volume compares quite favorably to that of previously described methods, such as organic flocculation (~15 to 40 ml). The overall virus recovery efficiencies were 66% for poliovirus 1, 83% for echovirus 1, 77% for coxsackievirus B5, 14% for adenovirus 2, and 56% for MS2 coliphage. In addition, this method appears to be compatible with both cell culture and PCR assays. This new approach for the recovery of viruses from water is therefore a viable alternative to currently used methods when small volumes of final concentrate are an advantage.
用于从水中浓缩肠病毒的方法近 30 年来基本保持不变,最常用的技术是使用 1MDS Virozorb 过滤器,然后进行有机絮凝进行二次浓缩。最近,有一些研究调查了替代方法;然而,许多这些方法在现场使用不切实际,或者具有这种传统方法的一些局限性。在本研究中,评估了 NanoCeram 病毒采样器,这是一种由涂有纳米氧化铝纤维的微玻璃纤维组成的正电褶状微孔过滤器。首先,将 20 升接种水通过过滤器来浓缩测试病毒(平均过滤器保留效率≥99.8%),然后使用各种盐基或蛋白质洗脱液回收病毒。确定 1.0%的多磷酸钠溶液与 0.05 M 甘氨酸的组合是最有效的。然后使用 Centricon Plus-70 离心超滤器将回收的病毒进一步浓缩至最终体积 3.3(±0.3 [标准差])ml;与以前描述的方法(如有机絮凝(~15 至 40 ml))相比,这个体积相当有利。脊髓灰质炎病毒 1 的病毒总回收率为 66%,肠道病毒 1 为 83%,柯萨奇病毒 B5 为 77%,腺病毒 2 为 14%,MS2 噬菌体为 56%。此外,该方法似乎与细胞培养和 PCR 检测都兼容。因此,当需要小体积最终浓缩物时,这种从水中回收病毒的新方法是目前使用的方法的可行替代方法。