The quantification and trends in concentrations for naturally occurring rotaviruses (RV) and enteroviruses (EV) in untreated sewage in various wastewater systems have not often been compared. There is now greater interest in monitoring the infections in the community including live vaccine efficacy by evaluating untreated sewage. The goals of this study were to 1) survey the concentrations of naturally occurring RV and EV in untreated sewage using a reverse transcription—droplet digital polymerase chain reaction (RT-ddPCR) and 2) investigate the use of a new adsorption elution (bag-mediated filtration system (BMFS) using ViroCap filters) against more traditional polyethylene glycol (PEG) precipitation for virus concentration. Sewage samples were collected from lagoons in Kenya and Michigan (MI), the United States (USA) and from wastewater treatment plants (WWTPs) in the USA. RVs were detected at geometric mean concentrations in various locations, California (CA) 1.31 × 10⁵ genome copies/L (gc/L), Kenya (KE) 2.71 × 10⁴ gc/L and Virginia (VA) 1.48 × 10⁵ gc/L, and EVs geometric means were 3.72 × 10⁶ gc/L (CA), 1.18 × 10⁴ gc/L (Kenya), and 6.18 × 10³ gc/L (VA). The mean RV concentrations using BMFS-ViroCap in split samples compared to PEG precipitation methods demonstrated that the levels were only 9% (#s BMFS/PEG) in the Michigan lagoons which was significantly different (p < 0.01). This suggests that RV concentrations in Kenya are around 1.69 × 10⁶ gc/L. Overall, there was no difference in concentrations for the other sampling locations across the methods of virus recovery (i.e., PEG precipitation and HA filters) using one-way ANOVA (F = 1.7, p = 0.2739) or Tukey–Kramer pairwise comparisons (p > 0.05). This study provides useful data on RV and EV concentrations in untreated sewage in Kenya and the USA. It also highlights on the usefulness of the RT-ddPCR for absolute quantification of RV and EV in sewage samples. The BMFS using ViroCap filters while less efficient compared to the more traditional PEG precipitation method was able to recover RVs and EVs in untreated sewage and may be useful in poor resource settings while underestimating viruses by 1 to 1.5 logs.

各种废水系统中未经处理的污水中天然存在的轮状病毒 (RV) 和肠道病毒 (EV) 的量化和浓度趋势并未经常进行比较。现在人们对通过评估未经处理的污水来监测包括活疫苗效力在内的社区感染产生了更大的兴趣。本研究的目标是 1) 使用逆转录-液滴数字聚合酶链反应 (RT-ddPCR) 调查未经处理的污水中天然存在的 RV 和 EV 的浓度，以及 2) 研究新吸附洗脱（袋-使用 ViroCap 过滤器的介导过滤系统 (BMFS) 对更传统的聚乙二醇 (PEG) 沉淀进行病毒浓缩。从肯尼亚和密歇根 (MI) 的泻湖收集污水样本，美国 (USA) 和来自美国的废水处理厂 (WWTP)。RV 在不同地点的几何平均浓度下检测到，加利福尼亚 (CA) 1.31 × 10⁵基因组拷贝/L (gc/L)，肯尼亚 (KE) 2.71 × 10 ⁴ gc/L 和弗吉尼亚 (VA) 1.48 × 10 ⁵ gc/L，EVs 几何平均值为 3.72 × 10 ⁶ gc/L (CA) 、1.18 × 10 ⁴ gc/L（肯尼亚）和 6.18 × 10 ³ gc/L（VA）。与 PEG 沉淀方法相比，在分割样品中使用 BMFS-ViroCap 的平均 RV 浓度表明，密歇根泻湖中的水平仅为 9% (#s BMFS/PEG)，这是显着不同的 (p < 0.01)。这表明肯尼亚的 RV 浓度约为 1.69 × 10 ⁶克/升。总体而言，使用单向方差分析（F = 1.7，p = 0.2739）或 Tukey-Kramer 成对比较（p > 0.05）。这项研究提供了有关肯尼亚和美国未经处理的污水中 RV 和 EV 浓度的有用数据。它还强调了 RT-ddPCR 在污水样品中 RV 和 EV 的绝对定量方面的有用性。使用 ViroCap 过滤器的 BMFS 虽然与更传统的 PEG 沉淀方法相比效率较低，但能够回收未经处理的污水中的 RV 和 EV，并且可能在资源匮乏的环境中有用，同时将病毒低估了 1 到 1.5 个对数。