Comparison of viral concentration techniques for native fecal indicators and pathogens from wastewater
Comparison of viral concentration techniques for native fecal indicators and pathogens from wastewater
复制标题
天然粪便指标和废水中病原体的病毒浓缩技术比较
DOI：
10.1016/j.scitotenv.2023.167190
复制
发表时间：
2023
影响因子：
9.8
通讯作者：
Bibby, Kyle
中科院分区：
文献类型：
--
作者：
North, Devin;Bibby, Kyle
病毒病原体通常在环境沃茨中被稀释，在随后的定量或分析之前需要浓缩步骤。历史上，病毒浓缩效率的研究是通过将已知病毒加标至样品中进行的;然而，加标对照品可能与暴露于环境条件的“天然”病毒的行为不同。在本研究中，评估了四种浓缩方法，包括聚乙二醇沉淀（PEG）、脱脂乳絮凝（SMF）、pH下降后通过0.45 μm过滤器（pH）过滤以及使用Amicon过滤器（Amicon）离心，以浓缩废水中的天然病毒靶标。病毒靶点包括指示物（crA病毒和辣椒轻度斑驳病毒）和病原体（腺病毒、诺如病毒GII、人多瘤病毒和SARS-CoV-2）以及细菌标记物（HF 183）。还添加了非天然加标对照，以比较天然和加标回收率。不同靶标和方法的回收率差异很大，范围为0.1 - 39.3%。Amicon方法是回收率的最广泛有效浓度。对于最低滴度的靶标，PEG方法导致最低数量的未检出，SARS-CoV-2的阳性检出率为96.7%，而SMF、pH和Amicon的阳性检出率分别为66.7%、80%和76.7%。选择的非天然加标仅代表少数天然回收率趋势，根据目标和浓度方法而变化，并且始终低于或高于估计的回收率。总体而言，本研究表明，在病毒浓度评价中包括天然靶标并确定特定目标靶标的浓缩方法的效率是有用的。
Viral pathogens are typically dilute in environmental waters, necessitating a concentration step prior to subsequent quantification or analysis. Historically, studies on viral concentration efficiency have been done by spiking known viruses into the sample; however, spike-in controls may not have the same behavior as “native” viruses exposed to environmental conditions. In this study, four concentration methods, including polyethylene glycol precipitation (PEG), skimmed milk flocculation (SMF), pH drop followed by filtration through a 0.45 μm filter (pH), and centrifugation using an Amicon filter (Amicon), were evaluated to concentrate native viral targets in wastewater. Viral targets included both indicators (crAssphage and pepper mild mottle virus) and pathogens (adenovirus, norovirus GII, human polyomavirus, and SARS-CoV-2) in addition to a bacterial marker (HF183). A non-native spike-in control was also added to compare native and spike-in recoveries. Recovery varied widely across targets and methods, ranging from 0.1 to 39.3 %. The Amicon method was the most broadly effective concentration for recovery efficiency. For the lowest-titer target, the PEG method resulted in the lowest number of non-detections, with 96.7 % positive detections for SARS-CoV-2, compared to 66.7 %, 80 %, and 76.7 % positive detections for SMF, pH, and Amicon, respectively. The non-native spike-ins chosen were only representative of a few native recovery trends, varying by both target and concentration method, and consistently under or over-estimated recovery. Overall, this study suggests the utility of including native targets in viral concentration evaluation and determining the efficiency of concentration methods for a specific target of interest.