CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, Dutton Park, QLD 4102, Australia.
Department of Civil and Environmental Engineering, Louisiana State University, 3255 Patrick F. Taylor Hall, Baton Rouge, LA 70803, USA.
Sci Total Environ. 2023 Feb 10;859(Pt 1):160072. doi: 10.1016/j.scitotenv.2022.160072. Epub 2022 Nov 8.
In this study, two virus concentration methods, namely Adsorption-Extraction (AE) and Nanotrap® Magnetic Virus Particles (NMVP) along with commercially available extraction kits were used to quantify endogenous pepper mild mottle virus (PMMoV) and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in nucleic acid extracted from 48 wastewater samples collected over six events from eight wastewater treatment plants (WWTPs). The main aim was to determine which workflow (i.e., concentration and extraction methods) produces greater concentrations of endogenous PMMoV and SARS-CoV-2 gene copies (GC) in comparison with each other. Turbidity and total suspended solids (TSS) of wastewater samples within and among the eight WWTPs were highly variable (41-385 NTU and 77-668 mg/L TSS). In 58 % of individual wastewater samples, the log GC concentrations of PMMoV were greater by NMVP workflow compared to AE workflow. Paired measurements of PMMoV GC/10 mL from AE and NMVP across all 48 wastewater samples were weakly correlated (r = 0.455, p = 0.001) and demonstrated a poor linear relationship (r = 0.207). The log GC concentrations of SARS-CoV-2 in 69 % of individual samples were greater by AE workflow compared to NMVP workflow. In contrast to PMMoV, the AE and NMVP derived SARS-CoV-2 GC counts were strongly correlated (r = 0.859, p < 0.001) and demonstrated a strong linear relationship (r = 0.738). In general, the PMMoV GC achieved by the NMVP workflow decreased with increasing turbidity, but the PMMoV GC by the AE workflow did not appear to be as sensitive to either turbidity or TSS levels. These findings suggest that wastewater sample turbidity or suspended solids concentration, and the intended target for analysis should be considered when validating an optimal workflow for wastewater surveillance of viruses.
在这项研究中，我们使用了两种病毒浓缩方法，即吸附-提取（AE）和纳米陷阱®磁性病毒颗粒（NMVP）以及市售的提取试剂盒，来定量从 8 个污水处理厂（WWTP）的 6 次事件中采集的 48 个废水样本中提取的核酸中的内源性辣椒轻斑驳病毒（PMMoV）和严重急性呼吸系统综合征冠状病毒-2（SARS-CoV-2）。主要目的是确定哪种工作流程（即浓缩和提取方法）产生的内源性 PMMoV 和 SARS-CoV-2 基因拷贝（GC）浓度更高。8 个 WWTP 中的废水样本的浊度和总悬浮固体（TSS）高度可变（41-385 NTU 和 77-668 mg/L TSS）。在 58%的个别废水样本中，与 AE 工作流程相比，NMVP 工作流程下的 PMMoV 对数 GC 浓度更高。对所有 48 个废水样本中来自 AE 和 NMVP 的 PMMoV GC/10 mL 的配对测量显示出较弱的相关性（r = 0.455，p = 0.001），并且显示出较差的线性关系（r = 0.207）。在 69%的个别样本中，AE 工作流程下 SARS-CoV-2 的对数 GC 浓度高于 NMVP 工作流程。与 PMMoV 相反，AE 和 NMVP 衍生的 SARS-CoV-2 GC 计数高度相关（r = 0.859，p < 0.001），并且显示出很强的线性关系（r = 0.738）。一般来说，NMVP 工作流程下的 PMMoV GC 随着浊度的增加而降低，但 AE 工作流程下的 PMMoV GC 似乎对浊度或 TSS 水平不敏感。这些发现表明，在验证病毒废水监测的最佳工作流程时，应考虑废水样本的浊度或悬浮固体浓度以及分析的预期目标。
Zotero 插件
