Applicability of polyethylene glycol precipitation followed by acid guanidinium thiocyanate-phenol-chloroform extraction for the detection of SARS-CoV-2 RNA from municipal wastewater.
Applicability of polyethylene glycol precipitation followed by acid guanidinium thiocyanate-phenol-chloroform extraction for the detection of SARS-CoV-2 RNA from municipal wastewater.
复制标题
DOI：
10.1016/j.scitotenv.2020.143067
复制
发表时间：
2021-02-20
期刊：
影响因子：
--
通讯作者：
Katayama H
中科院分区：
文献类型：
--
作者：
Torii S;Furumai H;Katayama H
主要浓度和分子过程是实施基于废水的严重急性呼吸综合征冠状病毒2（SARS-CoV-2）流行病学的关键。然而，先前开发的方法针对无包膜病毒进行了优化。很少有研究评估的方法是否适用于有效地回收包膜病毒从各种类型的原污水。本研究的目的是（1）比较一级浓缩[超滤（UF）、电负性膜涡旋（EMV）、和聚乙二醇沉淀（PEG）]和RNA提取方法（使用QIAamp Viral RNA Mini Kit的基于旋转柱的方法和使用TRIzol试剂的酸性硫氰酸胍-苯酚-氯仿提取）（2）检验φ6回收率最高的方法对SARS-CoV-2 RNA检测的适用性。在测试的组合中，PEG+TRIzol提供了最高的φ6回收率，为29.8%至49.8%（几何平均值）。UF + QIAamp Viral RNA Mini Kit提供了第二高的φ6回收率，为6.4%至35.8%。对于UF + TRIzol，观察到相当的φ6回收率（13.8-30.0%）。PEG + QIAamp病毒RNA Mini Kit仅提供1.4%至3.0%的φ6回收率，而大肠杆菌噬菌体MS 2（无包膜病毒的替代物）用PEG + TRIzol重复回收。这表明无包膜替代品（MS 2）不一定验证包膜病毒的有效回收率。EMV + QIAamp Viral RNA Mini Kit在不同类型的原污水中提供了显著不同的φ6回收率（1.6-21%）。然后，改性PEG + TRIzol的适用性进行了检查，在日本东京收集的原污水。在12份抓取样本中，4份SARS-CoV-2 CDC N1和N3检测呈阳性。因此，PEG + TRIzol提供了最高的φ6回收率，并允许从原污水中检测SARS-CoV-2 RNA。比较了初级浓缩和RNA提取的组合。PEG + TRIzol提供最高的φ6回收率，范围为29.8- 49.8%。未包裹的替代品并不一定验证φ6的有效回收。采用改良的PEG+TRIzol方法从日本的原污水中检测到SARS-CoV-2 RNA。
The primary concentration and molecular process are critical to implement wastewater-based epidemiology for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, the previously developed methods were optimized for nonenveloped viruses. Few studies evaluated if the methods are applicable to the efficient recovery of enveloped viruses from various types of raw sewage. This study aims (1) to compare the whole process recovery of Pseudomonas phage φ6, a surrogate for enveloped viruses, among combinations of primary concentration [ultrafiltration (UF), electronegative membrane vortex (EMV), and polyethylene glycol precipitation (PEG)] and RNA extraction methods (spin column-based method using QIAamp Viral RNA Mini Kit and acid guanidinium thiocyanate–phenol–chloroform extraction using TRIzol reagent) for three types of raw sewage and (2) to test the applicability of the method providing the highest φ6 recovery to the detection of SARS-CoV-2 RNA. Among the tested combinations, PEG+TRIzol provided the highest φ6 recovery ratio of 29.8% to 49.8% (geometric mean). UF + QIAamp Viral RNA Mini Kit provided the second highest φ6 recovery of 6.4% to 35.8%. The comparable φ6 recovery was observed for UF + TRIzol (13.8–30.0%). PEG + QIAamp Viral RNA Mini Kit provided only 1.4% to 3.0% of φ6 recovery, while coliphage MS2, a surrogate for nonenveloped viruses, was recovered comparably with PEG + TRIzol. This indicated that the nonenveloped surrogate (MS2) did not necessarily validate the efficient recovery for enveloped viruses. EMV + QIAamp Viral RNA Mini Kit provided significantly different φ6 recovery (1.6–21%) among the types of raw sewage. Then, the applicability of modified PEG + TRIzol was examined for the raw sewage collected in Tokyo, Japan. Of the 12 grab samples, 4 were positive for SARS-CoV-2 CDC N1 and N3 assay. Consequently, PEG + TRIzol provided the highest φ6 recovery and allowed for the detection of SARS-CoV-2 RNA from raw sewage. The combinations of primary concentration and RNA extraction were compared. PEG + TRIzol provided the highest φ6 recovery ranging from 29.8–49.8%. Nonenveloped surrogates did not necessarily validate the efficient recovery of φ6. SARS-CoV-2 RNA was detected by modified PEG+TRIzol from raw sewage in Japan.
登录
查看更多内容
影响因子：
2.5
作者：
Haramoto, E.;Kitajima, M.;Ohgaki, S.
通讯作者：
Ohgaki, S.
影响因子：
9.8
作者：
Haramoto, Eiji;Malla, Bikash;Kitajima, Masaaki
通讯作者：
Kitajima, Masaaki
影响因子：
12.8
作者：
Kazama, Shinobu;Masago, Yoshifumi;Omura, Tatsuo
通讯作者：
Omura, Tatsuo
影响因子：
9.8
作者：
Ahmed, Warish;Bertsch, Paul M.;Kitajima, Masaaki
通讯作者：
Kitajima, Masaaki
影响因子：
5.2
作者：
Gendron, Louis;Verreault, Daniel;Duchaine, Caroline
通讯作者：
Duchaine, Caroline