Comparing analytical methods to detect SARS-CoV-2 in wastewater.
Comparing analytical methods to detect SARS-CoV-2 in wastewater.
复制标题
废水中SARS-CoV-2检测方法的比较
DOI：
10.1016/j.scitotenv.2020.143870
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发表时间：
2021-03-01
期刊：
影响因子：
--
通讯作者：
Sánchez G
中科院分区：
文献类型：
--
作者：
Pérez-Cataluña A;Cuevas-Ferrando E;Randazzo W;Falcó I;Allende A;Sánchez G
基于废水的流行病学 (WBE) 已成为评估 2019 年冠状病毒病 (COVID-19) 大流行的可靠策略。最近的出版物表明，废水中的 SARS-CoV-2 检测在技术上是可行的；然而，有许多不同的协议可供使用，并且大多数所应用的方法尚未经过适当的验证。为此，初步评估了浓缩和提取灭活 SARS-CoV-2 及其替代物的不同程序。使用接种了伽马射线照射的 SARS-CoV-2、猪流行性腹泻病毒 (PEDV) 和孟戈病毒 (MgV) 的城市废水来测试铝基吸附沉淀法和聚乙二醇 (PEG) 沉淀方案的浓缩效率。此外，本研究还比较了两种不同的 RNA 提取方法：商业手动旋转柱离心试剂盒和基于磁性二氧化硅珠的自动化方案。总体而言，评估的浓缩方法不会影响伽马射线照射的 SARS-CoV-2 和 MgV 的回收率，而提取方法对 PEDV 则显示出显着差异。伽马射线照射的 SARS-CoV-2、PEDV 和 MgV 的平均回收率分别为 42.9 ± 9.5%、27.5 ± 14.3% 和 9.0 ± 2.2%。当使用自动化系统时，五个基因组 SARS-CoV-2 目标（N1、N2、基因 E、IP2 和 IP4）的检测限 (LoD95%) 范围从 1.56 log 基因组当量 (ge)/mL (N1) 到 2.22 log ge/mL (IP4)；而当使用基于柱的萃取方法时，观察到的值范围在 2.08 (N1) 和 2.34 (E) log ge/mL 之间。对自然污染废水样品中的不同目标进行了评估，N1、N2、E、IP2 和 IP4 的阳性率分别为 91.2%、85.3%、70.6%、79.4% 和 73.5%。我们的基准比较研究表明，铝沉淀法与自动核酸提取相结合代表了一种灵敏度可接受的方法，可以轻松为 SARS-CoV-2 WBE 监测提供感兴趣的结果。
Wastewater based epidemiology (WBE) has emerged as a reliable strategy to assess the coronavirus disease 2019 (COVID-19) pandemic. Recent publications suggest that SARS-CoV-2 detection in wastewater is technically feasible; however, many different protocols are available and most of the methods applied have not been properly validated. To this end, different procedures to concentrate and extract inactivated SARS-CoV-2 and surrogates were initially evaluated. Urban wastewater seeded with gamma-irradiated SARS-CoV-2, porcine epidemic diarrhea virus (PEDV), and mengovirus (MgV) was used to test the concentration efficiency of an aluminum-based adsorption-precipitation method and a polyethylene glycol (PEG) precipitation protocol. Moreover, two different RNA extraction methods were compared in this study: a commercial manual spin column centrifugation kit and an automated protocol based on magnetic silica beads. Overall, the evaluated concentration methods did not impact the recovery of gamma-irradiated SARS-CoV-2 nor MgV, while extraction methods showed significant differences for PEDV. Mean recovery rates of 42.9 ± 9.5%, 27.5 ± 14.3% and 9.0 ± 2.2% were obtained for gamma-irradiated SARS-CoV-2, PEDV and MgV, respectively. Limits of detection (LoD95%) for five genomic SARS-CoV-2 targets (N1, N2, gene E, IP2 and IP4) ranged from 1.56 log genome equivalents (ge)/mL (N1) to 2.22 log ge/mL (IP4) when automated system was used; while values ranging between 2.08 (N1) and 2.34 (E) log ge/mL were observed when using column-based extraction method. Different targets were also evaluated in naturally contaminated wastewater samples with 91.2%, 85.3%, 70.6%, 79.4% and 73.5% positivity, for N1, N2, E, IP2 and IP4, respectively. Our benchmarked comparison study suggests that the aluminum precipitation method coupled with the automated nucleic extraction represents a method of acceptable sensitivity to provide readily results of interest for SARS-CoV-2 WBE surveillance.
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DOI：
10.1016/j.coesh.2020.09.003
发表时间：
2020-09-30
影响因子：
--
作者：
Ahmed W;Bivins A;Bertsch PM;Bibby K;Choi PM;Farkas K;Gyawali P;Hamilton KA;Haramoto E;Kitajima M;Simpson SL;Tandukar S;Thomas K;Mueller JF
通讯作者：
Mueller JF
影响因子：
9.8
作者：
Guerrero-Latorre, Laura;Ballesteros, Isabel;Rios-Touma, Blanca
通讯作者：
Rios-Touma, Blanca
影响因子：
4.4
作者：
Prevost, B.;Lucas, F. S.;Wurtzer, S.
通讯作者：
Wurtzer, S.
DOI：
10.1093/infdis/jiu384
发表时间：
2014-11-01
期刊：
The Journal of infectious diseases
影响因子：
--
作者：
通讯作者：
--
影响因子：
3.7
作者：
Lin CH;Chen YC;Pan TM
通讯作者：
Pan TM