Reproducibility and sensitivity of 36 methods to quantify the SARS-CoV-2 genetic signal in raw wastewater: findings from an interlaboratory methods evaluation in the U.S.
Reproducibility and sensitivity of 36 methods to quantify the SARS-CoV-2 genetic signal in raw wastewater: findings from an interlaboratory methods evaluation in the U.S.
复制标题
DOI：
10.1039/d0ew00946f
复制
发表时间：
2021-01-20
期刊：
影响因子：
--
通讯作者：
SARS-CoV-2 Interlaboratory Consortium
中科院分区：
文献类型：
--
作者：
Pecson BM;Darby E;Haas CN;Amha YM;Bartolo M;Danielson R;Dearborn Y;Di Giovanni G;Ferguson C;Fevig S;Gaddis E;Gray D;Lukasik G;Mull B;Olivas L;Olivieri A;Qu Y;SARS-CoV-2 Interlaboratory Consortium
为了应对COVID-19，国际水资源界迅速开发了量化未经处理的废水中SARS-CoV-2基因信号的方法。使用此类方法进行废水监测有可能补充评估社区健康的临床测试。该实验室间评估评估了36个标准操作规程（SOP）的重现性和灵敏度，根据样品浓度方法和是否去除固体分为8个方法组。于2020年8月收集两个未经处理的废水样本，并以基质加标物进行修正（β冠状病毒OC 43），并分发给美国各地的32个实验室。根据项目质量保证计划分析的重复样品在36个SOP中显示出高重现性：80%的回收率校正结果落在±1.15 log 10基因组拷贝/L的条带内，在单个SOP内观察到更高的重现性（标准差为0.13 log 10）。包含固体去除步骤和选择浓缩方法未显示对回收率校正结果的明确、系统性影响。其他方法上的变化（例如，巴氏灭菌、引物组选择和RT-qPCR或RT-dPCR平台的使用）通常导致与其他变异性来源相比的小差异。这些发现表明，多种方法都能够产生可重复的结果，尽管应选择相同的SOP或实验室来跟踪给定设施的SARS-CoV-2趋势。该方法显示了7 log 10范围内的回收效率和检测限，强调回收校正的重要性，并需要考虑方法的灵敏度时，选择废水监测方法。
In response to COVID-19, the international water community rapidly developed methods to quantify the SARS-CoV-2 genetic signal in untreated wastewater. Wastewater surveillance using such methods has the potential to complement clinical testing in assessing community health. This interlaboratory assessment evaluated the reproducibility and sensitivity of 36 standard operating procedures (SOPs), divided into eight method groups based on sample concentration approach and whether solids were removed. Two raw wastewater samples were collected in August 2020, amended with a matrix spike (betacoronavirus OC43), and distributed to 32 laboratories across the U.S. Replicate samples analyzed in accordance with the project’s quality assurance plan showed high reproducibility across the 36 SOPs: 80% of the recovery-corrected results fell within a band of ±1.15 log10 genome copies per L with higher reproducibility observed within a single SOP (standard deviation of 0.13 log10). The inclusion of a solids removal step and the selection of a concentration method did not show a clear, systematic impact on the recovery-corrected results. Other methodological variations (e.g., pasteurization, primer set selection, and use of RT-qPCR or RT-dPCR platforms) generally resulted in small differences compared to other sources of variability. These findings suggest that a variety of methods are capable of producing reproducible results, though the same SOP or laboratory should be selected to track SARS-CoV-2 trends at a given facility. The methods showed a 7 log10 range of recovery efficiency and limit of detection highlighting the importance of recovery correction and the need to consider method sensitivity when selecting methods for wastewater surveillance.
登录
查看更多内容
影响因子：
7.7
作者：
Lago, PM;Gary, HE;Cruz, RG
通讯作者：
Cruz, RG
影响因子：
3.1
作者：
Darnell ME;Subbarao K;Feinstone SM;Taylor DR
通讯作者：
Taylor DR
DOI：
10.1073/pnas.1808798115
发表时间：
2018-11-06
影响因子：
11.1
作者：
Brouwer, Andrew F.;Eisenberg, Joseph N. S.;Eisenberg, Marisa C.
通讯作者：
Eisenberg, Marisa C.
影响因子：
5.4
作者：
Rabenau HF;Cinatl J;Morgenstern B;Bauer G;Preiser W;Doerr HW
通讯作者：
Doerr HW
影响因子：
4.7
作者：
Jureka, Alexander S.;Silvas, Jesus A.;Basler, Christopher F.
通讯作者：
Basler, Christopher F.