Sathyanarayana Shivaprasad H, Robins Ashlee A, Toledo Diana M, Gallagher Torrey L, Tsongalis Gregory J, Hubbard Jacqueline A, Lefferts Joel A, Martin Isabella W
Department of Pathology and Laboratory Medicine, Dartmouth Health, Lebanon, New Hampshire, USA.
The Broad Institute at MIT and Harvard, Cambridge, Massachusetts, USA.
Microbiol Spectr. 2025 Apr;13(4):e0249024. doi: 10.1128/spectrum.02490-24. Epub 2025 Feb 25.
Wastewater-based surveillance (WBS) can track the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in communities. Laboratory methods for this testing involve labor-intensive, multi-step processes. This study assessed the feasibility of performing WBS with an off-label use of an automated commercial SARS-CoV-2 assay that had received Emergency Use Authorization for human diagnostic testing from the United States Food and Drug Administration (FDA EUA). Twenty-four-hour composite samples of primary influent wastewater from seven municipalities in New Hampshire and Vermont were collected between September 2020 and February 2021, and were centrifuged upon receipt. An aliquot of fresh supernatant was immediately tested with the Abbott 2000 RealTie SARS-CoV-2 assay (Abbott Molecular, Des Plaines, IL, USA). Corresponding aliquots were then stored at -80°C until they were thawed, polyethylene glycol (PEG) concentrated, and tested by two PCR-based laboratory-developed tests (LDTs). Wastewater samples (103) were tested with successful detection of SARS-CoV-2 viral RNA by all three methods. Bland-Altman analysis showed overall concordant results with a bias of -0.13 and -0.42 log copies/mL detected by the FDA EUA assay compared to the LDTs. Specimen stability assessment demonstrated a decrease of 33.9% measurable viral RNA after three freeze-thaw cycles. SARS-CoV-2 detection in wastewater using an FDA EUA assay on an automated commercial testing platform performed comparably but with more efficient workflow when compared to two LDTs. This sample-to-answer automated method could save time and labor for surveillance testing, but further validation of its ability to quantitate SARS-CoV-2 viral RNA is necessary.IMPORTANCEThis proof-of-principle study evaluates an off-label use of an automated United States Food and Drug Administration (FDA) Emergency Use Authorization (EUA) severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) human diagnostic assay for wastewater surveillance. Compared to standard, labor-intensive, multi-step methods currently in use for wastewater surveillance testing, an off-label use of an FDA EUA assay on an automated platform offers a sample-to-answer testing requiring less labor and a faster turnaround time.
基于废水的监测（WBS）能够追踪严重急性呼吸综合征冠状病毒2（SARS-CoV-2）在社区中的传播情况。用于此项检测的实验室方法涉及劳动强度大的多步骤流程。本研究评估了将一种已获得美国食品药品监督管理局（FDA）紧急使用授权用于人类诊断检测的自动化商用SARS-CoV-2检测方法超说明书使用以进行WBS的可行性。在2020年9月至2021年2月期间，收集了新罕布什尔州和佛蒙特州七个城市的24小时初级进水废水混合样本，并在收到样本后进行离心处理。取一份新鲜上清液立即使用雅培2000 RealTie SARS-CoV-2检测方法（雅培分子公司，美国伊利诺伊州德斯普兰斯）进行检测。然后将相应的样本分装在-80°C下保存，直至解冻、用聚乙二醇（PEG）浓缩，并通过两种基于聚合酶链反应（PCR）的实验室自行开发的检测方法（LDTs）进行检测。对103份废水样本进行了检测，所有三种方法均成功检测到了SARS-CoV-2病毒RNA。布兰德-奥特曼分析显示，与LDTs相比，FDA紧急使用授权检测方法检测到的偏差为-0.13和-0.42 log拷贝/毫升，总体结果一致。样本稳定性评估表明，经过三个冻融循环后，可测量的病毒RNA减少了33.9%。与两种LDTs相比，在自动化商用检测平台上使用FDA紧急使用授权检测方法检测废水中的SARS-CoV-2，结果相当，但工作流程更高效。这种从样本到结果的自动化方法可为监测检测节省时间和人力，但有必要对其定量SARS-CoV-2病毒RNA的能力进行进一步验证。重要性本项原理验证研究评估了一种美国食品药品监督管理局（FDA）紧急使用授权（EUA）的自动化严重急性呼吸综合征冠状病毒2（SARS-CoV-2）人类诊断检测方法在废水监测中的超说明书使用情况。与目前用于废水监测检测的标准、劳动强度大的多步骤方法相比，在自动化平台上超说明书使用FDA紧急使用授权检测方法可提供一种从样本到结果的检测，所需人力更少，周转时间更快。
