Building-level wastewater surveillance using tampon swabs and RT-LAMP for rapid SARS-CoV-2 RNA detection
Building-level wastewater surveillance using tampon swabs and RT-LAMP for rapid SARS-CoV-2 RNA detection
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DOI：
10.1039/d1ew00496d
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发表时间：
2021-09-23
影响因子：
5
通讯作者：
Bibby, Kyle
中科院分区：
文献类型：
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作者：
Bivins, Aaron;Lott, Megan;Bibby, Kyle
废水中严重急性呼吸综合征冠状病毒2 (SARS-CoV-2) RNA的监测与2019冠状病毒病（COVID-19）病例和社区一级临床检测阳性均有有益的相关性。大学校园废水监测也显示出对COVID-19病例存在和不存在的有希望的预测能力。然而，迄今为止，这种监测最常依赖于复合采样器和反转录定量PCR （RT-qPCR）技术，这限制了废水监测的可及性和可扩展性，特别是在资源匮乏的环境中。在本研究中，我们尝试使用卫生棉条作为被动拭子进行样本收集和逆转录环介导的等温扩增（RT-LAMP），不需要复杂的热循环设备，以检测废水中的SARS-CoV-2 RNA。该工作流程的结果可在样品采集后三小时内获得。RT-LAMP检测的分析灵敏度比rt液滴数字PCR低约20倍。尽管如此，在对所有学生进行独立的每周临床测试的同时，在一项建筑级废水监测活动中，该方法对突发COVID-19病例的三天阳性预测值（PPV）为75%（不包括恢复期病例），当天阴性预测值（NPV）为80%。这些预测值与使用RT-qPCR进行废水监测所报告的结果相当。这些观察结果表明，即使分析灵敏度较低，卫生棉条拭子和RT-LAMP工作流程也提供了一种具有成本效益和快速的方法，即使在资源不足的环境中，也可以用于可扩展的建筑物级废水监测。
Wastewater surveillance for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA has demonstrated useful correlation with both coronavirus disease 2019 (COVID-19) cases and clinical testing positivity at the community level. Wastewater surveillance on college campuses has also demonstrated promising predictive capacity for the presence and absence of COVID-19 cases. However, to date, such monitoring has most frequently relied upon composite samplers and reverse transcription quantitative PCR (RT-qPCR) techniques, which limits the accessibility and scalability of wastewater surveillance, particularly in low-resource settings. In this study, we trialed the use of tampons as passive swabs for sample collection and reverse transcription loop-mediated isothermal amplification (RT-LAMP), which does not require sophisticated thermal cycling equipment, to detect SARS-CoV-2 RNA in wastewater. Results for the workflow were available within three hours of sample collection. The RT-LAMP assay is approximately 20 times less analytically sensitive than RT-droplet digital PCR. Nonetheless, during a building-level wastewater surveillance campaign concurrent with independent weekly clinical testing of all students, the method demonstrated a three-day positive predictive value (PPV) of 75% (excluding convalescent cases) and same-day negative predictive value (NPV) of 80% for incident COVID-19 cases. These predictive values are comparable to that reported by wastewater monitoring using RT-qPCR. These observations suggest that even with lower analytical sensitivity the tampon swab and RT-LAMP workflow offers a cost-effective and rapid approach that could be leveraged for scalable building-level wastewater surveillance for COVID-19 potentially even in low-resource settings.