First environmental surveillance for the presence of SARS-CoV-2 RNA in wastewater and river water in Japan
First environmental surveillance for the presence of SARS-CoV-2 RNA in wastewater and river water in Japan
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DOI：
10.1016/j.scitotenv.2020.140405
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发表时间：
2020-10-01
影响因子：
9.8
通讯作者：
Kitajima, Masaaki
中科院分区：
文献类型：
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作者：
Haramoto, Eiji;Malla, Bikash;Kitajima, Masaaki
基于水的流行病学是了解社区2019冠状病毒病（COVID-19）实际发病率的有力工具，因为COVID-19的病原体严重急性呼吸道综合征冠状病毒2（SARS-CoV-2）可以在感染者的粪便中脱落，无论其症状如何。本研究旨在评估日本山梨县废水和河水中SARS-CoV-2 RNA的存在，使用四种定量和两种巢式PCR检测。在2020年3月17日至5月7日期间，分别从污水处理厂和河流中收集了五次流入和二次处理（氯化前）的废水样品和河水样品。采用两种不同的方法处理200-5000 mL的废水和河水样品：电负性膜涡旋（EMV）法和膜吸附-直接提取RNA法。通过对辣椒轻斑驳病毒RNA的浓度测定，发现EMV法上级膜吸附-直接RNA提取法。采用N_Sarbeco qPCR检测方法，根据EMV方法，在5个二级处理的废水样品中的一个样品中成功检测到SARS-CoV-2 RNA，浓度为2.4 x 10（3）拷贝/L，并对qPCR产物进行了序列确认，而所有进水样品的检测结果均为SARS-CoV-2 RNA阴性。这一结果可能归因于较低过滤体积（200 mL）的进水（4.0 x 10（3）-82 x 10（4）拷贝/L）的检测限高于较高过滤体积（5000 mL）的二级处理废水（1.4 x 10（2）-2.5 x 10（3）拷贝/L）的检测限。与山梨县报告的COVID-19病例相比，没有一个河水样本检测出SARS-CoV-2 RNA阳性。这是日本首次报告在废水中检测到SARS-CoV-2 RNA的研究。(C)2020年，任作家。由爱思唯尔公司出版。这是一篇开放获取的文章，使用CC BY许可证（http：//creativecommons.org/licenses/by/4.0/）。
Wastewater-based epidemiology is a powerful tool to understand the actual incidence of coronavirus disease 2019 (COVID-19) in a community because severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the etiological agent of COVID-19, can be shed in the feces of infected individuals regardless of their symptoms. The present study aimed to assess the presence of SARS-CoV-2 RNA in wastewater and river water in Yamanashi Prefecture, Japan, using four quantitative and two nested PCR assays. Influent and secondary-treated (before chlorination) wastewater samples and river water samples were collected five times from a wastewater treatment plant and three times from a river, respectively, between March 17 and May 7, 2020. The wastewater and river water samples (200-5000 mL) were processed by using two different methods: the electronegative membrane-vortex (EMV) method and the membrane adsorption-direct RNA extraction method. Based on the observed concentrations of indigenous pepper mild mottle virus RNA, the EMV method was found superior to the membrane adsorption-direct RNA extraction method. SARS-CoV-2 RNA was successfully detected in one of five secondary-treated wastewater samples with a concentration of 2.4 x 10(3) copies/L by N_Sarbeco qPCR assay following the EMV method, with sequence confirmation of the qPCR product, whereas all the influent samples were tested negative for SARS-CoV-2 RNA. This result could be attributed to higher limit of detection for influent (4.0 x 10(3)-82 x 10(4) copies L) with a lower filtration volume (200 mL) compared to that for secondary-treated wastewater (1.4 x 10(2)-2.5 x 10(3) copies/L) with a higher filtration volume of 5000 mL. None of the river water samples tested positive for SARS-CoV-2 RNA Comparison with the reported COVID-19 cases in Yamanashi Prefecture showed that SARS-CoV-2 RNA was detected in the secondary-treated wastewater sample when the cases peaked in the community. This is the first study reporting the detection of SARS-CoV-2 RNA in wastewater in Japan. (C) 2020 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).