The detection and stability of the SARS-CoV-2 RNA biomarkers in wastewater influent in Helsinki, Finland.
The detection and stability of the SARS-CoV-2 RNA biomarkers in wastewater influent in Helsinki, Finland.
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DOI：
10.1016/j.scitotenv.2021.145274
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发表时间：
2021-05-20
期刊：
影响因子：
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通讯作者：
Pitkänen T
中科院分区：
文献类型：
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作者：
Hokajärvi AM;Rytkönen A;Tiwari A;Kauppinen A;Oikarinen S;Lehto KM;Kankaanpää A;Gunnar T;Al-Hello H;Blomqvist S;Miettinen IT;Savolainen-Kopra C;Pitkänen T
以海水为基础的严重急性呼吸道综合征冠状病毒2型（SARS-CoV-2）监测用于监测COVID-19疾病的人口水平流行情况。在许多情况下，由于封锁或分析延迟，废水样品的分析可能只能在长时间储存后进行。在这项研究中，研究了储存条件对废水流入物中SARS-CoV-2病毒RNA拷贝数的影响，并使用逆转录定量PCR（RT-qPCR）检测E-Sarbeco、N2和诺如病毒GII，将其与4 °C、−20 °C和−75 °C下诺如病毒随时间的持续性进行了比较。在芬兰，首次在从芬兰赫尔辛基的Viikinmäki废水处理厂收集的24小时复合进水废水样品中检测SARS-CoV-2 RNA的存在。在2020年4月19日至20日期间采集的废水样本等分试样中检测和定量的SARS-CoV-2 RNA拷贝数在储存29、64和84天后仍然保持惊人的稳定。在储存的样品中，当从预离心的沉淀物（即，流入物的颗粒物质）分析时，SARS β冠状病毒和SARS-CoV-2拷贝数（但不是诺如病毒GII拷贝数）似乎略高于上清液（即，水部分），尽管差异无统计学显著性。此外，当废水中掺入SARS-CoV-2时，在最初的28天内观察到4 °C下的线性衰减，而在-20 °C或-75 °C下的58天内没有可见的衰减。总之，冷冻温度应用于存储时，立即SARS-CoV-2 RNA分析从废水流入是不可能的。除了水部分之外，对样品的颗粒物质的分析可以提高检测频率。
Wastewater-based surveillance of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is used to monitor the population-level prevalence of the COVID-19 disease. In many cases, due to lockdowns or analytical delays, the analysis of wastewater samples might only be possible after prolonged storage. In this study, the effect of storage conditions on the RNA copy numbers of the SARS-CoV-2 virus in wastewater influent was studied and compared to the persistence of norovirus over time at 4 °C, −20 °C, and −75 °C using the reverse-transcription quantitative PCR (RT-qPCR) assays E-Sarbeco, N2, and norovirus GII. For the first time in Finland, the presence of SARS-CoV-2 RNA was tested in 24 h composite influent wastewater samples collected from Viikinmäki wastewater treatment plant, Helsinki, Finland. The detected and quantified SARS-CoV-2 RNA copy numbers of the wastewater sample aliquots taken during 19–20 April 2020 and stored for 29, 64, and 84 days remained surprisingly stable. In the stored samples, the SARS betacoronavirus and SARS-CoV-2 copy numbers, but not the norovirus GII copy numbers, seemed slightly higher when analyzed from the pre-centrifuged pellet—that is, the particulate matter of the influent—as compared with the supernatant (i.e., water fraction) used for ultrafiltration, although the difference was not statistically significant. Furthermore, when wastewater was spiked with SARS-CoV-2, linear decay at 4 °C was observed on the first 28 days, while no decay was visible within 58 days at −20 °C or −75 °C. In conclusion, freezing temperatures should be used for storage when immediate SARS-CoV-2 RNA analysis from the wastewater influent is not possible. Analysis of the particulate matter of the sample, in addition to the water fraction, can improve the detection frequency.