Abstract:
Wastewater-based epidemiology (WBE) has been an important tool for population surveillance during the COVID-19 pandemic and continues to play a key role in monitoring SARS-CoV-2 infection levels following reductions in national clinical testing schemes. Studies measuring decay profiles of SARS-CoV-2 in wastewater have underscored the value of WBE, however investigations have been hampered by high biosafety requirements for SARS-CoV-2 infection studies. Therefore, surrogate viruses with lower biosafety standards have been used for SARS-CoV-2 decay studies, such as murine hepatitis virus (MHV), but few studies have directly compared decay rates of both viruses. We compared the persistence of SARS-CoV-2 and MHV in wastewater, using 50 % tissue culture infectious dose (TCID50) and reverse transcription quantitative polymerase chain reaction (RT-qPCR) assays to assess infectious virus titre and viral gene markers, respectively. Infectious SARS-CoV-2 and MHV indicate similar endpoints, however observed early decay characteristics differed, with infectious SARS-CoV-2 decaying more rapidly than MHV. We find that MHV is an appropriate infectious virus surrogate for viable SARS-CoV-2, however inconsistencies exist in viral RNA decay parameters, indicating MHV may not be a suitable nucleic acid surrogate across certain temperature regimes. This study highlights the importance of sample preparation and the potential for decay rate overestimation in wastewater surveillance for SARS-CoV-2 and other pathogens.
摘要:
在COVID-19大流行期间,废水流行病学(WBE)一直是人口监测的重要工具,并在国家临床试验计划减少后,继续在监测SARS-CoV-2感染水平方面发挥关键作用。测量废水中SARS-CoV-2衰减曲线的研究强调了WBE的价值,然而,SARS-CoV-2感染研究的高生物安全性要求阻碍了研究.因此,具有较低生物安全标准的替代病毒已用于SARS-CoV-2衰变研究,如鼠肝炎病毒(MHV),但是很少有研究直接比较这两种病毒的腐烂率。我们比较了SARS-CoV-2和MHV在废水中的持久性,使用50%组织培养感染剂量(TCID50)和逆转录定量聚合酶链反应(RT-qPCR)测定来评估感染性病毒滴度和病毒基因标记,分别。传染性SARS-CoV-2和MHV表明相似的终点,然而观察到的早期衰变特征不同,传染性SARS-CoV-2比MHV衰减更快。我们发现MHV是可行的SARS-CoV-2的合适的感染性病毒替代品,但是病毒RNA衰变参数存在不一致,表明MHV可能不是在某些温度范围内的合适的核酸替代品。这项研究强调了样品制备的重要性以及在SARS-CoV-2和其他病原体的废水监测中可能高估腐烂率。