Ash K T, Li Y, Alamilla I, Joyner D C, Williams D E, McKay P J, Green B M, Iler C, DeBlander S E, North C M, Kara-Murdoch F, Swift C M, Hazen T C
Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN, United States.
Department of Civil and Environmental Sciences, University of Tennessee, Knoxville, TN, United States.
Front Microbiol. 2023 Feb 9;14:1101205. doi: 10.3389/fmicb.2023.1101205. eCollection 2023.
The COVID-19 pandemic brought about an urgent need to monitor the community prevalence of infection and detect the presence of SARS-CoV-2. Testing individual people is the most reliable method to measure the spread of the virus in any given community, but it is also the most expensive and time-consuming. Wastewater-based epidemiology (WBE) has been used since the 1960s when scientists implemented monitoring to measure the effectiveness of the Polio vaccine. Since then, WBE has been used to monitor populations for various pathogens, drugs, and pollutants. In August 2020, the University of Tennessee-Knoxville implemented a SARS-CoV-2 surveillance program that began with raw wastewater surveillance of the student residence buildings on campus, the results of which were shared with another lab group on campus that oversaw the pooled saliva testing of students. Sample collection began at 8 am, and the final RT-qPCR results were obtained by midnight. The previous day's results were presented to the campus administrators and the Student Health Center at 8 am the following morning. The buildings surveyed included all campus dormitories, fraternities, and sororities, 46 buildings in all representing an on-campus community of over 8,000 students. The WBE surveillance relied upon early morning "grab" samples and 24-h composite sampling. Because we only had three Hach AS950 Portable Peristaltic Sampler units, we reserved 24-h composite sampling for the dormitories with the highest population of students. Samples were pasteurized, and heavy sediment was centrifuged and filtered out, followed by a virus concentration step before RNA extraction. Each sample was tested by RT-qPCR for the presence of SARS-CoV-2, using the CDC primers for N Capsid targets N1 and N3. The subsequent pooled saliva tests from sections of each building allowed lower costs and minimized the total number of individual verification tests that needed to be analyzed by the Student Health Center. Our WBE results matched the trend of the on-campus cases reported by the student health center. The highest concentration of genomic copies detected in one sample was 5.06 × 10 copies/L. Raw wastewater-based epidemiology is an efficient, economical, fast, and non-invasive method to monitor a large community for a single pathogen or multiple pathogen targets.
新冠疫情使得监测社区感染流行情况以及检测严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的存在变得十分迫切。对个体进行检测是衡量病毒在任何特定社区传播情况最可靠的方法，但也是最昂贵且最耗时的。自20世纪60年代科学家开展监测以评估脊髓灰质炎疫苗效果以来，基于污水的流行病学（WBE）就已被使用。从那时起，WBE就被用于监测人群中的各种病原体、药物和污染物。2020年8月，田纳西大学诺克斯维尔分校实施了一项SARS-CoV-2监测项目，该项目从对校园内学生宿舍楼的原污水监测开始，其结果与校园内另一个负责监督学生混合唾液检测的实验室小组共享。样本采集于上午8点开始，最终的逆转录定量聚合酶链反应（RT-qPCR）结果在午夜时分获得。前一天的结果在次日上午8点提交给校园管理人员和学生健康中心。被调查的建筑包括所有校园宿舍、兄弟会和姐妹会，总共46栋建筑，代表了一个超过8000名学生的校内社区。WBE监测依靠清晨的“抓取”样本和24小时混合采样。由于我们只有三台哈希AS950便携式蠕动采样器，我们将24小时混合采样保留给学生人数最多的宿舍。样本经过巴氏消毒，重沉淀物经过离心和过滤去除，然后在RNA提取前进行病毒浓缩步骤。每个样本都通过RT-qPCR检测是否存在SARS-CoV-2，使用美国疾病控制与预防中心（CDC）针对核衣壳靶点N1和N3的引物。随后对每栋楼各区域进行的混合唾液检测降低了成本，并将学生健康中心需要分析的个体验证检测总数降至最低。我们的WBE结果与学生健康中心报告的校内病例趋势相符。在一个样本中检测到的最高基因组拷贝浓度为5.06×10拷贝/升。基于原污水的流行病学是一种高效、经济、快速且非侵入性的方法，可用于监测大型社区中的单一病原体或多个病原体靶点。
