Kabir Md Pervez, Plaza-Diaz Julio, Mercier Élisabeth, D'Aoust Patrick M, Goodridge Lawrence, Lawal Opeyemi U, Wan Shen, Hegazy Nada, Nguyen Tram, Wong Chandler, Addo Felix, Renouf Elizabeth, Graber Tyson E, Delatolla Robert
Department of Civil Engineering, University of Ottawa, Ottawa, Ontario K1N 1A2, Canada.
Children's Hospital of Eastern Ontario Research Institute, Ottawa, Ontario K1H 8L1, Canada.
ACS ES T Water. 2025 Jul 7;5(8):4472-4481. doi: 10.1021/acsestwater.5c00142. eCollection 2025 Aug 8.
Wastewater genomic surveillance (WWGS) of SARS-CoV-2 is typically performed using influent wastewater, but the approach is challenging due to degradation as well as low target concentrations in wastewater. This could be alleviated by utilizing primary sludge; however, this matrix is prone to sequencing library failures. Our study focuses on developing a robust primary sludge-based SARS-CoV-2 genome sequencing method. The study was conducted using 30 parallel influent wastewater and primary sludge samples collected during three different time periods, under three clinically predominant SARS-CoV-2 Omicron lineages in Ottawa, Canada. Results showed that our approach consistently recovered near-complete (≥90%) SARS-CoV-2 genomes from both influent wastewater and primary sludge samples. Prevalent lineage and single nucleotide variant (SNV) profiles were identical ( > 0.05) between influent wastewater and primary sludge. Further analysis indicated that a similar ( > 0.05) number of rare SNVs were detected between influent wastewater and primary sludge. Overall, our approach enables the sequencing of the most concentrated sources of genetic material within the wastewater matrix, providing valuable insights for public health forecasting of infectious disease prevalence beyond the COVID-19 pandemic.
严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的废水基因组监测(WWGS)通常使用进水废水进行,但由于废水的降解以及目标浓度较低,该方法具有挑战性。利用初沉污泥可以缓解这一问题;然而,这种基质容易导致测序文库失败。我们的研究重点是开发一种基于初沉污泥的稳健的SARS-CoV-2基因组测序方法。该研究使用了在加拿大渥太华三个不同时间段收集的30个平行进水废水和初沉污泥样本,这些样本处于三种临床上占主导地位的SARS-CoV-2奥密克戎谱系之下。结果表明,我们的方法始终能从进水废水和初沉污泥样本中获得近乎完整(≥90%)的SARS-CoV-2基因组。进水废水和初沉污泥之间的流行谱系和单核苷酸变异(SNV)图谱相同(>0.05)。进一步分析表明,进水废水和初沉污泥之间检测到的罕见SNV数量相似(>0.05)。总体而言,我们的方法能够对废水基质中最浓缩的遗传物质来源进行测序,为预测COVID-19大流行之外的传染病流行情况提供有价值的公共卫生见解。
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