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The Efficient and Practical virus Identification System with ENhanced Sensitivity for Solids (EPISENS-S): A rapid and cost-effective SARS-CoV-2 RNA detection method for routine wastewater surveillance
| Title: | The Efficient and Practical virus Identification System with ENhanced Sensitivity for Solids (EPISENS-S): A rapid and cost-effective SARS-CoV-2 RNA detection method for routine wastewater surveillance |
| Authors: | Ando, Hiroki Browse this author | | Iwamoto, Ryo Browse this author | | Kobayashi, Hiroyuki Browse this author →KAKEN DB | | Okabe, Satoshi Browse this author →KAKEN DB | | Kitajima, Masaaki Browse this author |
| Keywords: | Wastewater-based epidemiology | | SARS-CoV-2 | | Pepper mild mottle virus | | COVID-19 | | qPCR | | Quantification method |
| Issue Date: | 15-Oct-2022 |
| Publisher: | Elsevier |
| Journal Title: | Science of the total environment |
| Volume: | 843 |
| Start Page: | 157101 |
| Publisher DOI: | 10.1016/j.scitotenv.2022.157101 |
| Abstract: | Wastewater-based epidemiology has attracted attention as a COVID-19 surveillance tool. Here, we developed a practical method for detecting SARS-CoV-2 RNA in wastewater (the EPISENS-S method), which employs direct RNA extraction from wastewater pellets formed via low-speed centrifugation. The subsequent multiplex one-step RT-preamplification reaction with forward and reverse primers for SARS-CoV-2 and a reverse primer only for pepper mild mottle virus (PMMoV) allowed for qPCR quantification of the targets with different abundances in wastewater from the RT-preamplification product. The detection sensitivity of the method was evaluated using wastewater samples seeded with heat-inactivated SARS-CoV-2 in concentrations of 2.11 x 10(3) to 2.11 x 10(6) copies/L. The results demonstrated that the sensitivity of the EPISENS-S method was two orders of magnitude higher than that of the conventional method (PEG precipitation, followed by regular RT-qPCR; PEG-QVR-qPCR). A total of 37 untreated wastewater samples collected from two wastewater treatment plants in Sapporo, Japan when 1.6 to 18 new daily reported cases per 100,000 people were reported in the city (March 4 to July 8, 2021), were examined using the EPISENS-S method to confirm its applicability to municipal wastewater. SARS-CoV-2 RNA was quantified in 92 % (34/37) of the samples via the EPISENS-S method, whereas none of the samples (0/37) was quantifiable via the PEG-QVR-qPCR method. The PMMoV concentrations measured by the EPISENS-S method ranged from 2.60 x 10(6) to 1.90 x 10(8) copies/L, and the SARS-CoV-2 RNA concentrations normalized by PMMoV ranged from 5.71 x 10(-6 )to 9.51 x 10(-4) . The long-term trend of normalized SARS-CoV-2 RNA concentration in wastewater was consistent with that of confirmed COVID-19 cases in the city. These results demonstrate that the EPISENS-S method is highly sensitive and suitable for routine COVID-19 wastewater surveillance. |
| Type: | article |
| URI: | http://hdl.handle.net/2115/86719 |
| Appears in Collections: | 工学院・工学研究院 (Graduate School of Engineering / Faculty of Engineering) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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