{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T00:18:11Z","timestamp":1790381891840,"version":"4.1.0"},"reference-count":47,"publisher":"Elsevier BV","license":[{"start":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T00:00:00Z","timestamp":1706745600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.elsevier.com\/tdm\/userlicense\/1.0\/"},{"start":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T00:00:00Z","timestamp":1706745600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.elsevier.com\/legal\/tdmrep-license"},{"start":{"date-parts":[[2026,1,5]],"date-time":"2026-01-05T00:00:00Z","timestamp":1767571200000},"content-version":"am","delay-in-days":704,"URL":"http:\/\/www.elsevier.com\/open-access\/userlicense\/1.0\/"},{"start":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T00:00:00Z","timestamp":1706745600000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-017"},{"start":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T00:00:00Z","timestamp":1706745600000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T00:00:00Z","timestamp":1706745600000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-012"},{"start":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T00:00:00Z","timestamp":1706745600000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2024,2,1]],"date-time":"2024-02-01T00:00:00Z","timestamp":1706745600000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-004"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP20H00259"],"award-info":[{"award-number":["JP20H00259"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP20H02284"],"award-info":[{"award-number":["JP20H02284"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP23H01536"],"award-info":[{"award-number":["JP23H01536"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002241","name":"Japan Science and Technology Agency","doi-asserted-by":"publisher","award":["JPMJJR2001"],"award-info":[{"award-number":["JPMJJR2001"]}],"id":[{"id":"10.13039\/501100002241","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002241","name":"Japan Science and Technology Agency","doi-asserted-by":"publisher","award":["JPMJSC20E2"],"award-info":[{"award-number":["JPMJSC20E2"]}],"id":[{"id":"10.13039\/501100002241","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003478","name":"Ministry of Health, Labour and Welfare","doi-asserted-by":"publisher","award":["JPMH20HA2007"],"award-info":[{"award-number":["JPMH20HA2007"]}],"id":[{"id":"10.13039\/501100003478","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003478","name":"Ministry of Health, Labour and Welfare","doi-asserted-by":"publisher","award":["JPMH20HA2009"],"award-info":[{"award-number":["JPMH20HA2009"]}],"id":[{"id":"10.13039\/501100003478","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["elsevier.com","sciencedirect.com"],"crossmark-restriction":true},"short-container-title":["Science of The Total Environment"],"published-print":{"date-parts":[[2024,2]]},"DOI":"10.1016\/j.scitotenv.2023.169746","type":"journal-article","created":{"date-parts":[[2023,12,28]],"date-time":"2023-12-28T04:28:46Z","timestamp":1703737726000},"page":"169746","update-policy":"https:\/\/doi.org\/10.1016\/elsevier_cm_policy","source":"Crossref","is-referenced-by-count":29,"special_numbering":"C","title":["Optimization of the 5-plex digital PCR workflow for simultaneous monitoring of SARS-CoV-2 and other pathogenic viruses in wastewater"],"prefix":"10.1016","volume":"913","author":[{"given":"Bikash","family":"Malla","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sadhana","family":"Shrestha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eiji","family":"Haramoto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"78","reference":[{"key":"10.1016\/j.scitotenv.2023.169746_bb0005","doi-asserted-by":"crossref","DOI":"10.1016\/j.scitotenv.2020.138764","article-title":"First confirmed detection of SARS-CoV-2 in untreated wastewater in Australia: a proof of concept for the wastewater surveillance of COVID-19 in the community","volume":"728","author":"Ahmed","year":"2020","journal-title":"Sci. Total Environ."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0010","doi-asserted-by":"crossref","DOI":"10.1016\/j.scitotenv.2020.139960","article-title":"Comparison of virus concentration methods for the RT-qPCR-based recovery of murine hepatitis virus, a surrogate for SARS-CoV-2 from untreated wastewater","volume":"739","author":"Ahmed","year":"2020","journal-title":"Sci. Total Environ."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0015","doi-asserted-by":"crossref","DOI":"10.1016\/j.watres.2022.118132","article-title":"Evaluation of process limit of detection and quantification variation of SARS-CoV-2 RT-qPCR and RT-dPCR assays for wastewater surveillance","volume":"213","author":"Ahmed","year":"2022","journal-title":"Water Res."},{"issue":"11","key":"10.1016\/j.scitotenv.2023.169746_bb0020","doi-asserted-by":"crossref","first-page":"1871","DOI":"10.1021\/acsestwater.1c00387","article-title":"Comparison of RT-qPCR and RT-dPCR platforms for the trace detection of SARS-CoV-2 RNA in wastewater","volume":"2","author":"Ahmed","year":"2022","journal-title":"ACS ES&T Water"},{"key":"10.1016\/j.scitotenv.2023.169746_bb0025","doi-asserted-by":"crossref","DOI":"10.1016\/j.scitotenv.2021.149877","article-title":"Minimizing errors in RT-PCR detection and quantification of SARS-CoV-2 RNA for wastewater surveillance","volume":"805","author":"Ahmed","year":"2022","journal-title":"Sci. Total Environ."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0030","article-title":"Prevalence of antibiotic resistance genes in drinking and environmental water sources of the Kathmandu Valley","volume":"13","author":"Amarasiri","year":"2022","journal-title":"Nepal. Front. Microbiol."},{"issue":"11","key":"10.1016\/j.scitotenv.2023.169746_bb0035","doi-asserted-by":"crossref","first-page":"1910","DOI":"10.1093\/infdis\/jiaa292","article-title":"The clinical and economic burden of norovirus gastroenteritis in the United States","volume":"222","author":"Bartsch","year":"2020","journal-title":"J. Infect. Dis."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0040","doi-asserted-by":"crossref","first-page":"7754","DOI":"10.1021\/acs.est.0c02388","article-title":"Wastewater-based epidemiology: global collaborative to maximize contributions in the 2 fight against COVID-19","volume":"54","author":"Bivins","year":"2020","journal-title":"Environ. Sci. Technol."},{"issue":"1","key":"10.1016\/j.scitotenv.2023.169746_bb0045","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1038\/s41597-023-02297-7","article-title":"Human viral nucleic acids concentrations in wastewater solids from Central and Coastal California USA","volume":"10","author":"Boehm","year":"2023","journal-title":"Sci. Data"},{"key":"10.1016\/j.scitotenv.2023.169746_bb0050","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1007\/978-3-319-30723-7_3","article-title":"The molecular virology of enteric viruses","author":"Buesa","year":"2016","journal-title":"Viruses Foods"},{"key":"10.1016\/j.scitotenv.2023.169746_bb0055","author":"CDC"},{"key":"10.1016\/j.scitotenv.2023.169746_bb0060","doi-asserted-by":"crossref","DOI":"10.1016\/j.jviromet.2021.114230","article-title":"Assessing sensitivity and reproducibility of RT-ddPCR and RT-qPCR for the quantification of SARS-CoV-2 in wastewater","volume":"297","author":"Ciesielski","year":"2021","journal-title":"J. Virol. Methods"},{"issue":"7","key":"10.1016\/j.scitotenv.2023.169746_bb0065","doi-asserted-by":"crossref","first-page":"3706","DOI":"10.1021\/ac5041617","article-title":"Highly reproducible absolute quantification of Mycobacterium tuberculosis complex by digital PCR","volume":"87","author":"Devonshire","year":"2015","journal-title":"Anal. Chem."},{"issue":"1","key":"10.1016\/j.scitotenv.2023.169746_bb0070","article-title":"Comparison of four digital PCR platforms for accurate quantification of DNA copy number of a certified plasmid DNA reference material","volume":"5","author":"Dong","year":"2015","journal-title":"Sci. Rep."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0080","doi-asserted-by":"crossref","first-page":"893","DOI":"10.1080\/02786826.2010.501351","article-title":"Evaluation of filters for the sampling and quantification of RNA phage aerosols","volume":"44","author":"Gendron","year":"2010","journal-title":"Aerosol Sci. Technol."},{"issue":"23","key":"10.1016\/j.scitotenv.2023.169746_bb2415","doi-asserted-by":"crossref","first-page":"7413","DOI":"10.1128\/AEM.02354-13","article-title":"Occurrence of pepper mild mottle virus in drinking water sources in Japan","volume":"79","author":"Haramoto","year":"2013","journal-title":"Appl. Environ. Microbiol."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0090","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1016\/j.watres.2018.02.004","article-title":"A review on recent progress in the detection methods and prevalence of human enteric viruses in water","volume":"135","author":"Haramoto","year":"2018","journal-title":"Water Res."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0095","doi-asserted-by":"crossref","DOI":"10.1016\/j.scitotenv.2020.140405","article-title":"First environmental surveillance for the presence of SARS-CoV-2 RNA in wastewater and river water in Japan","volume":"737","author":"Haramoto","year":"2020","journal-title":"Sci. Total Environ."},{"issue":"22","key":"10.1016\/j.scitotenv.2023.169746_bb0100","doi-asserted-by":"crossref","first-page":"8604","DOI":"10.1021\/ac202028g","article-title":"High-throughput droplet digital PCR system for absolute quantitation of DNA copy number","volume":"83","author":"Hindson","year":"2011","journal-title":"Anal. Chem."},{"issue":"8","key":"10.1016\/j.scitotenv.2023.169746_bb0105","doi-asserted-by":"crossref","first-page":"1012","DOI":"10.1093\/clinchem\/hvaa125","article-title":"The digital MIQE guidelines update: minimum information for publication of quantitative digital PCR experiments for 2020","volume":"66","author":"Huggett","year":"2020","journal-title":"Clin. Chem."},{"issue":"12","key":"10.1016\/j.scitotenv.2023.169746_bb0110","doi-asserted-by":"crossref","first-page":"1869","DOI":"10.1111\/jgh.12302","article-title":"Novel single-step multiplex real-time polymerase chain reaction assay for simultaneous quantification of hepatitis virus A, B, C, and E in serum","volume":"28","author":"Irshad","year":"2013","journal-title":"J. Gastroenterol. Hepatol."},{"issue":"4","key":"10.1016\/j.scitotenv.2023.169746_bb2410","doi-asserted-by":"crossref","first-page":"1548","DOI":"10.1128\/JCM.41.4.1548-1557.2003","article-title":"Broadly reactive and highly sensitive assay for Norwalk-like viruses based on real-time quantitative reverse transcription-PCR","volume":"41","author":"Kageyama","year":"2003","journal-title":"J. Clin. Microbiol."},{"issue":"3","key":"10.1016\/j.scitotenv.2023.169746_bb2405","doi-asserted-by":"crossref","first-page":"1033","DOI":"10.1128\/AEM.68.3.1033-1039.2002","article-title":"Development of a virus concentration method and its application to detection of enterovirus and Norwalk virus from coastal seawater","volume":"68","author":"Katayama","year":"2002","journal-title":"Appl. Environ. Microbiol."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0120","doi-asserted-by":"crossref","DOI":"10.1016\/j.scitotenv.2021.152033","article-title":"Evaluation of multiple analytical methods for SARS-CoV-2 surveillance in wastewater samples","volume":"808","author":"Kaya","year":"2022","journal-title":"Sci. Total Environ."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0125","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1016\/j.scitotenv.2014.04.087","article-title":"Relative abundance and treatment reduction of viruses during wastewater treatment processes \u2014 identification of potential viral indicators","volume":"488\u2013489","author":"Kitajima","year":"2014","journal-title":"Sci. Total Environ."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0130","doi-asserted-by":"crossref","DOI":"10.1016\/j.scitotenv.2020.139076","article-title":"SARS-CoV-2 in wastewater: state of the knowledge and research needs","volume":"739","author":"Kitajima","year":"2020","journal-title":"Sci. Total Environ."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0135","author":"Kofu City Waterworks and Sewerage Bureau"},{"issue":"3","key":"10.1016\/j.scitotenv.2023.169746_bb0140","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0118833","article-title":"Digital PCR modeling for maximal sensitivity, dynamic range and measurement precision","volume":"10","author":"Majumdar","year":"2015","journal-title":"PLoS One"},{"key":"10.1016\/j.scitotenv.2023.169746_bb0145","doi-asserted-by":"crossref","first-page":"446","DOI":"10.1007\/s12560-019-09398-w","article-title":"Evaluation of human- and animal-specific viral markers and application of CrAssphage, pepper mild mottle virus, and tobacco mosaic virus as potential fecal pollution markers to river water in Japan","volume":"11","author":"Malla","year":"2019","journal-title":"Food Environ. Virol."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0150","doi-asserted-by":"crossref","DOI":"10.1016\/j.scitotenv.2022.158659","article-title":"Application of a high-throughput quantitative PCR system for simultaneous monitoring of SARS-CoV-2 variants and other pathogenic viruses in wastewater","volume":"853","author":"Malla","year":"2022","journal-title":"Sci. Total Environ."},{"issue":"1","key":"10.1016\/j.scitotenv.2023.169746_bb0155","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1016\/j.jviromet.2010.10.018","article-title":"One-step real-time reverse transcription-PCR assays for detecting and subtyping pandemic influenza A\/H1N1 2009, seasonal influenza A\/H1N1, and seasonal influenza A\/H3N2 viruses","volume":"171","author":"Nakauchi","year":"2011","journal-title":"J. Virol. Methods"},{"issue":"3","key":"10.1016\/j.scitotenv.2023.169746_bb0160","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1039\/D0EW00946F","article-title":"Reproducibility and sensitivity of 36 methods to quantify the SARS-CoV-2 genetic signal in raw wastewater: findings from an interlaboratory methods evaluation in the US","volume":"7","author":"Pecson","year":"2021","journal-title":"Environ. Sci. Water Res. Technol."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0165","doi-asserted-by":"crossref","DOI":"10.1016\/j.scitotenv.2020.144215","article-title":"A comparison of SARS-CoV-2 wastewater concentration methods for environmental surveillance","volume":"760","author":"Philo","year":"2021","journal-title":"Sci. Total Environ."},{"issue":"1","key":"10.1016\/j.scitotenv.2023.169746_bb0170","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1373\/clinchem.2019.304048","article-title":"Digital PCR\u2014an emerging technology with broad applications in microbiology","volume":"66","author":"Salipante","year":"2020","journal-title":"Clin. Chem."},{"issue":"3","key":"10.1016\/j.scitotenv.2023.169746_bb0175","doi-asserted-by":"crossref","first-page":"458","DOI":"10.3390\/microorganisms8030458","article-title":"Epidemiological surveillance of norovirus and rotavirus in sewage (2016\u20132017) in Valencia (Spain)","volume":"8","author":"Santiso-Bell\u00f3n","year":"2020","journal-title":"Microorganisms"},{"issue":"1","key":"10.1016\/j.scitotenv.2023.169746_bb2400","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/0166-0934(95)00025-P","article-title":"Methods to remove inhibitors in sewage and other fecal wastes for enterovirus detection by the polymerase chain reaction","volume":"54","author":"Shieh","year":"1995","journal-title":"J. Virol. Methods"},{"key":"10.1016\/j.scitotenv.2023.169746_bb0190","doi-asserted-by":"crossref","DOI":"10.1016\/j.scitotenv.2023.165926","article-title":"Monitoring hand foot and mouth disease using long-term wastewater surveillance in Japan: quantitative PCR assay development and application","author":"Shrestha","year":"2023","journal-title":"Sci. Total Environ."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0195","doi-asserted-by":"crossref","DOI":"10.1016\/j.scitotenv.2023.165270","article-title":"Long-term SARS-CoV-2 surveillance in wastewater and estimation of COVID-19 cases: an application of wastewater-based epidemiology","volume":"896","author":"Shrestha","year":"2023","journal-title":"Sci. Total Environ."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0200","article-title":"Estimation of Norovirus infections in Japan: an application of wastewater-based epidemiology for enteric disease assessment","author":"Shrestha","year":"2023","journal-title":"Sci. Total Environ."},{"issue":"5","key":"10.1016\/j.scitotenv.2023.169746_bb0210","doi-asserted-by":"crossref","first-page":"898","DOI":"10.1111\/j.2041-210X.2012.00215.x","article-title":"Advances in multiplex PCR: balancing primer efficiencies and improving detection success","volume":"3","author":"Sint","year":"2012","journal-title":"Methods Ecol. Evol."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0215","doi-asserted-by":"crossref","DOI":"10.1016\/j.scitotenv.2022.153816","article-title":"Detection of SARS-CoV-2 RNA in wastewater, river water, and hospital wastewater of Nepal","volume":"824","author":"Tandukar","year":"2022","journal-title":"Sci. Total Environ."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0220","doi-asserted-by":"crossref","DOI":"10.1016\/j.envc.2022.100614","article-title":"Pilot study on wastewater surveillance of dengue virus RNA: lessons, challenges, and implications for future research","volume":"9","author":"Thakali","year":"2022","journal-title":"Environ. Challenges"},{"key":"10.1016\/j.scitotenv.2023.169746_bb0225","doi-asserted-by":"crossref","DOI":"10.1016\/j.scitotenv.2022.155663","article-title":"Application of digital PCR for public health-related water quality monitoring","volume":"837","author":"Tiwari","year":"2022","journal-title":"Sci. Total Environ."},{"issue":"2","key":"10.1016\/j.scitotenv.2023.169746_bb0230","article-title":"Comparison of five polyethylene glycol precipitation procedures for the RT-qPCR based recovery of murine hepatitis virus, bacteriophage phi6, and pepper mild mottle virus as a surrogate for SARS-CoV-2 from wastewater","volume":"807","author":"Torii","year":"2022","journal-title":"Sci. Total Environ."},{"issue":"4","key":"10.1016\/j.scitotenv.2023.169746_bb0235","doi-asserted-by":"crossref","first-page":"e497","DOI":"10.1016\/S2214-109X(19)30545-5","article-title":"Global burden of respiratory infections associated with seasonal influenza in children under 5 years in 2018: a systematic review and modelling study","volume":"8","author":"Wang","year":"2020","journal-title":"Lancet Glob. Health"},{"issue":"1","key":"10.1016\/j.scitotenv.2023.169746_bb0240","first-page":"0108","article-title":"RNA viral community in human feces: prevalence of plant pathogenic viruses","volume":"4","author":"Zhang","year":"2006","journal-title":"PLoS Biol."},{"key":"10.1016\/j.scitotenv.2023.169746_bb0245","doi-asserted-by":"crossref","DOI":"10.1016\/j.watres.2022.119560","article-title":"A rapid, high-throughput, and sensitive PEG-precipitation method for SARS-CoV-2 wastewater surveillance","volume":"230","author":"Zheng","year":"2023","journal-title":"Water Res."}],"container-title":["Science of The Total Environment"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.elsevier.com\/content\/article\/PII:S0048969723083766?httpAccept=text\/xml","content-type":"text\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.elsevier.com\/content\/article\/PII:S0048969723083766?httpAccept=text\/plain","content-type":"text\/plain","content-version":"vor","intended-application":"text-mining"}],"deposited":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T16:21:06Z","timestamp":1785946866000},"score":1,"resource":{"primary":{"URL":"https:\/\/linkinghub.elsevier.com\/retrieve\/pii\/S0048969723083766"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2]]},"references-count":47,"alternative-id":["S0048969723083766"],"URL":"https:\/\/doi.org\/10.1016\/j.scitotenv.2023.169746","relation":{},"ISSN":["0048-9697"],"issn-type":[{"value":"0048-9697","type":"print"}],"subject":[],"published":{"date-parts":[[2024,2]]},"assertion":[{"value":"Elsevier","name":"publisher","label":"This article is maintained by"},{"value":"Optimization of the 5-plex digital PCR workflow for simultaneous monitoring of SARS-CoV-2 and other pathogenic viruses in wastewater","name":"articletitle","label":"Article Title"},{"value":"Science of The Total Environment","name":"journaltitle","label":"Journal Title"},{"value":"https:\/\/doi.org\/10.1016\/j.scitotenv.2023.169746","name":"articlelink","label":"CrossRef DOI link to publisher maintained version"},{"value":"article","name":"content_type","label":"Content Type"},{"value":"\u00a9 2023 Elsevier B.V. All rights reserved.","name":"copyright","label":"Copyright"}],"article-number":"169746"}}