{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T14:54:59Z","timestamp":1785941699079,"version":"3.56.0"},"reference-count":63,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2024,12,2]],"date-time":"2024-12-02T00:00:00Z","timestamp":1733097600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Wastewater-based surveillance (WBS) is a proven tool for monitoring population-level infection events. Wastewater contains high concentrations of inhibitors, which contaminate the total nucleic acids (TNA) extracted from these samples. We found that TNA extracts from raw influent of Berlin wastewater treatment plants contained highly variable amounts of inhibitors that impaired molecular analyses like dPCR and next-generation sequencing (NGS). By using dilutions, we were able to detect inhibitory effects. To enhance WBS sensitivity and stability, we applied a combination of PCR inhibitor removal and TNA dilution (PIR+D). This approach led to a 26-fold increase in measured SARS-CoV-2 concentrations, practically reducing the detection limit. Additionally, we observed a substantial increase in the stability of the time series. We define suitable stability as a mean absolute error (MAE) below 0.1 log10 copies\/L and a geometric mean relative absolute error (GMRAE) below 26%. Using PIR+D, the MAE could be reduced from 0.219 to 0.097 and the GMRAE from 65.5% to 26.0%, and even further in real-world WBS. Furthermore, PIR+D improved SARS-CoV-2 genome alignment and coverage in amplicon-based NGS for low to medium concentrations. In conclusion, we strongly recommend both the monitoring and removal of inhibitors from samples for WBS.<\/jats:p>","DOI":"10.3390\/microorganisms12122475","type":"journal-article","created":{"date-parts":[[2024,12,2]],"date-time":"2024-12-02T09:00:10Z","timestamp":1733130010000},"page":"2475","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Effective Inhibitor Removal from Wastewater Samples Increases Sensitivity of RT-dPCR and Sequencing Analyses and Enhances the Stability of Wastewater-Based Surveillance"],"prefix":"10.3390","volume":"12","author":[{"given":"Nico","family":"Linzner","sequence":"first","affiliation":[{"name":"Laboratory of Berliner Wasserbetriebe, Berliner Wasserbetriebe, 13629 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1280-6138","authenticated-orcid":false,"given":"Alexander","family":"Bartel","sequence":"additional","affiliation":[{"name":"Unit for Surveillance and Epidemiology of Infectious Diseases, State Office for Health and Social Affairs (SOHSA), 10559 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vera","family":"Schumacher","sequence":"additional","affiliation":[{"name":"Laboratory of Berliner Wasserbetriebe, Berliner Wasserbetriebe, 13629 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5328-3652","authenticated-orcid":false,"given":"Jos\u00e9 Horacio","family":"Grau","sequence":"additional","affiliation":[{"name":"Amedes Medizinische Dienstleistungen GmbH, 37081 G\u00f6ttingen, 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Origin","volume":"579","author":"Zhou","year":"2020","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e00103-22","DOI":"10.1128\/cmr.00103-22","article-title":"Wastewater-Based Surveillance as a Tool for Public Health Action: SARS-CoV-2 and Beyond","volume":"37","author":"Parkins","year":"2024","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e13","DOI":"10.1016\/S2666-5247(20)30172-5","article-title":"SARS-CoV-2, SARS-CoV, and MERS-CoV Viral Load Dynamics, Duration of Viral Shedding, and Infectiousness: A Systematic Review and Meta-Analysis","volume":"2","author":"Cevik","year":"2021","journal-title":"Lancet Microbe"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1283","DOI":"10.1056\/NEJMc2016359","article-title":"Saliva or Nasopharyngeal Swab Specimens for Detection of SARS-CoV-2","volume":"383","author":"Wyllie","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"150376","DOI":"10.1016\/j.scitotenv.2021.150376","article-title":"Contribution of SARS-CoV-2 RNA Shedding Routes to RNA Loads in Wastewater","volume":"806","author":"Crank","year":"2022","journal-title":"Sci. Total Environ."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"164495","DOI":"10.1016\/j.scitotenv.2023.164495","article-title":"Monitoring Influenza and Respiratory Syncytial Virus in Wastewater. Beyond COVID-19","volume":"892","author":"Carcereny","year":"2023","journal-title":"Sci. Total Environ."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"149877","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":"ref_8","unstructured":"Henze, M., and Concha, L. (2008). Wastewater Characterization. Biological Wastewater Treatment, IWA Publishing. Chapter 3."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1318","DOI":"10.1128\/aem.59.5.1318-1324.1993","article-title":"Detection of Enteroviruses in Groundwater with the Polymerase Chain Reaction","volume":"59","author":"Abbaszadegan","year":"1993","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1014","DOI":"10.1111\/j.1365-2672.2012.05384.x","article-title":"PCR Inhibitors\u2014Occurrence, Properties and Removal","volume":"113","author":"Schrader","year":"2012","journal-title":"J. Appl. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2009","DOI":"10.1007\/s00216-020-02490-2","article-title":"PCR Inhibition in qPCR, dPCR and MPS-Mechanisms and Solutions","volume":"412","author":"Sidstedt","year":"2020","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Chianese, S., Fenti, A., Iovino, P., Musmarra, D., and Salvestrini, S. (2020). Sorption of Organic Pollutants by Humic Acids: A Review. Molecules, 25.","DOI":"10.3390\/molecules25040918"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"137193","DOI":"10.1016\/j.chemosphere.2022.137193","article-title":"Prospects for Humic Acids Treatment and Recovery in Wastewater: A Review","volume":"312","author":"Zhu","year":"2023","journal-title":"Chemosphere"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"360","DOI":"10.1136\/jcp.44.5.360","article-title":"Inhibitory Effects of Urine on the Polymerase Chain Reaction for Cytomegalovirus DNA","volume":"44","author":"Khan","year":"1991","journal-title":"J. Clin. Pathol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1007\/s12551-020-00620-9","article-title":"Urea-Aromatic Interactions in Biology","volume":"12","author":"Raghunathan","year":"2020","journal-title":"Biophys. Rev."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/S0167-7012(97)00979-2","article-title":"Removal of PCR Inhibitors from Human Faecal Samples through the Use of an Aqueous Two-Phase System for Sample Preparation Prior to PCR","volume":"28","author":"Lantz","year":"1997","journal-title":"J. Microbiol. Methods"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Bartel, A., Grau, J.H., Bitzegeio, J., Werber, D., Linzner, N., Schumacher, V., Garske, S., Liere, K., Hackenbeck, T., and Rupp, S.I. (2024). Timely Monitoring of SARS-CoV-2 RNA Fragments in Wastewater Shows the Emergence of JN.1 (BA.2.86.1.1, Clade 23I) in Berlin, Germany. Viruses, 16.","DOI":"10.3390\/v16010102"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"148834","DOI":"10.1016\/j.scitotenv.2021.148834","article-title":"A Direct Capture Method for Purification and Detection of Viral Nucleic Acid Enables Epidemiological Surveillance of SARS-CoV-2","volume":"795","author":"Mondal","year":"2021","journal-title":"Sci. Total Environ."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"140621","DOI":"10.1016\/j.scitotenv.2020.140621","article-title":"First Detection of SARS-CoV-2 RNA in Wastewater in North America: A Study in Louisiana, USA","volume":"743","author":"Sherchan","year":"2020","journal-title":"Sci. Total Environ."},{"key":"ref_20","unstructured":"Centers for Disease Control and Prevention, National Center for Immunization and Respiratory Diseases (U.S.), and Division of Viral Diseases (2024, November 20). 2019-Novel Coronavirus (2019-nCoV) Real-Time rRT-PCR Panel Primers and Probes, Available online: https:\/\/stacks.cdc.gov\/view\/cdc\/84525."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1111\/j.1467-9868.2007.00646.x","article-title":"Fast Stable Direct Fitting and Smoothness Selection for Generalized Additive Models","volume":"70","author":"Wood","year":"2008","journal-title":"J. R. Stat. Soc. Ser. B (Stat. Methodol.)"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Chen, C., Twycross, J., and Garibaldi, J.M. (2017). A New Accuracy Measure Based on Bounded Relative Error for Time Series Forecasting. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0174202"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"8239","DOI":"10.1021\/acs.est.4c00580","article-title":"Evaluation of the Impact of Concentration and Extraction Methods on the Targeted Sequencing of Human Viruses from Wastewater","volume":"58","author":"Jiang","year":"2024","journal-title":"Environ. Sci. Technol."},{"key":"ref_24","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":"ref_25","doi-asserted-by":"crossref","unstructured":"Belouhova, M., Peykov, S., Stefanova, V., and Topalova, Y. (2023). Comparison of Two Methods for SARS-CoV-2 Detection in Wastewater: A Case Study from Sofia, Bulgaria. Water, 15.","DOI":"10.3390\/w15040658"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1007\/s11270-023-06821-8","article-title":"Assessment of PCR Inhibitor Removal Methods to Monitor Viruses in Environmental Water Samples: DAX-8 Outperforms Competitors","volume":"235","author":"Hamza","year":"2023","journal-title":"Water Air Soil. Pollut."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1863","DOI":"10.4315\/JFP-20-139","article-title":"Assessment of Commercial DNA Cleanup Kits for Elimination of Real-Time PCR Inhibitors in the Detection of Cyclospora Cayetanensis in Cilantro","volume":"83","author":"Assurian","year":"2020","journal-title":"J. Food Prot."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.mimet.2010.12.027","article-title":"Improving qPCR Efficiency in Environmental Samples by Selective Removal of Humic Acids with DAX-8","volume":"85","author":"Schriewer","year":"2011","journal-title":"J. Microbiol. Methods"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3195","DOI":"10.1128\/jcm.30.12.3195-3199.1992","article-title":"The Magnetic Immuno Polymerase Chain Reaction Assay for Direct Detection of Salmonellae in Fecal Samples","volume":"30","author":"Widjojoatmodjo","year":"1992","journal-title":"J. Clin. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1016\/j.mcp.2008.02.003","article-title":"Detection and Quantification of Human and Bovine Noroviruses by a TaqMan RT-PCR Assay with a Control for Inhibition","volume":"22","author":"Scipioni","year":"2008","journal-title":"Mol. Cell Probes"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.soilbio.2016.12.011","article-title":"A Re-Evaluation of Dilution for Eliminating PCR Inhibition in Soil DNA Samples","volume":"106","author":"Wang","year":"2017","journal-title":"Soil Biol. Biochem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"117977","DOI":"10.1016\/j.watres.2021.117977","article-title":"SARS-CoV-2 Wastewater Surveillance in Germany: Long-Term RT-Digital Droplet PCR Monitoring, Suitability of Primer\/Probe Combinations and Biomarker Stability","volume":"210","author":"Ho","year":"2022","journal-title":"Water Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"175687","DOI":"10.1016\/j.scitotenv.2024.175687","article-title":"Estimating Rates of Change to Interpret Quantitative Wastewater Surveillance of Disease Trends","volume":"951","author":"Holcomb","year":"2024","journal-title":"Sci. Total Environ."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Dhiyebi, H.A., Abu Farah, J., Ikert, H., Srikanthan, N., Hayat, S., Bragg, L.M., Qasim, A., Payne, M., Kaleis, L., and Paget, C. (2023). Assessment of Seasonality and Normalization Techniques for Wastewater-Based Surveillance in Ontario, Canada. Front. Public Health, 11.","DOI":"10.3389\/fpubh.2023.1186525"},{"key":"ref_35","first-page":"2200700","article-title":"Patterns of SARS-CoV-2 Circulation Revealed by a Nationwide Sewage Surveillance Programme, the Netherlands, August 2020 to February 2022","volume":"28","author":"Hetebrij","year":"2023","journal-title":"Eurosurveillance"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"113809","DOI":"10.1016\/j.envres.2022.113809","article-title":"Data Modelling Recipes for SARS-CoV-2 Wastewater-Based Epidemiology","volume":"214","author":"Rauch","year":"2022","journal-title":"Environ. Res."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Aberi, P., Arabzadeh, R., Insam, H., Markt, R., Mayr, M., Kreuzinger, N., and Rauch, W. (2021). Quest for Optimal Regression Models in SARS-CoV-2 Wastewater Based Epidemiology. Ijerph, 18.","DOI":"10.3390\/ijerph182010778"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1007\/978-1-60327-353-4_2","article-title":"Overcoming Inhibition in Real-Time Diagnostic PCR","volume":"943","author":"Hedman","year":"2013","journal-title":"Methods Mol. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1111\/j.1556-4029.2009.01245.x","article-title":"A Study of PCR Inhibition Mechanisms Using Real Time PCR","volume":"55","author":"Opel","year":"2010","journal-title":"J. Forensic Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e39","DOI":"10.1093\/nar\/gng039","article-title":"Mechanisms Underlying the Impact of Humic Acids on DNA Quantification by SYBR Green I and Consequences for the Analysis of Soils and Aquatic Sediments","volume":"31","author":"Zipper","year":"2003","journal-title":"Nucleic Acids Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2569","DOI":"10.1007\/s00216-018-0931-z","article-title":"Inhibition Mechanisms of Hemoglobin, Immunoglobulin G, and Whole Blood in Digital and Real-Time PCR","volume":"410","author":"Sidstedt","year":"2018","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"7891","DOI":"10.1128\/AEM.01428-07","article-title":"Evaluation of Removal of Noroviruses during Wastewater Treatment, Using Real-Time Reverse Transcription-PCR: Different Behaviors of Genogroups I and II","volume":"73","author":"Parnaudeau","year":"2007","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"3984","DOI":"10.1128\/aem.58.12.3984-3990.1992","article-title":"Presence of Human Immunodeficiency Virus Nucleic Acids in Wastewater and Their Detection by Polymerase Chain Reaction","volume":"58","author":"Ansari","year":"1992","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1016\/S0166-0934(96)02123-4","article-title":"A Method to Remove Environmental Inhibitors Prior to the Detection of Waterborne Enteric Viruses by Reverse Transcription-Polymerase Chain Reaction","volume":"63","author":"Ijzerman","year":"1997","journal-title":"J. Virol. Methods"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2460","DOI":"10.1021\/acsestwater.2c00306","article-title":"Wastewater-Based Epidemiology for SARS-CoV-2 Biomarkers: Evaluation of Normalization Methods in Small and Large Communities in Southern Germany","volume":"2","author":"Mitranescu","year":"2022","journal-title":"ACS ES T Water"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"105689","DOI":"10.1016\/j.envint.2020.105689","article-title":"Future Perspectives of Wastewater-Based Epidemiology: Monitoring Infectious Disease Spread and Resistance to the Community Level","volume":"139","author":"Sims","year":"2020","journal-title":"Environ. Int."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1007\/s12560-022-09542-z","article-title":"The Inhibition and Variability of Two Different RT-qPCR Assays Used for Quantifying SARS-CoV-2 RNA in Wastewater","volume":"15","author":"Scott","year":"2023","journal-title":"Food Environ. Virol."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Audi, A., AlIbrahim, M., Kaddoura, M., Hijazi, G., Yassine, H.M., and Zaraket, H. (2020). Seasonality of Respiratory Viral Infections: Will COVID-19 Follow Suit?. Front. Public Health, 8.","DOI":"10.3389\/fpubh.2020.567184"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1146\/annurev-virology-012420-022445","article-title":"Seasonality of Respiratory Viral Infections","volume":"7","author":"Moriyama","year":"2020","journal-title":"Annu. Rev. Virol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1728","DOI":"10.1021\/es903216p","article-title":"Antiviral Drugs in Wastewater and Surface Waters: A New Pharmaceutical Class of Environmental Relevance?","volume":"44","author":"Prasse","year":"2010","journal-title":"Environ. Sci. Technol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"103036","DOI":"10.1016\/j.jwpe.2022.103036","article-title":"A Sustainable Approach for the Removal Methods and Analytical Determination Methods of Antiviral Drugs from Water\/Wastewater: A Review","volume":"49","author":"Eryildiz","year":"2022","journal-title":"J. Water Process Eng."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"100076","DOI":"10.1016\/j.nexus.2022.100076","article-title":"Pharmaceutical Wastewater as Emerging Contaminants (EC): Treatment Technologies, Impact on Environment and Human Health","volume":"6","author":"Samal","year":"2022","journal-title":"Energy Nexus"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1186\/s13007-014-0042-6","article-title":"Reverse Transcriptase Droplet Digital PCR Shows High Resilience to PCR Inhibitors from Plant, Soil and Water Samples","volume":"10","author":"Dreo","year":"2014","journal-title":"Plant Methods"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1373\/clinchem.2019.304048","article-title":"Digital PCR-An Emerging Technology with Broad Applications in Microbiology","volume":"66","author":"Salipante","year":"2020","journal-title":"Clin. Chem."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1039\/C6NJ00960C","article-title":"Supramolecular Combinations of Humic Polyanions as Potent Microbicides with Polymodal Anti-HIV-Activities","volume":"41","author":"Zhernov","year":"2017","journal-title":"New J. Chem."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Socol, D.C. (2022). Clinical Review of Humic Acid as an Antiviral: Leadup to Translational Applications in Clinical Humeomics. Front. Pharmacol., 13.","DOI":"10.3389\/fphar.2022.1018904"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1168","DOI":"10.1111\/j.1365-2672.2009.04298.x","article-title":"An Improved Method to Extract RNA from Soil with Efficient Removal of Humic Acids","volume":"107","author":"Wang","year":"2009","journal-title":"J. Appl. Microbiol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1467","DOI":"10.1002\/elps.200700699","article-title":"Interaction of Humic Acids with Human DNA: Proposed Mechanisms and Kinetics","volume":"29","author":"Sutlovic","year":"2008","journal-title":"Electrophoresis"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"16086","DOI":"10.1021\/acs.est.3c05019","article-title":"Binding of Dissolved Organic Matter to RNA and Protection from Nuclease-Mediated Degradation","volume":"57","author":"Chatterjee","year":"2023","journal-title":"Environ. Sci. Technol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1128\/JCM.38.1.345-350.2000","article-title":"Identification and Characterization of Immunoglobulin G in Blood as a Major Inhibitor of Diagnostic PCR","volume":"38","year":"2000","journal-title":"J. Clin. Microbiol."},{"key":"ref_61","unstructured":"European Commission, Joint Research Centre, and Global Water Research Coalition (2024). The International Cookbook for Wastewater Practitioners. Vol. 1, SARS-CoV-2, Publications Office."},{"key":"ref_62","unstructured":"National Academies of Sciences, Engineering, and Medicine (2024). Increasing the Utility of Wastewater-Based Disease Surveillance for Public Health Action: A Phase 2 Report, National Academies Press."},{"key":"ref_63","first-page":"2300595","article-title":"Early Detection of the Emerging SARS-CoV-2 BA.2.86 Lineage Through Integrated Genomic Surveillance of Wastewater and COVID-19 Cases in Sweden, Weeks 31 to 38 2023","volume":"28","author":"Berg","year":"2023","journal-title":"Eurosurveillance"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/12\/12\/2475\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:44:29Z","timestamp":1760114669000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/12\/12\/2475"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12,2]]},"references-count":63,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["microorganisms12122475"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms12122475","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,12,2]]}}}