ArticleWater research2022
Evaluation of process limit of detection and quantification variation of SARS-CoV-2 RT-qPCR and RT-dPCR assays for wastewater surveillance.
Article in Water research, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed, 84 citations in OpenAlex.
- Monkeypox Virus Surveillance in Wastewater, North Carolina, USA, October 2023-May 2025.Emerging infectious diseases · 2026Article
- Bio-Rad and QIAGEN digital PCR platforms provide equivalent quantification for wastewater-based SARS-CoV-2 surveillance.Applied and environmental microbiology · 2026Article
- Development and Validation of a Quantitative RT-qPCR Panel for the Detection and Monitoring of Polioviruses in Wastewater Samples.Microorganisms · 2026Article
- Environmental and microbial factors shaping SARS-CoV-2 RNA decay in wastewater: insights from batch tests and a lab-scale sewer pipeline simulator.Scientific reports · 2026Article
- Waterborne pathogen mitigation: Decoding techno-ecological synergies in multiscale transmission networks.Innovation (Cambridge (Mass.)) · 2026Review
- Towards ultra-sensitive and rapid near-source wastewater-based epidemiology.Nature communications · 2025Article
- Comparative evaluation of heating instruments for detectingJournal of water and health · 2025Article
- An integrated methodological framework for the validation and verification of clinical testing by qRT-PCR.Heliyon · 2025Article
- Environmental Dissemination of SARS-CoV-2: An Analysis Employing Crassphage and Next-Generation Sequencing Protocols.Food and environmental virology · 2025Article
- Digital PCR in Virology: Current Applications and Future Perspectives.Molecular diagnosis & therapy · 2025Review
- Surveillance of SARS-CoV-2 in wastewater by quantitative PCR and digital PCR: a case study in Shijiazhuang city, Hebei province, China.Emerging microbes & infections · 2024Article
- A Narrative Review of High Throughput Wastewater Sample Processing for Infectious Disease Surveillance: Challenges, Progress, and Future Opportunities.International journal of environmental research and public health · 2024Review
- Comparison of Different Reverse Transcriptase-Polymerase Chain Reaction-Based Methods for Wastewater Surveillance of SARS-CoV-2: Exploratory Study.JMIR public health and surveillance · 2024Article
- Evaluation of the Impact of Concentration and Extraction Methods on the Targeted Sequencing of Human Viruses from Wastewater.Environmental science & technology · 2024Article
- Wastewater-based surveillance as a tool for public health action: SARS-CoV-2 and beyond.Clinical microbiology reviews · 2024Review
- Optimal Selection of Sampling Points within Sewer Networks for Wastewater-Based Epidemiology Applications.Methods and protocols · 2024Article
- Underestimation of SARS-CoV-2 in wastewater due to single or double mutations in the N1 qPCR probe binding region.Water research X · 2024Article
- Assessment of virus concentration methods for detecting SARS-CoV-2 IN wastewater.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2023Article
- Occurrence of multiple respiratory viruses in wastewater in Queensland, Australia: Potential for community disease surveillance.The Science of the total environment · 2023Article
- Wastewater-based prediction of COVID-19 cases using a highly sensitive SARS-CoV-2 RNA detection method combined with mathematical modeling.Environment international · 2023Article
Corrections and comments
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Authors and funding
7 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Effective wastewater surveillance of SARS-CoV-2 RNA requires the rigorous characterization of the limit of detection resulting from the entire sampling process - the process limit of detection (PLOD). Yet to date, no studies have gone beyond quantifying the assay limit of detection (ALOD) for RT-qPCR or RT-dPCR assays. While the ALOD is the lowest number of gene copies (GC) associated with a 95% probability of detection in a single PCR reaction, the PLOD represents the sensitivity of the method after considering the efficiency of all processing steps (e.g., sample handling, concentration, nucleic acid extraction, and PCR assays) to determine the number of GC in the wastewater sample matrix with a specific probability of detection. The primary objective of this study was to estimate the PLOD resulting from the combination of primary concentration and extraction with six SARS-CoV-2 assays: five RT-qPCR assays (US CDC N1 and N2, China CDC N and ORF1ab (CCDC N and CCDC ORF1ab), and E_Sarbeco RT-qPCR, and one RT-dPCR assay (US CDC N1 RT-dPCR) using two models (exponential survival and cumulative Gaussian). An adsorption extraction (AE) concentration method (i.e., virus adsorption on membrane and the RNA extraction from the membrane) was used to concentrate gamma-irradiated SARS-CoV-2 seeded into 36 wastewater samples. Overall, the US CDC N1 RT-dPCR and RT-qPCR assays had the lowest ALODs (< 10 GC/reaction) and PLODs (<3,954 GC/50 mL; 95% probability of detection) regardless of the seeding level and model used. Nevertheless, consistent amplification and detection rates decreased when seeding levels were < 2.32 × 10
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