ArticleHuman genomics2024
Simultaneous detection of influenza A, B and respiratory syncytial virus in wastewater samples by one-step multiplex RT-ddPCR assay.
Article in Human genomics, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed.
- Air Purifier Intervention for Respiratory Viral Exposure in Elementary Schools: A Secondary Analysis of a Randomized Clinical Trial.JAMA network open · 2025Trial
- Detection of Respiratory Viruses in Wastewater Samples Using Commercial Multiplex PCR Assays.Viruses · 2026Article
- Real-time estimation of pathogen transmission dynamics from wastewater.Nature communications · 2026Article
- Article
- RAZOR: a database of PCR primers targeting human respiratory viruses.NAR genomics and bioinformatics · 2026Article
- One Assay, Nine Targets: Advancing Viral Surveillance with Multiplex RT-ddPCR.Analytical chemistry · 2025Article
- Development and validation of an automated and high-throughput quadruplex RT-ddPCR assay for the detection of influenza A, influenza B, respiratory syncytial virus, and SARS-CoV-2.Frontiers in cellular and infection microbiology · 2025Article
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Authors and funding
4 authors.
Funding
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Abstract
backgroundAfter the occurrence of the COVID-19 pandemic, detection of other disseminated respiratory viruses using highly sensitive molecular methods was declared essential for monitoring the spread of health-threatening viruses in communities. The development of multiplex molecular assays are essential for the simultaneous detection of such viruses even at low concentrations. In the present study, a highly sensitive and specific multiplex one-step droplet digital PCR (RT-ddPCR) assay was developed for the simultaneous detection and absolute quantification of influenza A (IAV), influenza B (IBV), respiratory syncytial virus (RSV), and beta-2-microglobulin transcript as an endogenous internal control (IC B2M).
resultsThe assay was first evaluated for analytical sensitivity and specificity, linearity, reproducibility, and recovery rates with excellent performance characteristics and then applied to 37 wastewater samples previously evaluated with commercially available and in-house quantitative real-time reverse transcription PCR (RT-qPCR) assays. IAV was detected in 16/37 (43%), IBV in 19/37 (51%), and RSV in 10/37 (27%) of the wastewater samples. Direct comparison of the developed assay with real-time RT-qPCR assays showed statistically significant high agreement in the detection of IAV (kappa Cohen's correlation coefficient: 0.834, p = 0.001) and RSV (kappa: 0.773, p = 0.001) viruses between the two assays, while the results for the detection of IBV (kappa: 0.355, p = 0.27) showed good agreement without statistical significance.
conclusionsOverall, the developed one-step multiplex ddPCR assay is cost-effective, highly sensitive and specific, and can simultaneously detect three common respiratory viruses in the complex matrix of wastewater samples even at low concentrations. Due to its high sensitivity and resistance to PCR inhibitors, the developed assay could be further used as an early warning system for wastewater monitoring.
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