ArticleThe Science of the total environment2021
Rapid determination of infectious SARS-CoV-2 in PCR-positive samples by SDS-PMA assisted RT-qPCR.
Article in The Science of the total environment, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 35 citations in OpenAlex.
- From clinical fluids to environmental matrices: quantitative monitoring of infectious monkeypox virus based on PMA-qPCR during the Guangdong outbreak.Tropical medicine and health · 2026Article
- Molecular Differentiation of Intact West Nile Virus Using a PMAxx™-Enabled Digital PCR Workflow.International journal of molecular sciences · 2026Article
- Earlier de-isolation of SARS-CoV-2-infected ICU patients using a novel viability PCR: a prospective cohort study.Microbiology spectrum · 2026Article
- Rapid differentiation of viable and inactivated African swine fever virus by a viability quantitative PCR.Veterinary research · 2025Article
- Rapid and accurate detection of infectious SARS-CoV-2 by viral receptor capture combined with loop-mediated isothermal amplification.Virologica Sinica · 2025Article
- EnhancingMicrobiology spectrum · 2025Article
- Measurement of the Infection and Integrity of Monkeypox Virus: A New Method Using PMAxx-ddPCR.International journal of molecular sciences · 2025Article
- A Triplex Propidium Monoazide (PMA) qPCR Assay Enables Rapid Discrimination of Live Porcine Reproductive and Respiratory Syndrome Viruses.Transboundary and emerging diseases · 2025Article
- Rapidly quantification of intact infectious H1N1 virus using ICA-qPCR and PMA-qPCR.Biosafety and health · 2024Article
- Advancing COVID-19 diagnostics: rapid detection of intact SARS-CoV-2 using viability RT-PCR assay.Microbiology spectrum · 2024Article
- Sodium Deoxycholate-Propidium Monoazide Droplet Digital PCR for Rapid and Quantitative Detection of ViableMicroorganisms · 2024Article
- Rapid Determination of SARS-CoV-2 Integrity and Infectivity by Using Propidium Monoazide Coupled with Digital Droplet PCR.International journal of molecular sciences · 2024Article
- Improving the Efficiency of Viability-qPCR with Lactic Acid Enhancer for the Selective Detection of Live Pathogens in Foods.Foods (Basel, Switzerland) · 2024Article
- Propidium Monoazide Integrated With qPCR Enables Rapid and Universal Detection of Infectious Porcine Reproductive and Respiratory Syndrome Viruses.Transboundary and emerging diseases · 2024Article
- Ultrafast inactivation of SARS-CoV-2 by 254-nm UV-C irradiation on porous and non-porous media of medical interest using an omnidirectional chamber.Scientific reports · 2023Article
- Article
- Review
- Digital PCR Applications in the SARS-CoV-2/COVID-19 Era: a Roadmap for Future Outbreaks.Clinical microbiology reviews · 2022Review
- Rapid Quantification of InfectiousViruses · 2022Article
- Quantification of infectious Human mastadenovirus in environmental matrices using PMAxx-qPCR.Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] · 2022Article
Corrections and comments
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Authors and funding
9 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The ongoing COVID-19 pandemic has generated a global health crisis that needs well management of not only patients but also environments to reduce SARS-CoV-2 transmission. The gold standard RT-qPCR method is sensitive and rapid to detect SARS-CoV-2 nucleic acid, but does not answer if PCR-positive samples contain infectious virions. To circumvent this problem, we report an SDS-propidium monoazide (PMA) assisted RT-qPCR method that enables rapid discrimination of live and dead SARS-CoV-2 within 3 h. PMA, a photo-reactive dye, can react with viral RNA released or inside inactivated SARS-CoV-2 virions under assistance of 0.005% SDS, but not viral RNA inside live virions. Formation of PMA-RNA conjugates prevents PCR amplification, leaving only infectious virions to be detected. Under optimum conditions, RT-qPCR detection of heat-inactivated SARS-CoV-2 resulted in larger than 9 Ct value differences between PMA-treated and PMA-free groups, while less than 0.5 Ct differences were observed in the detection of infectious SARS-CoV-2 ranging from 20 to 5148 viral particles. Using a cutoff Ct difference of 8.6, this method could differentiate as low as 8 PFU live viruses in the mixtures of live and heat-inactivated virions. Further experiments showed that this method could successfully monitor the natural inactivation process of SARS-CoV-2 on plastic surfaces during storage with comparable results to the gold standard plaque assay. We believe that the culture-free method established here could be used for rapid and convenient determination of infectious SARS-CoV-2 virions in PCR-positive samples, which will facilitate better control of SARS-CoV-2 transmission.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.