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Platinum chloride-based viability RT-qPCR for SARS-CoV-2 detection in complex samples.
Enric Cuevas-Ferrando1, Walter Randazzo2, Alba Pérez-Cataluña2
1Department of Preservation and Food Safety Technologies, Institute of Agrochemistry and Food Technology, IATA-CSIC, Av. Agustín Escardino 7, 46980, Paterna, Valencia, Spain. enric.cuevas@iata.csic.es.
Scientific Reports
|September 14, 2021
Summary
A new viability RT-qPCR method accurately detects infectious SARS-CoV-2, distinguishing viable virus from RNA in clinical and environmental samples. This improves infection risk assessment for controlling outbreaks.
Area of Science:
- Virology
- Molecular Biology
- Public Health
Background:
- Current SARS-CoV-2 detection methods (RT-qPCR) detect RNA but not infectivity, potentially overestimating infection risk.
- Global control measures like isolation and contact tracing rely on accurate assessment of infectious virus presence.
Purpose of the Study:
- To develop a rapid viability RT-qPCR assay to differentiate between infectious and non-infectious SARS-CoV-2 particles.
- To assess the infectivity of SARS-CoV-2 in clinical specimens and environmental samples.
Main Methods:
- Screening of viability markers (monoazide dyes, platinum compounds) against SARS-CoV-2 RNA targets.
- Optimization of a platinum chloride-based viability RT-qPCR using genomic RNA and inactivated SARS-CoV-2 in various matrices (buffer, stool, urine).
- Validation in COVID-19 positive nasopharyngeal swabs and SARS-CoV-2 RNA positive wastewater.
Main Results:
- A rapid viability RT-qPCR was successfully established, selectively detecting potentially infectious SARS-CoV-2.
- Nasopharyngeal swabs from COVID-19 patients showed positivity, indicating potential infectivity.
- Wastewater samples showed no amplification, suggesting non-infectious particles or free RNA.
Conclusions:
- The viability RT-qPCR offers a more accurate method for detecting infectious SARS-CoV-2.
- This approach can enhance risk-based investigations for preventing and controlling outbreaks.
- The assay has broad applicability for various sample types.
