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Efficiency and sensitivity optimization of a protocol to quantify indoor airborne SARS-CoV-2 levels
J Truyols-Vives1, K Stiliyanov-Atanasov1, E Sala-Llinàs2
1Molecular Biology and One Health Research Group (MolONE), Universitat de les Illes Balears (UIB), Palma, Spain.
The Journal of Hospital Infection
|September 13, 2022
Summary
Optimized air sampling using mineral oil and SuperScript II reverse transcriptase (RT) enhances detection of airborne SARS-CoV-2 RNA. This improved method increases sensitivity for assessing indoor air microbiological safety.
Area of Science:
- Environmental microbiology
- Molecular diagnostics
- Infectious disease control
Background:
- Quantifying airborne microorganisms is crucial for infection prevention.
- Assessing airborne SARS-CoV-2 RNA levels lacks a consistently efficient and sensitive strategy.
- Previous protocols for SARS-CoV-2 RNA measurement require improvement.
Purpose of the Study:
- To enhance a previously established protocol for measuring SARS-CoV-2 RNA in bioaerosols.
- To optimize methods for collecting and quantifying airborne SARS-CoV-2 RNA.
- To improve the sensitivity and efficiency of detecting SARS-CoV-2 RNA in indoor air.
Main Methods:
- Air samples were collected from COVID-19 patient rooms.
- Evaluated different air samplers, liquid collection media (mineral oil vs. hydrophilic), and reverse transcriptases (RT).
- Quantified SARS-CoV-2 RNA levels using droplet digital polymerase chain reaction (ddPCR).
Main Results:
- Mineral oil demonstrated superior airborne RNA retention compared to hydrophilic media.
- The BioSampler with mineral oil detected SARS-CoV-2 ORF1ab target in 80% of samples.
- SuperScript II RT enabled high-efficiency detection of single SARS-CoV-2 genome RNA molecules and was essential for detecting the N1 target.
Conclusions:
- The study successfully improved the collection efficiency and detection sensitivity of a protocol for quantifying airborne SARS-CoV-2 RNA.
- Optimized methods enhance the understanding of microbiological safety in indoor air environments.
- The refined protocol offers a more sensitive approach to monitoring airborne viral RNA.
