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Characterization of a fieldable process for airborne virus detection

Du, H.; Bruno, S.; Overholt, K. J.; Palacios, S.; Huang, H.-H.; Barajas, C.; Gross, B.; Lee, C.; Evile, H. K.; de Sousa, N. R.; Rothfuchs, A. G.; Del Vecchio, D.

2023-07-06 infectious diseases
10.1101/2023.07.03.23292170 medRxiv
Show abstract

Rapid, on-site, airborne virus detection is a requirement for timely action against the spread of air-transmissible infectious diseases. This applies both to future threats and to common viral diseases, such as influenza and COVID-19, which hit vulnerable populations yearly with severe consequences. The ultra-low concentrations of virus in the air make airborne virus detection difficult, yet readily infect individuals when breathed. Here, we propose a fieldable process that includes an enrichment step to concentrate collected genetic material in a small volume. The enrichment approach uses capillary electrophoresis and an RT-qPCR-compatible buffer, which allow enrichment of the RNA by about 5-fold within only 10 minutes of operation. Our detection process consists of air sampling through electrostatic precipitation, RNA extraction via heating, RNA enrichment, and RT-qPCR for detection. We optimized each step of the process and estimated a detection sensitivity of 3106 {+/-} 2457 genome copies (gc) per m3 of air. We then performed an integration experiment and confirmed a sensitivity of 5654 gc/m3 with a detection rate of 100% and a sensitivity of 4221 gc/m3 with a detection rate of 78.6%. When using fast RT-qPCR, the latency of the whole process is down to 61 minutes. Given that our sensitivity falls in the low range of influenza and SARS-CoV-2 concentrations reported in indoor spaces, our study shows that, with enrichment, airborne pathogen detection can be made sufficiently sensitive for practical use.

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