Integration of sample preparation with RNA-Amplification in a hand-held device for airborne virus detection

Xiao Jiang; Julia C Loeb; Maohua Pan; Trevor B Tilly; Arantza Eiguren-Fernandez; John A Lednicky; Chang-Yu Wu; Z Hugh Fan

doi:10.1016/j.aca.2021.338542

Integration of sample preparation with RNA-Amplification in a hand-held device for airborne virus detection

Anal Chim Acta. 2021 Jun 22:1165:338542. doi: 10.1016/j.aca.2021.338542. Epub 2021 Apr 23.

Authors

Xiao Jiang¹, Julia C Loeb², Maohua Pan³, Trevor B Tilly³, Arantza Eiguren-Fernandez⁴, John A Lednicky⁵, Chang-Yu Wu⁶, Z Hugh Fan⁷

Affiliations

¹ J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, P.O. Box 116131, Gainesville, FL, 32611, USA.
² Department of Environmental and Global Health, and Emerging Pathogens Institute, University of Florida, Gainesville, FL, USA.
³ Department of Environmental Engineering Sciences, Engineering School of Sustainable Infrastructure and Environment, University of Florida, Gainesville, FL, USA.
⁴ Aerosol Dynamics Inc., Berkeley, CA, USA.
⁵ Department of Environmental and Global Health, and Emerging Pathogens Institute, University of Florida, Gainesville, FL, USA. Electronic address: jlednicky@phhp.ufl.edu.
⁶ Department of Environmental Engineering Sciences, Engineering School of Sustainable Infrastructure and Environment, University of Florida, Gainesville, FL, USA. Electronic address: cywu@ufl.edu.
⁷ J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, P.O. Box 116131, Gainesville, FL, 32611, USA; Department of Mechanical and Aerospace Engineering, University of Florida, P.O. Box 116250, Gainesville, FL, 32611, USA; Department of Chemistry, University of Florida, P.O. Box 117200, Gainesville, FL, 32611, USA. Electronic address: hfan@ufl.edu.

Abstract

Aerosol transmission is one of the three major transmission routes of respiratory viruses. However, the dynamics and significance of the aerosol transmission route are not well understood, partially due to the lack of rapid and efficient tools for on-the-spot detection of airborne viruses. We report a hand-held device that integrates a 3D-printed sample preparation unit with a laminated paper-based RNA amplification unit. The sample preparation unit features an innovative reagent delivery scheme based on a ball-based valve capable of storing and delivering reagents through the rotation of the unit without manual pipetting, while the paper-based unit enables RNA enrichment and reverse transcription loop-mediated isothermal amplification (RT-LAMP). We have determined the detection limit of the integrated sample-preparation/amplification device (SPAD) at 1 TCID₅₀ H1N1 influenza viruses in 140 μL aqueous sample. Further, we integrated SPAD with a previously reported viable virus aerosol sampler (VIVAS), a water-vapor-based condensational growth system capable of collecting aerosolized virus particles (Pan et al., 2016) [1]. Using the combined VIVAS-SPAD platform, we have demonstrated the collection/detection of lab-generated, airborne H1N1 influenza viruses in 65 min, suggesting that the platform has a potential for detecting and monitoring airborne virus transmission during outbreaks. The effective sampling and rapid detection of airborne viruses by the sample-to-answer platform will also help us better understand the dynamics and significance of aerosol transmission of infectious disease.

Keywords: Airborne; Influenza virus; Isothermal amplification; Paper microfluidics; Sample preparation; Valve.

MeSH terms

Influenza A Virus, H1N1 Subtype* / genetics
Influenza A virus* / genetics
Molecular Diagnostic Techniques
Nucleic Acid Amplification Techniques
RNA

Substances

RNA

Supplementary concepts

LAMP assay

Grants and funding

R01 AI158868/AI/NIAID NIH HHS/United States