Kean, Michael

Caffrey, Paul

doi:10.3318/dib.004395.v1

Cited by 1 publication

(3 citation statements)

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“…Because we are conducting the NAA reaction directly in the collection well, we can circumvent the usual extraction losses from viral extraction off the collection surface using an extraction buffer and the subsequent concentration of viral RNA within the extract solution. We anticipate therefore that the number of viral copies per well for the one-pot reaction is close to the sensitivity of the RT-LAMP assay i.e., less than 30 RNA copies [42,49], a result we have confirmed experimentally using stock VLP solution. This will be further analyzed using aerosolized VLPs in (ongoing) future work.…”

Section: Discussionsupporting

confidence: 61%

“…Once charged, electrohydrodynamic transport of charged aerosol droplets causes their precipitation onto the grounded collection electrode surface [30]. While low collection efficiency is expected for such a high flow rate unoptimized collector, the on-chip integration of a highly sensitive detection technique (RT-LAMP) means that virions collected directly at the collection site can be detected at very low numbers (~ 30 RNA copies) [42,49]. Longer sampling times are needed when airborne viral concentration is low.…”

Section: Electrostatic Viral Aerosol Precipitationmentioning

confidence: 99%

“…As detailed in [42] we developed a BSL2 model for airborne viruses using VLP containing the SARS-CoV-2 spike protein in the membrane. In this model, the VLP contains a copy of the HIV genome in the form of RNA, which includes the HIV pol gene.…”

Section: Experimental Characterization Of Vlp Aerosolizationmentioning

confidence: 99%

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Towards Real-Time Airborne Pathogen Sensing: Electrostatic Capture and On-Chip LAMP Based Detection of Airborne Viral Pathogens

Jayakumar,

Caffrey,

Caffrey

et al. 2024

Preprint

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Considerable loss of life, economic slowdown, and public health risk associated with the transmission of airborne respiratory pathogens was underscored by the recent COVID-19 pandemic. Airborne transmission of zoonotic diseases such as the highly pathogenic avian influenza (HPAI) and porcine reproductive and respiratory syndrome virus (PRRSV) has caused major disruptions to domestic and global food security. Current ambient air pathogen monitoring systems involves the collection of air samples from indoor settings suspected of viral contamination, followed by subsequent processing of capture samples to determine the presence and species of airborne viral matter. Nucleic acid amplification techniques are considered the gold standard for pathogen diagnostics. Currently, the necessary extraction and purification of viral RNA from air collector systems prior to sample analysis is both time consuming and performed manually. A monitoring system with separate air sampling and biochemical detection procedures is prone to delay the response to emergent viral threats. In this paper, we present a pathogen monitoring system that overcomes these limitations related to extraction and purification of viral samples and lays the groundwork for a real-time monitor for airborne viral pathogens. We demonstrate a high flow electrostatic precipitator system, that uses small collection wells as counter electrodes for pathogen collection. Integrated reverse-transcriptase loop-mediated isothermal amplification (RT-LAMP) is used for detection of captured viral matter within wells. On-chip heating of collection wells is enabled by integrated planar heaters and small volumes of reagent (30 μL) directly to the collection wells. We present the design of such a system and show experimental results that demonstrate the use of this device for detection of aerosolized SARS-CoV-2 virus like particles (VLPs), a model pathogen for SARV-CoV-2.

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Section: Discussionsupporting

confidence: 61%