High air flow-rate electrostatic sampler for the rapid monitoring of airborne coronavirus and influenza viruses.

Capturing virus aerosols in a small volume of liquid is essential when monitoring airborne viruses. As such, aerosol-to-hydrosol enrichment is required to produce a detectable viral sample for real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) assays. To meet this requirement, the efficient and non-destructive collection of airborne virus particles is needed, while the incoming air flow rate should be sufficiently high to quickly collect a large number of virus particles. To achieve this, we introduced a high air flow-rate electrostatic sampler (HAFES) that collected virus aerosols (human coronavirus 229E, influenza A virus subtypes H1N1 and H3N2, and bacteriophage MS2) in a continuously flowing liquid. Viral collection efficiency was evaluated using aerosol particle counts, while viral recovery rates were assessed using real-time qRT-PCR and plaque assays. An air sampling period of 20 min was sufficient to produce a sample suitable for use in real-time qRT-PCR in a viral epidemic scenario.

Aerosol-to-Hydrosol Sampling and Simultaneous Enrichment of Airborne Bacteria For Rapid Biosensing.

Rapid monitoring of biological particulate matter (Bio-PM, bioaerosols) requires an enrichment technique for concentrating the Bio-PM dispersed in the air into a small volume of liquid. In this study, an electrostatic air sampler is employed to capture aerosolized test bacteria in a carrier liquid (aerosol-to-hydrosol (ATH) enrichment). Simultaneously, the captured bacteria are carried into a fluid channel for hydrosol-to-hydrosol (HTH) enrichment with Concanavalin A coated magnetic particles (CMPs). The ATH enrichment capacity of the air sampler was evaluated with an aerosol particle counter for the following test bacteria: Staphylococcus aureus, Bacillus cereus, Escherichia coli, and Acinetobacter baumannii. Then, the HTH enrichment capacity for the ATH-collected sample was evaluated using the colony-counting method, scanning electron microscopy based image analysis, fluorescence microscopy, electrical current measurements, and real-time quantitative polymerase chain reaction (qPCR). The ATH and HTH enrichment capacities for the given operation conditions were up to 80 000 and 14.9, respectively, resulting in a total enrichment capacity of up to 1.192 × 106. Given that air-to-liquid enrichment required to prepare detectable bacterial samples for real-time qPCR in field environments is of the order of at least 106, our method can be used to prepare a detectable sample from low-concentration airborne bacteria in the field and significantly reduce the time required for Bio-PM monitoring because of its enrichment capacity.

An integrated system of air sampling and simultaneous enrichment for rapid biosensing of airborne coronavirus and influenza virus.

Point-of-care risk assessment (PCRA) for airborne viruses requires a system that can enrich low-concentration airborne viruses dispersed in field environments into a small volume of liquid. In this study, airborne virus particles were collected to a degree above the limit of detection (LOD) for a real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). This study employed an electrostatic air sampler to capture aerosolized test viruses (human coronavirus 229E (HCoV-229E), influenza A virus subtype H1N1 (A/H1N1), and influenza A virus subtype H3N2 (A/H3N2)) in a continuously flowing liquid (aerosol-to-hydrosol (ATH) enrichment) and a concanavalin A (ConA)-coated magnetic particles (CMPs)-installed fluidic channel for simultaneous hydrosol-to-hydrosol (HTH) enrichment. The air sampler's ATH enrichment capacity (EC) was evaluated using the aerosol counting method. In contrast, the HTH EC for the ATH-collected sample was evaluated using transmission-electron-microscopy (TEM)-based image analysis and real-time qRT-PCR assay. For example, the ATH EC for HCoV-229E was up to 67,000, resulting in a viral concentration of 0.08 PFU/mL (in a liquid sample) for a viral epidemic scenario of 1.2 PFU/m3 (in air). The real-time qRT-PCR assay result for this liquid sample was "non-detectable" however, subsequent HTH enrichment for 10 min caused the "non-detectable" sample to become "detectable" (cycle threshold (CT) value of 33.8 ± 0.06).

The Gillings Sampler--an electrostatic air sampler as an alternative method for aerosol in vitro exposure studies.

There is growing interest in studying the toxicity and health risk of exposure to multi-pollutant mixtures found in ambient air, and the U.S. Environmental Protection Agency (EPA) is moving towards setting standards for these types of mixtures. Additionally, the Health Effects Institute's strategic plan aims to develop and apply next-generation multi-pollutant approaches to understanding the health effects of air pollutants. There's increasing concern that conventional in vitro exposure methods are not adequate to meet EPA's strategic plan to demonstrate a direct link between air pollution and health effects. To meet the demand for new in vitro technology that better represents direct air-to-cell inhalation exposures, a new system that exposes cells at the air-liquid interface was developed. This new system, named the Gillings Sampler, is a modified two-stage electrostatic precipitator that provides a viable environment for cultured cells. Polystyrene latex spheres were used to determine deposition efficiencies (38-45%), while microscopy and imaging techniques were used to confirm uniform particle deposition. Negative control A549 cell exposures indicated the sampler can be operated for up to 4h without inducing any significant toxic effects on cells, as measured by lactate dehydrogenase (LDH) and interleukin-8 (IL-8). A novel positive aerosol control exposure method, consisting of a p-tolualdehyde (TOLALD) impregnated mineral oil aerosol (MOA), was developed to test this system. Exposures to the toxic MOA at a 1 ng/cm(2) dose of TOLALD yielded a reproducible 1.4 and 2-fold increase in LDH and IL-8 mRNA levels over controls. This new system is intended to be used as an alternative research tool for aerosol in vitro exposure studies. While further testing and optimization is still required to produce a "commercially ready" system, it serves as a stepping-stone in the development of cost-effective in vitro technology that can be made accessible to researchers in the near future.