Bioaerosol sampling for the detection of aerosolized influenza virus, Influenza Other Respir

Centers for Disease Control, National Institute for Occupational Safety and Health, Health Effects Laboratory Division, Allergy and Clinical Immunology Branch, Morgantown, WV 26505, USA.
Influenza and Other Respiratory Viruses (Impact Factor: 2.2). 06/2007; 1(3):113-20. DOI: 10.1111/j.1750-2659.2007.00020.x
Source: PubMed


Influenza virus was used to characterize the efficacy of a cyclone-based, two-stage personal bioaerosol sampler for the collection and size fractionation of aerosolized viral particles.
A Collison single-jet nebulizer was used to aerosolize the attenuated FluMist vaccine into a calm-air settling chamber. Viral particles were captured with bioaerosol samplers that utilize 2 microcentrifuge tubes to collect airborne particulates. The first tube (T1) collects particles greater than 1.8 microm in diameter, while the second tube (T2) collects particles between 1.0 and 1.8 microm, and the back-up filter (F) collects submicron particles. Following aerosolization, quantitative PCR was used to detect and quantify H1N1 and H3N2 influenza strains.
Based on qPCR results, we demonstrate that aerosolized viral particles were efficiently collected and separated according to aerodynamic size using the two-stage bioaerosol sampler. Most viral particles were collected in T2 (1-1.8 microm) and on the back-up filter (< 1 microm) of the bioaerosol sampler. Furthermore, we found that the detection of viral particles with the two-stage sampler was directly proportional to the collection time. Consequently, viral particle counts were significantly greater at 40 minutes in comparison to 5, 10 and 20 minute aerosol collection points.
Due to a lack of empirical data, aerosol transmission of influenza is often questioned. Using FluMist, we demonstrated that a newly developed bioaerosol sampler is able to recover and size fractionate aerosolized viral particles. This sampler should be an important tool for studying viral transmission in clinical settings and may significantly contribute towards understanding the modes of influenza virus transmission.

Download full-text


Available from: Don H Beezhold
  • Source
    • "Oligo.dT- and UNI-12 primers allow preferentially the retrotranscription mRNA and total influenza RNA, respectively. Real time PCR assays were performed to amplify influenza genes M1 or HA genes [41], [42]. For a control, total RNA and mRNA were measured in influenza-infected macrophages treated with ribavirin (kidney donated from Dr. Nubia Boechat, Farmamginhos, Fiocruz). "
    [Show abstract] [Hide abstract]
    ABSTRACT: HIV-1-infected patients co-infected with A(H1N1)pdm09 surprisingly presented benign clinical outcome. The knowledge that HIV-1 changes the host homeostatic equilibrium, which may favor the patient resistance to some co-pathogens, prompted us to investigate whether HIV-1 infection could influence A(H1N1)pdm09 life cycle in vitro. We show here that exposure of A(H1N1)pdm09-infected epithelial cells to HIV-1 viral particles or its gp120 enhanced by 25% the IFITM3 content, resulting in a decrease in influenza replication. This event was dependent on toll-like receptor 2 and 4. Moreover, knockdown of IFITM3 prevented HIV-1 ability to inhibit A(H1N1)pdm09 replication. HIV-1 infection also increased IFITM3 levels in human primary macrophages by almost 100%. Consequently, the arrival of influenza ribonucleoproteins (RNPs) to nucleus of macrophages was inhibited, as evaluated by different approaches. Reduction of influenza RNPs entry into the nucleus tolled A(H1N1)pdm09 life cycle in macrophages earlier than usual, limiting influenza's ability to induce TNF-α. As judged by analysis of the influenza hemagglutin (HA) gene from in vitro experiments and from samples of HIV-1/A(H1N1)pdm09 co-infected individuals, the HIV-1-induced reduction of influenza replication resulted in delayed viral evolution. Our results may provide insights on the mechanisms that may have attenuated the clinical course of Influenza in HIV-1/A(H1N1)pdm09 co-infected patients during the recent influenza form 2009/2010.
    Full-text · Article · Jun 2014 · PLoS ONE
  • Source
    • "Airborne biological particles drawn into the 1 mm diameter nozzle and down the capillary stem, then impacted and penetrated the liquid surface [32]. The adhesion properties involving liquid and airborne particles were exploited by this technology to capture the microorganisms [33]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Avian H5N1 influenza viruses present a challenge in the laboratory environment, as they are difficult to collect from the air due to their small size and relatively low concentration. In an effort to generate effective methods of H5N1 air removal and ensure the safety of laboratory personnel, this study was designed to investigate the characteristics of aerosolized H5N1 produced by laboratory manipulations during research studies. Normal laboratory procedures used to process the influenza virus were carried out independently and the amount of virus polluting the on-site atmosphere was measured. In particular, zootomy, grinding, centrifugation, pipetting, magnetic stirring, egg inoculation, and experimental zoogenetic infection were performed. In addition, common accidents associated with each process were simulated, including breaking glass containers, syringe injection of influenza virus solution, and rupturing of centrifuge tubes. A micro-cluster sampling ambient air pollution collection device was used to collect air samples. The collected viruses were tested for activity by measuring their ability to induce hemagglutination with chicken red blood cells and to propagate in chicken embryos after direct inoculation, the latter being detected by reverse-transcription PCR and HA test. The results showed that the air samples from the normal centrifugal group and the negative-control group were negative, while all other groups were positive for H5N1. Our findings suggest that there are numerous sources of aerosols in laboratory operations involving H5N1. Thus, laboratory personnel should be aware of the exposure risk that accompanies routine procedures involved in H5N1 processing and take proactive measures to prevent accidental infection and decrease the risk of virus aerosol leakage beyond the laboratory.
    Full-text · Article · Aug 2012 · Virology Journal
  • Source
    • "Techniques employing quantitative PCR (qPCR) have been developed for applications such as determination of viral contamination in water samples (Karim et al. 2009). For the determination of the relative contribution of various routes of exposure to influenza virus infection , quantitative assessment of surface contamination and air concentration in the environment can be done with qPCR using calibration with influenza virus vaccine (Blachere et al. 2007) or calibration with influenza matrix gene DNA (Lindsley et al. 2010). These qPCR techniques have been utilized to assess airborne concentrations in healthcare facilities (Blachere et al. 2009) or from cough samples (Lindsley et al. 2010). "
    [Show abstract] [Hide abstract]
    ABSTRACT: The Luminex xTAG(®) respiratory viral panel (RVP) kit simultaneously detects and identifies multiple respiratory viruses including several subtypes of influenza A using a multiplex nucleic acid amplification test assay platform. The emitted fluorescence signal from the RVP assay provides qualitative information on the presence of a particular viral species in respiratory specimens. However, a quantitative assessment is preferred when monitoring environmental samples for respiratory viruses. In this study, we explored the potential use of the RVP kit as a semi-quantitative screening assay for influenza virus detection. The concentration- response of the RVP assay was modeled using four-parameter logistic (4-PL) fits of mean fluorescence intensity (MFI) versus dilute ranges of the influenza A matrix gene, seasonal influenza vaccine, and 2009 H1N1 influenza vaccine. The goodness of fit of the 4-PL model was evaluated by comparing the copy number determined with the fitted model (observed copy number) with the copy number calculated from the dilution of the matrix DNA or vaccine (expected copy number). For the matrix DNA and 2009 H1N1 vaccine, the 4-PL model provided good fit for the influenza A RVP assay response over factors of 10(3) to 10(4). For seasonal influenza vaccine, the model provided good fit for RVP assay response to influenza A, influenza B, H1, and H3.
    Full-text · Article · Nov 2011 · Toxicology mechanisms and methods
Show more