Capture and visualization of live Mycobacterium tuberculosis bacilli from tuberculosis patient bioaerosols

Autor: Robin Wood, Andrea McKerry, Bryan Leonard, Anastasia Koch, Mireille Kamariza, Sophia Gessner, Ronnett Seldon, Ryan Dinkele, Andrew Kamholz, Carolyn R. Bertozzi, Gilla Kaplan, Valerie Mizrahi, Wayne A. Bryden, Courtney McLoud, Charles Call, Brian J. Smith, Digby F. Warner, Melitta Gqada, Carl Morrow
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Bacterial Diseases
Pulmonology
Pathology and Laboratory Medicine
Cohort Studies
0302 clinical medicine
Medical Conditions
Fluorescence Microscopy
Medicine and Health Sciences
Nanotechnology
030212 general & internal medicine
Biology (General)
Materials
Aerosolization
0303 health sciences
Microscopy
Transmission (medicine)
Metabolic heterogeneity
Light Microscopy
Bacterial Pathogens
Actinobacteria
Infectious Diseases
Breath Tests
Medical Microbiology
Physical Sciences
Engineering and Technology
Pathogens
Bioaerosol
Research Article
Biotechnology
Adult
Bacilli
Tuberculosis
QH301-705.5
Imaging Techniques
Immunology
Indoor bioaerosol
Materials Science
Bioengineering
Biology
Research and Analysis Methods
Microbiology
Mycobacterium tuberculosis
03 medical and health sciences
Respiratory Disorders
Virology
Fluorescence Imaging
Genetics
medicine
Humans
Molecular Biology
Microbial Pathogens
Tuberculosis
Pulmonary
030304 developmental biology
Aerosols
Bacteria
Organisms
Biology and Life Sciences
Mycobacteria
RC581-607
medicine.disease
biology.organism_classification
bacterial infections and mycoses
Tropical Diseases
Cough
Microscopy
Fluorescence
Mixtures
Bionanotechnology
Respiratory Infections
Parasitology
Immunologic diseases. Allergy
Zdroj: PLoS Pathogens
PLoS Pathogens, Vol 17, Iss 2, p e1009262 (2021)
ISSN: 1553-7374
1553-7366
Popis: Interrupting transmission is an attractive anti-tuberculosis (TB) strategy but it remains underexplored owing to our poor understanding of the events surrounding transfer of Mycobacterium tuberculosis (Mtb) between hosts. Determining when live, infectious Mtb bacilli are released and by whom has proven especially challenging. Consequently, transmission chains are inferred only retrospectively, when new cases are diagnosed. This process, which relies on molecular analyses of Mtb isolates for epidemiological fingerprinting, is confounded by the prolonged infectious period of TB and the potential for transmission from transient exposures. We developed a Respiratory Aerosol Sampling Chamber (RASC) equipped with high-efficiency filtration and sampling technologies for liquid-capture of all particulate matter (including Mtb) released during respiration and non-induced cough. Combining the mycobacterial cell wall probe, DMN-trehalose, with fluorescence microscopy of RASC-captured bioaerosols, we detected and quantified putative live Mtb bacilli in bioaerosol samples arrayed in nanowell devices. The RASC enabled non-invasive capture and isolation of viable Mtb from bioaerosol within 24 hours of collection. A median 14 live Mtb bacilli (range 0–36) were isolated in single-cell format from 90% of confirmed TB patients following 60 minutes bioaerosol sampling. This represented a significant increase over previous estimates of transmission potential, implying that many more organisms might be released daily than commonly assumed. Moreover, variations in DMN-trehalose incorporation profiles suggested metabolic heterogeneity in aerosolized Mtb. Finally, preliminary analyses indicated the capacity for serial image capture and analysis of nanowell-arrayed bacilli for periods extending into weeks. These observations support the application of this technology to longstanding questions in TB transmission including the propensity for asymptomatic transmission, the impact of TB treatment on Mtb bioaerosol release, and the physiological state of aerosolized bacilli.
Author summary Mycobacterium tuberculosis (Mtb), the cause of tuberculosis (TB), must drive successive cycles of transmission and infection to retain a foothold in its obligate human host. Although critical for Mtb survival, the mechanisms enabling successful transmission have largely evaded research owing to the difficulties inherent in identifying when bacilli are released and by whom. With the available tools, fewer than one-third of new Mtb infections can be confidently linked to known TB cases, a deficiency reflecting the confounding effects of the prolonged infectious period of TB and the potential for transmission from transient exposures. Here, we describe the deployment of the Respiratory Aerosol Sampling Chamber (RASC), a personal clean room equipped for high-efficiency capture of bioaerosols, to isolate live Mtb bacilli released in infectious aerosols. Applying a fluorescent viability probe and microscopic imaging, we demonstrate the detection of live Mtb with single-cell resolution in complex bioaerosol samples from a high proportion of TB cases. Moreover, by exploiting compartmentalization of bacilli within a nanowell collection device, we establish the capacity for long-term maintenance of bacillary viability for serial imaging. Our observations support the utility of the RASC to better understand and ultimately interrupt Mtb transmission.
Databáze: OpenAIRE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje