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

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 pro...

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Autores principales: Dinkele, Ryan, Gessner, Sophia, McKerry, Andrea, Leonard, Bryan, Seldon, Ronnett, Koch, Anastasia S., Morrow, Carl, Gqada, Melitta, Kamariza, Mireille, Bertozzi, Carolyn R., Smith, Brian, McLoud, Courtney, Kamholz, Andrew, Bryden, Wayne, Call, Charles, Kaplan, Gilla, Mizrahi, Valerie, Wood, Robin, Warner, Digby F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7877778/
https://www.ncbi.nlm.nih.gov/pubmed/33524021
http://dx.doi.org/10.1371/journal.ppat.1009262
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author Dinkele, Ryan
Gessner, Sophia
McKerry, Andrea
Leonard, Bryan
Seldon, Ronnett
Koch, Anastasia S.
Morrow, Carl
Gqada, Melitta
Kamariza, Mireille
Bertozzi, Carolyn R.
Smith, Brian
McLoud, Courtney
Kamholz, Andrew
Bryden, Wayne
Call, Charles
Kaplan, Gilla
Mizrahi, Valerie
Wood, Robin
Warner, Digby F.
author_facet Dinkele, Ryan
Gessner, Sophia
McKerry, Andrea
Leonard, Bryan
Seldon, Ronnett
Koch, Anastasia S.
Morrow, Carl
Gqada, Melitta
Kamariza, Mireille
Bertozzi, Carolyn R.
Smith, Brian
McLoud, Courtney
Kamholz, Andrew
Bryden, Wayne
Call, Charles
Kaplan, Gilla
Mizrahi, Valerie
Wood, Robin
Warner, Digby F.
author_sort Dinkele, Ryan
collection PubMed
description 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.
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spelling pubmed-78777782021-02-19 Capture and visualization of live Mycobacterium tuberculosis bacilli from tuberculosis patient bioaerosols Dinkele, Ryan Gessner, Sophia McKerry, Andrea Leonard, Bryan Seldon, Ronnett Koch, Anastasia S. Morrow, Carl Gqada, Melitta Kamariza, Mireille Bertozzi, Carolyn R. Smith, Brian McLoud, Courtney Kamholz, Andrew Bryden, Wayne Call, Charles Kaplan, Gilla Mizrahi, Valerie Wood, Robin Warner, Digby F. PLoS Pathog Research Article 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. Public Library of Science 2021-02-01 /pmc/articles/PMC7877778/ /pubmed/33524021 http://dx.doi.org/10.1371/journal.ppat.1009262 Text en © 2021 Dinkele et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Dinkele, Ryan
Gessner, Sophia
McKerry, Andrea
Leonard, Bryan
Seldon, Ronnett
Koch, Anastasia S.
Morrow, Carl
Gqada, Melitta
Kamariza, Mireille
Bertozzi, Carolyn R.
Smith, Brian
McLoud, Courtney
Kamholz, Andrew
Bryden, Wayne
Call, Charles
Kaplan, Gilla
Mizrahi, Valerie
Wood, Robin
Warner, Digby F.
Capture and visualization of live Mycobacterium tuberculosis bacilli from tuberculosis patient bioaerosols
title Capture and visualization of live Mycobacterium tuberculosis bacilli from tuberculosis patient bioaerosols
title_full Capture and visualization of live Mycobacterium tuberculosis bacilli from tuberculosis patient bioaerosols
title_fullStr Capture and visualization of live Mycobacterium tuberculosis bacilli from tuberculosis patient bioaerosols
title_full_unstemmed Capture and visualization of live Mycobacterium tuberculosis bacilli from tuberculosis patient bioaerosols
title_short Capture and visualization of live Mycobacterium tuberculosis bacilli from tuberculosis patient bioaerosols
title_sort capture and visualization of live mycobacterium tuberculosis bacilli from tuberculosis patient bioaerosols
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7877778/
https://www.ncbi.nlm.nih.gov/pubmed/33524021
http://dx.doi.org/10.1371/journal.ppat.1009262
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