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Capture and visualization of live Mycobacterium tuberculosis bacilli from tuberculosis bioaerosols

By Ryan Dinkele, Sophia Gessner, Anastasia S. Koch, Carl Morrow, Melitta Gqada, Mireille Kamariza, Carolyn R Bertozzi, Brian Smith, Courtney McLoud, Andrew Kamholz, Wayne Bryden, Charles Call, Valerie Mizrahi, Robin Wood, Digby Francis Warner

Posted 27 Dec 2019
bioRxiv DOI: 10.1101/2019.12.23.887729

The tuberculosis (TB) pandemic demands urgent interventions such as those designed to interrupt Mycobacterium tuberculosis ( Mtb ) transmission, a challenge exacerbated by our poor understanding of the events enabling successful transfer of infectious bacilli between hosts. To address this problem, we developed the Respiratory Aerosol Sampling Chamber (RASC), a personal clean-room equipped with high-efficiency filtration and sampling technologies that allow biosafe capture and isolation of particulate matter, including Mtb bacilli, released by patients during natural breathing and (non-induced) cough. Here, we demonstrate the use of DMN-trehalose labelling to detect and quantify live Mtb bacilli among complex bioaerosol samples arrayed in a bespoke nanowell device following capture in the RASC. A pilot study identified Mtb in more than 85 % of known TB-patients, improving significantly on previous work which has relied on animal infection and cough sampling to estimate transmission events. Moreover, intra-patient comparisons of bioaerosol and sputum samples indicated that Mtb aerosols likely derive from a compartment other than sputum. These results support the utility of the RASC platform for research aimed at interrupting Mtb transmission, including the non-invasive detection of Mtb -infected individuals who are predicted to contribute to bacillary spread despite the absence of clinical symptoms.

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