You are here

Int J Mycobacteriol DOI:10.1016/j.ijmyco.2015.12.001

Populations of latent Mycobacterium tuberculosis lack a cell wall: Isolation, visualization, and whole-genome characterization.

Publication TypeJournal Article
Year of Publication2016
AuthorsVelayati, AAkbar, Abeel, T, Shea, T, Zhavnerko, GKonstantin, Birren, B, Cassell, GH, Earl, AM, Hoffner, S, Farnia, P
JournalInt J Mycobacteriol
Volume5
Issue1
Pages66-73
Date Published2016 Mar
ISSN2212-554X
KeywordsCell Wall, Genome, Bacterial, High-Throughput Nucleotide Sequencing, Humans, Latent Tuberculosis, Mycobacterium tuberculosis
Abstract

OBJECTIVE/BACKGROUND: Mycobacterium tuberculosis (MTB) causes active tuberculosis (TB) in only a small percentage of infected people. In most cases, the infection is clinically latent, where bacilli can persist in human hosts for years without causing disease. Surprisingly, the biology of such persister cells is largely unknown. This study describes the isolation, identification, and whole-genome sequencing (WGS) of latent TB bacilli after 782days (26months) of latency (the ability of MTB bacilli to lie persistent).

METHODS: The in vitro double-stress model of latency (oxygen and nutrition) was designed for MTB culture. After 26months of latency, MTB cells that persisted were isolated and investigated under light and atomic force microscopy. Spoligotyping and WGS were performed to verify the identity of the strain.

RESULTS: We established a culture medium in which MTB bacilli arrest their growth, reduce their size (0.3-0.1μm), lose their acid fastness (85-90%) and change their shape. Spoligopatterns of latent cells were identical to original H37Rv, with differences observed at spacers two and 14. WGS revealed only a few genetic changes relative to the already published H37Rv reference genome. Among these was a large 2064-bp insertion (RvD6), which was originally detected in both H37Ra and CDC1551, but not H37Rv.

CONCLUSION: Here, we show cell-wall free cells of MTB bacilli in their latent state, and the biological adaptation of these cells was more phenotypic in nature than genomic. These cell-wall free cells represent a good model for understanding the nature of TB latency.

URLhttp://linkinghub.elsevier.com/retrieve/pii/S2212-5531(15)00164-8
DOI10.1016/j.ijmyco.2015.12.001
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/26927992?dopt=Abstract

Alternate JournalInt J Mycobacteriol
PubMed ID26927992
Grant ListU19 AI110818 / AI / NIAID NIH HHS / United States
HHSN272200900018C / / PHS HHS / United States
U19AI110818 / AI / NIAID NIH HHS / United States