Journal
SCIENTIFIC REPORTS
Volume 7, Issue -, Pages -Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/s41598-017-10234-z
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Funding
- PreDiCT-TB consortium [IMI Joint undertaking grant from the European Union's Seventh Framework Programme (FP7)] [115337]
- PreDiCT-TB consortium [EFPIA companies']
- UK Engineering and Physical Sciences Research Council [EP/J01771X/1]
- European Union FAMOS project (FP7 ICT) [317744]
- Royal Society Leverhulme Trust Senior Fellowship
- Department of Health, UK
- Engineering and Physical Sciences Research Council [EP/J01771X/1] Funding Source: researchfish
- EPSRC [EP/J01771X/1] Funding Source: UKRI
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Tuberculosis relapse is a barrier to shorter treatment. It is thought that lipid rich cells, phenotypically resistant to antibiotics, may play a major role. Most studies investigating relapse use sputum samples although tissue bacteria may play an important role. We developed a non-destructive, labelfree technique combining wavelength modulated Raman (WMR) spectroscopy and fluorescence detection (Nile Red staining) to interrogate Mycobacterium tuberculosis cell state. This approach could differentiate single dormant (lipid rich, LR) and non-dormant (lipid poor, LP) cells with high sensitivity and specificity. We applied this to experimentally infected guinea pig lung sections and were able to distinguish both cell types showing that the LR phenotype dominates in infected tissue. Both in-vitro and ex-vivo spectra correlated well, showing for the first time that Mycobacterium tuberculosis, likely to be phenotypically resistant to antibiotics, are present in large numbers in tissue. This is an important step in understanding the pathology of relapse supporting the idea that they may be caused by M. tuberculosis cells with lipid inclusions.
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