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Serial image analysis of Mycobacterium tuberculosis colony growth reveals a persistent subpopulation in sputum during treatment of pulmonary TB

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Sloan_2016_SerialImageAnalysis_Tuberculosis_CC.pdf (661.9Kb)
Date
05/2016
Author
Barr, David A.
Kamdolozi, Mercy
Nishihara, Yo
Ndhlovu, Victor
Khonga, Margaret
Davies, Geraint R.
Sloan, Derek James
Keywords
Pharmacodynamics
Biomarkers
Persisters
Drug tolerance
RA0421 Public health. Hygiene. Preventive Medicine
NDAS
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Abstract
Faster elimination of drug tolerant ‘persister’ bacteria may shorten treatment of tuberculosis (TB) but no method exists to quantify persisters in clinical samples. We used automated image analysis to assess whether studying growth characteristics of individual Mycobacterium tuberculosis colonies from sputum on solid media during early TB treatment facilitates ‘persister’ phenotyping. As Time to Detection (TTD) in liquid culture inversely correlates with total bacterial load we also evaluated the relationship between individual colony growth parameters and TTD. Sputum from TB patients in Malawi was prepared for solid and liquid culture after 0, 2 and 4 weeks of treatment. Serial photography of agar plates was used to measure time to appearance (lag time) and radial growth rate for each colony. Mixed-effects modelling was used to analyse changing growth characteristics from serial samples. 20 patients had colony measurements recorded at ≥1 time-point. Overall lag time increased by 6.5 days between baseline and two weeks (p = 0.0001). Total colony count/ml showed typical biphasic elimination, but long lag time colonies (>20days) had slower, monophasic decline. TTD was associated with minimum lag time (time to appearance of first colony1). Slower elimination of long lag time colonies suggests that these may represent a persister subpopulation of bacilli.
Citation
Barr , D A , Kamdolozi , M , Nishihara , Y , Ndhlovu , V , Khonga , M , Davies , G R & Sloan , D J 2016 , ' Serial image analysis of Mycobacterium tuberculosis colony growth reveals a persistent subpopulation in sputum during treatment of pulmonary TB ' , Tuberculosis , vol. 98 , pp. 110-115 . https://doi.org/10.1016/j.tube.2016.03.001
Publication
Tuberculosis
Status
Peer reviewed
DOI
https://doi.org/10.1016/j.tube.2016.03.001
ISSN
1472-9792
Type
Journal article
Rights
© 2016 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Description
This study received no direct funding. D.A.B. is supported by a Wellcome Trust Clinical PhD studentship (105165/Z/14/A). Tuberculosis Laboratory, College of Medicine, Malawi is supported by PanACEA Biomarker Extension Programe (PANBIOME) (SP.2011.41304.008).
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  • University of St Andrews Research
URI
http://hdl.handle.net/10023/8668

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