Imperial College London
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Professor Carlton A W Evans

Faculty of Medicine, Department of Infectious Disease

Professor of Global Health
 
 
 
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Contact

 

+44 (0)20 3313 3222carlton.evans Website

 
 
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Location

 

Commonwealth BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Rowneki:2020:10.1371/journal.pone.0232343,
author = {Rowneki, M and Aronson, N and Du, P and Sachs, P and Blakemore, R and Chakravorty, S and Levy, S and Jones, AL and Trivedi, G and Chebore, S and Addo, D and Byarugaba, DK and Njobvu, PD and Wabwire-Mangen, F and Erima, B and Ramos, ES and Evans, CA and Hale, B and Mancuso, JD and Alland, D},
doi = {10.1371/journal.pone.0232343},
journal = {PLoS One},
title = {Detection of drug resistant Mycobacterium tuberculosis by high-throughput sequencing of DNA isolated from acid fast bacilli smears},
url = {http://dx.doi.org/10.1371/journal.pone.0232343},
volume = {15},
year = {2020}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - BACKGROUND: Drug susceptibility testing for Mycobacterium tuberculosis (MTB) is difficult to perform in resource-limited settings where Acid Fast Bacilli (AFB) smears are commonly used for disease diagnosis and monitoring. We developed a simple method for extraction of MTB DNA from AFB smears for sequencing-based detection of mutations associated with resistance to all first and several second-line anti-tuberculosis drugs. METHODS: We isolated MTB DNA by boiling smear content in a Chelex solution, followed by column purification. We sequenced PCR-amplified segments of the rpoB, katG, embB, gyrA, gyrB, rpsL, and rrs genes, the inhA, eis, and pncA promoters and the entire pncA gene. RESULTS: We tested our assay on 1,208 clinically obtained AFB smears from Ghana (n = 379), Kenya (n = 517), Uganda (n = 262), and Zambia (n = 50). Coverage depth varied by target and slide smear grade, ranging from 300X to 12000X on average. Coverage of ≥20X was obtained for all targets in 870 (72%) slides overall. Mono-resistance (5.9%), multi-drug resistance (1.8%), and poly-resistance (2.4%) mutation profiles were detected in 10% of slides overall, and in over 32% of retreatment and follow-up cases. CONCLUSION: This rapid AFB smear DNA-based method for determining drug resistance may be useful for the diagnosis and surveillance of drug-resistant tuberculosis.
AU  - Rowneki,M
AU  - Aronson,N
AU  - Du,P
AU  - Sachs,P
AU  - Blakemore,R
AU  - Chakravorty,S
AU  - Levy,S
AU  - Jones,AL
AU  - Trivedi,G
AU  - Chebore,S
AU  - Addo,D
AU  - Byarugaba,DK
AU  - Njobvu,PD
AU  - Wabwire-Mangen,F
AU  - Erima,B
AU  - Ramos,ES
AU  - Evans,CA
AU  - Hale,B
AU  - Mancuso,JD
AU  - Alland,D
DO  - 10.1371/journal.pone.0232343
PY  - 2020///
SN  - 1932-6203
TI  - Detection of drug resistant Mycobacterium tuberculosis by high-throughput sequencing of DNA isolated from acid fast bacilli smears
T2  - PLoS One
UR  - http://dx.doi.org/10.1371/journal.pone.0232343
UR  - https://www.ncbi.nlm.nih.gov/pubmed/32384098
UR  - http://hdl.handle.net/10044/1/79482
VL  - 15
ER  -
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