ArticlePloS one2020
Detection of drug resistant Mycobacterium tuberculosis by high-throughput sequencing of DNA isolated from acid fast bacilli smears.
Article in PloS one, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers, 1 of them a synthesis that pooled it.
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Who cites it
11 citing papers in PubMed, 1 synthesis or guideline pooled it, 13 citations in OpenAlex.
- Targeted next-generation sequencing to diagnose drug-resistant tuberculosis: a systematic review and meta-analysis.The Lancet. Infectious diseases · 2024Pooled it
- Implementation of sequencing-based surveillance for drug-resistant tuberculosis in Nigeria.Frontiers in tuberculosis · 2026Article
- Enhanced diagnosis of multi-drug-resistant microbes using group association modeling and machine learning.Nature communications · 2025Article
- Deep Amplicon Sequencing Reveals Culture-dependent Clonal Selection of Mycobacterium tuberculosis in Clinical Samples.Genomics, proteomics & bioinformatics · 2025Article
- Exploring gene mutations and multidrug resistance in Mycobacterium tuberculosis: a study from the Lung Hospital in Vietnam.Molecular biology reports · 2024Article
- Computational Approaches for the Structure-Based Identification of Novel Inhibitors Targeting Nucleoid-Associated Proteins in Mycobacterium Tuberculosis.Molecular biotechnology · 2024Article
- Pyrazinamide-resistant Tuberculosis Obscured From Common Targeted Molecular Diagnostics.Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy · 2023Article
- High Throughput Sequencing for Clinical Tuberculosis: An Overview.Pathogens (Basel, Switzerland) · 2022Review
- Experimental Confirmation that an UncommonAntimicrobial agents and chemotherapy · 2022Article
- Direct Molecular Detection of Drug-Resistant Tuberculosis from Transported Bio-Safe Dried Sputum on Filter-Paper.Current microbiology · 2022Article
- Revisiting the methods for detectingAccess microbiology · 2021Review
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Authors and funding
20 authors at 9 institutions in 6 countries.
Funding
Abstract
backgroundDrug 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.
methodsWe 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.
resultsWe 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.
conclusionThis rapid AFB smear DNA-based method for determining drug resistance may be useful for the diagnosis and surveillance of drug-resistant tuberculosis.
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