ArticleTuberculosis (Edinburgh, Scotland)2011
Rapid DNA extraction for specific detection and quantitation of Mycobacterium tuberculosis DNA in sputum specimens using Taqman assays.
Article in Tuberculosis (Edinburgh, Scotland), 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 13 citations in OpenAlex.
- Detection of circulating Mycobacterium tuberculosis-specific DNA by droplet digital PCR for vaccine evaluation in challenged monkeys and TB diagnosis.Emerging microbes & infections · 2018Article
- Article
- Pilot study of a rapid and minimally instrumented sputum sample preparation method for molecular diagnosis of tuberculosis.Scientific reports · 2016Article
- Evaluation of four molecular methods for the diagnosis of tuberculosis in pulmonary and blood samples from immunocompromised patients.Memorias do Instituto Oswaldo Cruz · 2014Article
- PCR to detect Mycobacterium tuberculosis in respiratory tract samples: evaluation of clinical data.Epidemiology and infection · 2014Article
- A missed tuberculosis diagnosis resulting in hospital transmission.Infection control and hospital epidemiology · 2014Article
- Validation of Mycobacterium tuberculosis Rv1681 protein as a diagnostic marker of active pulmonary tuberculosis.Journal of clinical microbiology · 2013Article
- Nucleic acid testing for tuberculosis at the point-of-care in high-burden countries.Expert review of molecular diagnostics · 2012Review
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
Abstract
Rapid tuberculosis (TB) detection is critical for disease control, and further quantitation of Mycobacterium tuberculosis (Mtb) in sputum is valuable for epidemiological and clinical studies. We evaluated a simple, robust and cost-efficient in-house DNA extraction and downstream Taqman approach for detection and quantitation of Mtb genomes from sputum of newly-diagnosed TB patients and non-TB controls. DNA was extracted using guanidine isothiocyanate and silica-based spin columns in less than 2 h, stored frozen, and Taqman assays were used to detect Mtb with IS6110 and quantify it targeting RD1 and IS1081. The Taqmans had a sensitivity >95% in 108 culture-confirmed TB patients and specificity of 100% in 43 non-TB controls. Genome counts were correlated with the Mycobacterial Growth Indicator Tubes' (MGIT) time-to-detection values (1/TTD × 1000; rho = 0.66; p < 0.001) in 91 TB patients (33 excluded with MGIT contamination). This linear relationship was nearly identical between mycobacteria isolated from sputum and H37Rv Mtb grown in-vitro to its log phase. TB treatment between 3 and 7 days was associated with lower 1/TTD × 1000 values but not with genome counts. Together, our protocol provides rapid, specific, inexpensive and quantitative detection of Mtb DNA in fresh or stored sputa making it a robust tool for prompt TB diagnosis, and with potential use for clinical and epidemiologic studies.
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