Journal
AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE
Volume 91, Issue 4, Pages 709-714Publisher
AMER SOC TROP MED & HYGIENE
DOI: 10.4269/ajtmh.13-0603
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Funding
- Belgian Directorate-General for Development Cooperation (DGDC)
- Wellcome Trust
- charity IFHAD: Innovation For Health And Development
- Imperial College Biomedical Research Centre
- Joint Global Health Trials consortium (MRC, DFID, and Wellcome Trust)
- Medical Research Council [MR/K007467/1] Funding Source: researchfish
- MRC [MR/K007467/1] Funding Source: UKRI
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Real-time polymerase chain reaction (qPCR) was optimized for detecting Mycobacterium tuberculosis in sputum. Sputum was collected from patients (N = 112) with suspected pulmonary tuberculosis, tested by smear microscopy, decontaminated, and split into equal aliquots that were cultured in Lowenstein-Jensen medium and tested by qPCR for the small mobile genetic element 1S6110. The human ERV3 sequence was used as an internal control. 3 of 112 (3%) qPCR failed. For the remaining 109 samples, qPCR diagnosed tuberculosis in 79 of 84 patients with culture-proven tuberculosis, and sensitivity was greater than microscopy (94% versus 76%, respectively, P < 0.05). The qPCR sensitivity was similar (P = 0.9) for smear-positive (94%, 60 of 64) and smear-negative (95%, 19 of 20) samples. The qPCR was negative for 24 of 25 of the sputa with negative microscopy and culture (diagnostic specificity 96%). The qPCR had 99.5% sensitivity and specificity for 211 quality control samples including 84 non-tuberculosis mycobacteria. The qPCR cost similar to 5US$ per sample and provided same-day results compared with 2-6 weeks for culture.
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