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Title
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Standardization of a TaqMan-based real-time PCR for the detection of mycobacterium tuberculosis-complex in human sputum
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Author
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Abstract
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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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Language
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English
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Source (journal)
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The American journal of tropical medicine and hygiene. - Baltimore, Md
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Publication
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Baltimore, Md
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2014
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ISSN
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0002-9637
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DOI
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10.4269/AJTMH.13-0603
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Volume/pages
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91
:4
(2014)
, p. 709-714
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ISI
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000342957600011
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Pubmed ID
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25114009
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Full text (Publisher's DOI)
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