Universidad Peruana Cayetano Heredia

Standardization of a TaqMan-based real-time PCR for the detection of Mycobacterium tuberculosis-complex in human sputum

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dc.contributor.author Barletta, Francesca
dc.contributor.author Vandelannoote, Koen
dc.contributor.author Collantes, Jimena
dc.contributor.author Evans, Carlton Anthony William
dc.contributor.author Arévalo Zelada, Jorge Luis
dc.contributor.author Rigouts, Leen
dc.date.accessioned 2020-06-10T18:11:35Z
dc.date.available 2020-06-10T18:11:35Z
dc.date.issued 2014
dc.identifier.uri https://hdl.handle.net/20.500.12866/8014
dc.description.abstract 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 IS6110. 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 approximately 5US$ per sample and provided same-day results compared with 2-6 weeks for culture. en_US
dc.language.iso eng
dc.publisher American Society of Tropical Medicine and Hygiene
dc.relation.ispartofseries American Journal of Tropical Medicine and Hygiene (ASTMH Journal)
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject DNA Primers/genetics en_US
dc.subject DNA, Bacterial/genetics en_US
dc.subject Humans en_US
dc.subject Mycobacterium tuberculosis/genetics/isolation & purification en_US
dc.subject Quality Control en_US
dc.subject Real-Time Polymerase Chain Reaction/economics/methods/standards en_US
dc.subject Reference Standards en_US
dc.subject Sensitivity and Specificity en_US
dc.subject Sputum/microbiology en_US
dc.subject Tuberculosis, Pulmonary/diagnosis/microbiology en_US
dc.title Standardization of a TaqMan-based real-time PCR for the detection of Mycobacterium tuberculosis-complex in human sputum en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.4269/ajtmh.13-0603
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#3.03.06
dc.relation.issn 1476-1645


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