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

Collantes, Jimena; Barletta, Francesca; Rigouts, Leen; Arévalo Zelada, Jorge Luis; Evans, Carlton Anthony William; Vandelannoote, Koen

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.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