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Memórias do Instituto Oswaldo Cruz
Fundação Oswaldo Cruz, Fiocruz
ISSN: 1678-8060 EISSN: 1678-8060
Vol. 111, No. 9, 2016, pp. 545-550
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Bioline Code: oc16084
Full paper language: English
Document type: Research Article
Document available free of charge
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Memórias do Instituto Oswaldo Cruz, Vol. 111, No. 9, 2016, pp. 545-550
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Performance of an in-house real-time polymerase chain reaction for identification of Mycobacterium tuberculosis isolates in laboratory routine diagnosis from a high burden setting
Gallo, Juliana Failde; Pinhata, Juliana Maira Watanabe; Chimara, Erica; Gonçalves, Maria Gisele; Fukasawa, Lucila Okuyama & Oliveira, Rosangela Siqueira de
Abstract
Brazil is one of the high burden countries for tuberculosis, and a rapid diagnosis is essential for effective control
of the disease. In the present study, an in-house real-time polymerase chain reaction (PCR) assay targeting
the mpt64 gene for identification of Mycobacterium tuberculosis complex isolates was evaluated under routine
diagnosis conditions in a reference laboratory. From May 2011 to July 2012, 1,520 isolates of mycobacteria were
prospectively submitted for phenotypic and/or PRA-hsp65 identification and to real-time PCR. The mpt64 real-time
PCR showed 99.7% sensitivity and 96% specificity and detected 79.4% of the cases missed by phenotypic and PRA-hsp65
identification. The in-house real-time PCR assay showed high sensitivity and specificity and was successfully
implemented in the routine diagnosis of tuberculosis in a reference laboratory from a high burden setting.
Keywords
molecular diagnostics; MPT64 protein; Mycobacterium tuberculosis; real-time polymerase chain reaction; tuberculosis
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