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.
Highlights
Instituto Adolfo Lutz, Centro de Imunologia, Laboratório de Diagnóstico Molecular de Infecções Bacterianas, São Paulo, SP, Brasil
The mpt64 real-time polymerase chain reaction (PCR) assay showed an efficiency of 94.69%
All of the 127 Mycobacterium tuberculosis complex (MTC) isolates used in the evaluation of this assay before its implementation in the TB diagnosis routine were positive for mpt64
Summary
Instituto Adolfo Lutz, Centro de Imunologia, Laboratório de Diagnóstico Molecular de Infecções Bacterianas, São Paulo, SP, Brasil. Brazil is one of the high burden countries for tuberculosis, and a rapid diagnosis is essential for effective control of the disease. An in-house real-time polymerase chain reaction (PCR) assay targeting the mpt 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-hsp identification and to real-time PCR. 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. Brazil is one of the high TB burden countries, with more than 67,000 cases diagnosed in 2014, of which 23% were from the state of São Paulo (SP) (MS 2015). Phenotypic identification requires at least 15 days, whereas molecular diagnostics can be performed within hours (Espy et al 2006, Pfyffer 2007)
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