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.
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.
Instituto Adolfo Lutz, Centro de Bacteriologia, Núcleo de Tuberculose e Micobacterioses, São Paulo, SP, Brasil.
Juliana Failde
JF
Gallo
Instituto Adolfo Lutz, Centro de Bacteriologia, Núcleo de Tuberculose e Micobacterioses, São Paulo, SP, Brasil.
Juliana Maira Watanabe
JM
Pinhata
Instituto Adolfo Lutz, Centro de Bacteriologia, Núcleo de Tuberculose e Micobacterioses, São Paulo, SP, Brasil.
Erica
E
Chimara
Instituto Adolfo Lutz, Centro de Imunologia, Laboratório de Diagnóstico Molecular de Infecções Bacterianas, São Paulo, SP, Brasil.
Maria Gisele
MG
Gonçalves
Instituto Adolfo Lutz, Centro de Imunologia, Laboratório de Diagnóstico Molecular de Infecções Bacterianas, São Paulo, SP, Brasil.
Lucila Okuyama
LO
Fukasawa
Instituto Adolfo Lutz, Centro de Bacteriologia, Núcleo de Tuberculose e Micobacterioses, São Paulo, SP, Brasil.
Rosangela Siqueira de
RS
Oliveira
D000942:Antigens, Bacterial / Q000235:genetics*; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors; D014376:Tuberculosis / Q000175:diagnosis*
D000942:Antigens, Bacterial; C510537:MPT64 protein, Mycobacterium tuberculosis