Alternative sputum preparation to improve polymerase chain reaction assay for Mycobacterium tuberculosis detection
OBJECTIVE: To improve polymerase chain reaction (PCR) sensitivity in detecting Mycobacterium tuberculosis in sputum samples.
METHODS: This study proposed the use of glass beads as a modification of the routine protocol for sputum preparation used in the Laboratory of Molecular Biology and Mycobacteria at the Federal University of Santa Catarina, Florianópolis, Brazil. The study comprised 120 sputum samples, 60 of which were treated with the routine protocol, while 60 were treated with the modified protocol using glass beads.
RESULTS: Samples treated with the routine protocol had a sensitivity of 56.7% (95%CI 44.1–69.2) in 16S rRNA PCR and 81.7% (95%CI 71.9–91.5) in insertion sequence (IS) 6110 PCR, compared with culture. Samples treated with the modified protocol had a sensitivity of 73.3% (95%CI 62.1–84.5) in 16S rRNA PCR and 100% in IS6110 PCR.
CONCLUSION: The modified protocol using glass beads greatly improved mycobacterial detection in sputum samples compared with the routine protocol.
Document Type: Research Article
Affiliations: 1: Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil 2: Laboratório Central do Estado de Santa Catarina, Florianópolis, Santa Catarina, Brazil 3: Hospital Nereu Ramos, Florianópolis, Santa Catarina, Brazil 4: Universidade Federal de Santa Catarina, Florianópolis, Santa Catarina, Brazil; Universidade do Sul de Santa Catarina, Santa Catarina, Brazil
Publication date: 2012-06-01
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