Multicentre study of nitrate reductase assay for rapid detection of rifampicin-resistant M. tuberculosis.

SETTING: Four regional laboratories belonging to the Mycobacteria Reference Laboratory of São Paulo State, Brazil. OBJECTIVE: To evaluate the nitrate reductase assay (NRA) for rifampicin (RMP) susceptibility testing of Mycobacterium tuberculosis directly from clinical sputum samples of patients with pulmonary tuberculosis (TB). DESIGN: Performance of the NRA for detection of M.tuberculosis susceptibility to RMP was evaluated with 210 clinical sputum samples received by the participating laboratories during 2005 and 2006 and compared with the results of the direct proportion method. RESULTS: Susceptibility tests performed using the NRA and the direct proportion method showed 204 susceptible isolates and six isolates resistant to RMP by both methods. NRA sensitivity and specificity for RMP was 100%. The NRA results of susceptibility tests against RMP were available in 15 days for 87% of the samples. The results showed that NRA may yield a rapid answer in determining resistance for the majority of sputum samples with smear results reported as 3+ and 2+. CONCLUSION: The results demonstrate the feasibility of NRA for screening resistant strains in sputum samples from patients with pulmonary TB. NRA represents a rapid and low-cost alternative method that might be used in microbiological laboratories where resources are scarce.

Contribution of the Ogawa-Kudoh swab culture method to the diagnosis of pulmonary tuberculosis in Brazil.

OBJECTIVE: To analyse the contribution of the Ogawa-Kudoh (O-K) swab culture method to the diagnosis of pulmonary tuberculosis (PTB) in four different regions of Brazil. DESIGN: This study was carried out in two phases. Phase 1 was designed to compare the direct swab culture method (O-K) with the culture concentrated method (N-acetyl-L-cysteine-sodium hydroxide [NALC-NaOH]); for this purpose, 569 sputum samples were cultured by both methods. Phase 2 was carried out to assess the contribution of the O-K method to the diagnosis of PTB in four different regions in Brazil, based on the evaluation of 19,163 sputum samples. RESULTS: In the first phase of the study, O-K culture had a sensitivity of 94.8% and specificity of 99.8% in cases confirmed by NALC-NaOH/Löwenstein-Jensen (LJ) culture. In the second phase of the study, the overall contribution of O-K culture compared to acid-fast bacilli (AFB) examination (AFB-/culture+) to the diagnosis of PTB was 29.8%. CONCLUSION: O-K culture contributes significantly to the diagnosis of smear-negative PTB. Importantly, this method allows the recovery of clinical isolates in areas where use of the standard culture centrifuge is impossible, indicating that the O-K swab culture method should become a standard method for TB diagnosis in these regions.