, ; and 100% specificity for AMI (85.4-100), , and compared to the BACTEC system. Comparing the results obtained in LJ we observed 100% sensitivity for RMP , followed by , , and 100% specificity for all drugs tested except for

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In this article, the first isolation of Mycobacterium kyorinense specimens in Brazil is described. M. kyorinense is a recently identified species, with a few strains reported only in Japan. The Brazilian isolates were initially identified as Mycobacterium celatum by PCR restriction enzyme pattern analysis (PRA) with hsp65. However, biochemical tests indicated the same profile of M. kyorinense and distinguished them from M. celatum and Mycobacterium branderi. The sequencing of the hsp65, rpoB, and 16S rRNA genes allowed the accurate identification of isolates as M. kyorinense. Mycobacterium kyorinense is a nonpigmented slowly growing mycobacterium that was first isolated from sputum of a patient with pneumonia. The species was described in 2009, and to date a few strains have been isolated, all from patients in Japan (5, 9). All patients were immunocompetent, and most of them had a history of pulmonary disease. M. kyorinense was considered a possible cause of clinically significant respiratory disease. The sequences of 16S rRNA genes, hsp65, and rpoB were identical in the strains tested but different from those of the two phylogenetically most related species, Mycobacterium celatum and Mycobacterium branderi (1, 4). Biochemical tests can also distinguish M. kyorinense from M. celatum and M. branderi (1,4,5,8).The aim of this work is to report the first isolation of M. kyorinense in Brazil and, to our knowledge, its first isolation outside Japan and to characterize its hsp65 restriction profile by PCR restriction enzyme pattern analysis (PRA).This study includes two pulmonary specimens (HF1629 and HF1836) that were initially identified as M. celatum in 2007 by PRA with hsp65 in the National Reference Laboratory for Tuberculosis of the Centro de Referência Professor Hélio Fraga/ENSP/ FIOCRUZ. These isolates were obtained from two separate expectorated sputum samples from a patient from Rio de Janeiro State, Brazil, according to microbiological diagnostic criteria for nontuberculous mycobacterial lung disease according to the American Thoracic Society guidelines (3).At admission to the health authority, the patient presented with complete destruction of the left lung by fibrotic lesions, which over the course of the disease extended to the right lung. Treatment with rifampin, isoniazid, pyrazinamide, and ethambutol temporarily improved the patient=s condition, but after less than 1 year, symptoms reappeared, leading to death.The DNA extraction protocol used in this work was developed in our laboratory and is being introduced here. Briefly, one loopful of bacteria grown on Lowenstein-Jensen medium was mixed in 300 l of 1% Triton X-100 and 100 l of acidwashed glass beads with a diameter of 106 m. The cells were shaken vigorously in a Vortex mixer for 1 min and incubated at room temperature for 8 min. After this incubation, 50 l of mixture was placed in a microtube with 100 l of lysis buffer (equal amounts of 15% Chelex 100, 0.5% Tween 20, and Tris-EDTA). The mixture was heated to 100°C for 20 min and shaken vigorously...

Three isolates of a slow-growing, non-chromogenic mycobacterium were grown from three sputum samples of a patient from the north-eastern Ceará state in Brazil. Identification at species level could not be obtained with PCR restriction analysis of the hsp65 gene. In order to characterize the isolates we carried out phenotypic and genotypic tests. We sequenced the nearly complete 16S rRNA gene and obtained partial sequences of the hsp65 (encoding the hypervariable region of the 65 kDa heat-shock protein) and rpoB (encoding the beta-subunit of RNA polymerase) genes. The three isolates turned out to be identical and most closely related to the species Mycobacterium celatum and Mycobacterium kyorinense. The results, however, showed significant differences between these species and the isolates studied, which led us to consider them members of a novel species for which we propose the name Mycobacterium fragae. The type strain is HF8705 T (5Fiocruz-INCQS/CMRVS P4051 T 5DSM 45731 T ).

(1) Background: The Commercial Kit SIRE Nitratase ® PlastLabor, is a drug susceptibility test kit used to detect Mycobacterium tuberculosis resistance to first-line TB treatment drugs. The present study aimed at evaluating its performance in a multicenter study. (2) Methods: To determine its accuracy, the proportion methods in Lowenstein Jensen medium or the BACTEC TM MGIT TM 960 system was used as a gold standard. (3) Results: The study revealed that the respective accuracies of the kit with 190 M. tuberculosis clinical isolates, using the proportion methods in Lowenstein Jensen medium or BACTEC TM MGIT TM 960 system as a gold standard, were 93.9% and 94.6%, 96.9% and 94.6%, 98.0% and 97.8%, and 98.0% and 98.9%, for streptomycin, isoniazid, rifampicin, and ethambutol, respectively. (4) Conclusion: Thus, the kit can rapidly screen resistance to streptomycin, isoniazid, rifampicin, and ethambutol. Additionally, it does not require sophisticated equipment; hence, it can be easily used in the laboratories of low and middle income countries.