ABSTRACT
We evaluated a multiplex-PCR to differentiate Mycobacterium bovis from M. tuberculosis Complex (MTC) by one step amplification based on simultaneous detection of pncA 169 C > G change in M. bovis and the IS6110 present in MTC species. Our findings showed the proposed multiplex-PCR is a very useful tool for complementation in differentiating M. bovis from other cultured MTC species.
Subject(s)
Bacteriological Techniques/methods , Molecular Diagnostic Techniques/methods , Multiplex Polymerase Chain Reaction/methods , Mycobacterium bovis/isolation & purification , Mycobacterium tuberculosis/isolation & purification , Tuberculosis/microbiology , Amidohydrolases/genetics , DNA Transposable Elements , DNA, Bacterial/genetics , Mycobacterium bovis/classification , Mycobacterium bovis/genetics , Mycobacterium tuberculosis/classification , Mycobacterium tuberculosis/genetics , Tuberculosis/diagnosisABSTRACT
We evaluated a multiplex-PCR to differentiate Mycobacterium bovis from M. tuberculosis Complex (MTC) by one step amplification based on simultaneous detection of pncA 169C > G change in M. bovis and the IS6110 present in MTC species. Our findings showed the proposed multiplex-PCR is a very useful tool for complementation in differentiating M. bovis from other cultured MTC species.
Subject(s)
Bacteriological Techniques/methods , Molecular Diagnostic Techniques/methods , Multiplex Polymerase Chain Reaction/methods , Mycobacterium bovis/isolation & purification , Mycobacterium tuberculosis/isolation & purification , Tuberculosis/microbiology , Amidohydrolases/genetics , DNA Transposable Elements , DNA, Bacterial/genetics , Mycobacterium bovis/classification , Mycobacterium bovis/genetics , Mycobacterium tuberculosis/classification , Mycobacterium tuberculosis/genetics , Tuberculosis/diagnosisABSTRACT
The microscopic observation drug susceptibility assay (MODS) was evaluated to determine susceptibility to pyrazinamide in Mycobacterium tuberculosis, and compared with the broth microdilution method (BMM), absolute concentration method (ACM), and pyrazinamidase (PZase) determination. We tested 34 M. tuberculosis clinical isolates (24 sensitive and eight resistant to pyrazinamide) and the control strains M. tuberculosis H37Rv (ATCC 27294) and Mycobacterium bovis AN5. The MODS, BMM, ACM and PZase determination provided results in average times of 6, 18, 28 and 7 days, respectively. All methods showed excellent sensitivity and specificity (p <0.05). Of the methods studied, the MODS proved to be faster, efficient, inexpensive, and easy to perform. However, additional studies evaluating the MODS in differentiating pyrazinamide-resistant and pyrazinamide-susceptible M. tuberculosis must be conducted with a larger number of clinical isolates.
