ObjectiveThe objective of the research was to assess the susceptibility of the slowly growing nontuberculous mycobacteria strains to the antimicrobial drugs used for mycobaterioses treatment using SLOMYCO test system.Materials and methodsWe assessed 363 NTM strains: 177 MAC (161 M. avium, 16 M. intracellulare), 112 M. kansasii and 74 M. xenopi collected from the respiratory material of the patients were under the treatment or under diagnostic procedures at our Center, affiliates and the diagnostic department in 2010–2016. Drug sucseptibility for NTM was tested using the Sensititre SLOWMYCO system (TREK DIAGNOSTIC Systems Ltd., UK). MICs were established by microdilutions in Mueller-Hinton broth on polystyrene 96-well plates. The statistical analysis was done using the StatGraphics Plus 5.0 software. The data were compared pairwise using Pearson χ2 test with Yates correction. 95% confidence interval (CI) were calculated. Statistically significant differences were considered for p <0.05. Log-rank test and Kaplan-Meier curves were used to assess the concentration-dependent surveillance probability.ResultsThe statistically significant differences were revealed in sensitivity/resistance isolates of M. avium and M. intracellulare: M. avium strains were resistant to higher concentrations of amikacin, clarithromycin, linezolid and streptomycin (p <0.01); M. intracellulare strains were resistant to higher concentrations of ethionamide (p <0.05). The isolates of M. avium were significantly more resistant than M. kansasii to amikacin, doxycycline, isoniazid, clarithromycin, linezolid, moxifloxacin, rifabutin, rifampicin, streptomycin, trimethoprim/sulfamethoxazole, ciprofloxacin, ethambutol, ethionamide (visible growth of M. avium were inhibited by higher drug concentrations, p <0.01). The isolates of M. avium showed significantly higher resistance than M. xenopi to amikacin, doxycycline, isoniazid, clarithromycin, linezolid, moxifloxacin, rifampicin, streptomycin, trimethoprim/sulfamethoxazole, ciprofloxacin, ethambutol, and ethionamide (visible growth of M. avium were inhibited by higher drug concentrations, p <0.01). Statistically significant differences in the dynamics of the response to the antibacterial effects of isoniazid, linezolid, moxifloxacin, rifampicin, trimethoprim/sulfamethoxazole, ethambutol, and ethionamide were found for M. intracellulare and M. xenopi (complete inhibition of the visible growth of M. intracellulare required higher drugs concentrations, p <0, 05). Comparison of the Kaplan-Meyer curves revealed statistically significant differences in survialence probability of M. kansasii and M. xenopi for amikacin, doxycycline, rifampicin, trimethoprim/sulfamethoxazole, ciprofloxacin, ethambutol, and ethionamide (a higher number of isolates of M. xenopi were inhibited by low drugs concentrations, p <0.05).ConclusionsOur data show that M. avium and M. intracellulare were more resistant to the majority of the studied drugs than M. kansasii and M. xenopi.

Three-component seismograms of small local earthquakes recorded in the Peter the First Range of mountains near Garm, Tadzhikistan SSR, display shear-wave splitting similar to that previously observed near the North Anatolian Fault in Turkey. The Peter the First Range is in a region of compressional tectonics, whereas the North Anatolian Fault is a comparatively simple strike-slip fault. Detailed analysis of the Turkish records suggests that the splitting is diagnostic of crack-induced anisotropy caused by vertical microcracks aligned parallel to the direction of maximum compression. Preliminary examination of paper records from Garm shows that most shear waves arriving within the shear-wave window display shear-wave splitting, and that the polarizations of leading shear-waves are consistently aligned in a NE/SW direction. The area is complicated and the tectonics are not wellunderstood, but the NE/SW direction is approximately perpendicular t o the compressional axis in many of the fault-plane mechanisms of the earthquakes. These earthquakes are usually at depths between 5 and 12 km, although there are some deeper events nearby.Parallel shear-wave polarizations, such as those observed, are expected to indicate the strike of nearly vertical parallel microcracks, which would be aligned parallel to the direction of maximum compression. Thus the shearwave polarizations in the Peter the First Range indicate that the directions of principal stress are reversed in the rock above the earthquake foci where thrust faulting is taking place.

Federation eIn addition to the obligatory pathogenic species of the Mycobacterium tuberculosis complex and Mycobacterium leprae, the genus Mycobacterium also includes conditionally pathogenic species that in rare cases can lead to the development of nontuberculous mycobacterial diseases. Because tuberculosis and mycobacteriosis have similar clinical signs, the accurate identification of the causative agent in a clinical microbiology laboratory is important for diagnostic verification and appropriate treatment. This report describes a low-density hydrogel-based microarray containing oligonucleotide probes based on the species-specific sequences of the gyrB gene fragment for mycobacterial species identification. The procedure included the amplification of a 352-nucleotide fragment of the gene and its hybridization on a microarray. The triple-species-specific probe design and the algorithm for hybridization profile recognition based on the calculation of Pearson correlation coefficients, followed by the construction of a profile database, allowed for the reliable and accurate identification of mycobacterial species, including mixed-DNA samples. The assay was used to evaluate 543 clinical isolates from two regions of Russia, demonstrating its ability to detect 35 mycobacterial species, with 99.8% sensitivity and 100% specificity when using gyrB, 16S, and internal transcribed spacer (ITS) fragment sequencing as the standard. The testing of clinical samples showed that the sensitivity of the assay was 89% to 95% for smear-positive samples and 36% for smear-negative samples. The large number of identified species, the high level of sensitivity, the ability to detect mycobacteria in clinical samples, and the up-to-date profile database make the assay suitable for use in routine laboratory practice. In addition to the obligatory pathogenic species of the Mycobacterium tuberculosis complex (MTC) and Mycobacterium leprae, the genus Mycobacterium also includes conditionally pathogenic species that in rare cases lead to the development of nontuberculous mycobacterial disorders in humans (1). The main risk factors for such diseases are immunosuppression and chronic obstructive pulmonary disease (2). Because tuberculosis and mycobacteriosis have common clinical signs, the proper identification of the causative agent in a clinical microbiology laboratory is one of the most important tasks for diagnostic verification and treatment with the correct medication (3).The conventional microbiological and biochemical methods used to detect Mycobacterium are labor-intensive and timeconsuming. Efficient mycobacterial species identification using high-performance liquid chromatography of mycolic acids (4) requires expensive equipment and high-level personnel qualifications. Moreover, the spectrum of species that can be identified by microbiological assays is rather narrow, making DNA-based methods preferable (5). A number of molecular genetic techniques have been developed, including in situ hybridization species-specific probes (6), mult...

No abstract

BackgroundThe goal of this study was to compare the consistency of three assays for the determination of the drug resistance of Mycobacterium tuberculosis (MTB) strains with various resistance profiles isolated from the Moscow region.MethodsA total of 144 MTB clinical isolates with a strong bias toward drug resistance were examined using Bactec MGIT 960, Sensititre MycoTB, and a microarray-based molecular assay TB-TEST to detect substitutions in the rpoB, katG, inhA, ahpC, gyrA, gyrB, rrs, eis, and embB genes that are associated with resistance to rifampin, isoniazid, fluoroquinolones, second-line injectable drugs and ethambutol.ResultsThe average correlation for the identification of resistant and susceptible isolates using the three methods was approximately 94%. An association of mutations detected with variable resistance levels was shown. We propose a change in the breakpoint minimal inhibitory concentration for kanamycin to less than 5 μg/ml in the Sensititre MycoTB system. A pairwise comparison of the minimal inhibitory concentrations (MICs) of two different drugs revealed an increased correlation in the first-line drug group and a partial correlation in the second-line drug group, reflecting the history of the preferential simultaneous use of drugs from these groups. An increased correlation with the MICs was also observed for drugs sharing common resistance mechanisms.ConclusionsThe quantitative measures of phenotypic drug resistance produced by the Sensititre MycoTB and the timely detection of mutations using the TB-TEST assay provide guidance for clinicians for the choice of the appropriate drug regimen.