Clinical isolates of M. tuberculosis (n 330) were studied using MIRUVNTR methods, based on the analysis of 35 loci and analysis of point mutations. It is shown that each branch of the dendrogram of the molecular genetic relatedness of M. tuberculosis clinical isolates
contains isolates relating to the only one cluster group, defined by the point mutations analysis. All isolates of M. tuberculosis, relating to Beijing spoligofamily are in the same branch of the dendrogram. Isolates from the other spoligofamilies are missing in the group.
Isolates, belonging to LAM spoligofamily, are in the same branch of the dendrogram, which also includes isolates, belonging to T and S spoligofamilies. Isolates from T and Haarlem spoligo families comprise into several branches of the dendrogram, and T spoligofamily isolates belong to three SNP groups, while Haarlem isolates belong to the only one group, identified on the basis of point mutations analysis.

According to suggested scheme genotyping of M. tuberculosis in Ural region of Russia is based on 2-step assay: the real-time PCR for differentiation of Beijing/non-Beijing isolates and further discrimination by MIRU-VNTR-typing using a set of 9 loci (MIRU26, QUB26, Mtub21, Qub11b, MIRU31, MIRU40 VNTR4120, VNTR3820, VNTR3232) for Beijing isolates, and 15 loci (Mtub04, ETRC, MIRU04, MIRU40, MIRU10, MIRU16, Mtub21, QUB11b, ETRA, Mtub30, MIRU26, MIRU31, Mtub39, QUB26, QUB4156) and/or spoligotyping for non-Beijing isolates.

The preliminary results of the use of semi-quantitative immunoassay method (BRAHMSPCT-Q) for rapid diagnosis of procalcitonin level in 35 children and adolescents with severe tuberculosis are presented. Positive result of the procalcitonin test (more than 0,5 ng/ml) has been obtained in 12 patients, and negative one (less than 0,5 ng / ml) – in 23. Positive test result indicates high activity of mycobacterial population, confirmed by the detection of mycobacteria using three methods in such patients: fluorescent microscopy, cultural method on BACTEC and PCR diagnostics(83,3%, 100% and 83,3%, respectively) in contrast to the group of patients with negative test result (17,4%, 34,8% and 47,8%, respectively). In the patients with positive procalcitonin test, reliably greater production of some proinflammatory cytokines, MIF, TNF-α and IFN-γ, has been revealed.

The features of the tissue and cellular responses of the respiratory system at different severity of systemic inflammatory response syndrome (SIRS) were studied in 36 operated patients with destructive pulmonary MDR TB. It was found that extensive specific lesion of the lung tissue with predominance of exudative alterative reaction and development of extensive seroplastic inflammation in parenchyma areas, free from caseation, took place in a pronounced SIRS. The destruction of air-blood barrier components, dysfunction of pulmonary surfactant production by the type 2 alveolocytes, metaplasia of flat alveolar epithelium in the cube one, appearance of a significant number of lipophages and erythrophages, eosinophilic material and comprising strands of fibrin in the intra-alveolar content could be observed. The findings suggest that the correlation of the main SIRS indicators with structural changes of microcirculation, the tissue and cellular responses of the respiratory system exists, and indicate the possibility of the diffusive alveolar damage development in patients with acute progressing of destructive pulmonary tuberculosis caused by MDR MBT.

104 sputum samples obtained from the patients with pulmonary tuberculosis in the Central Research Institute for Tuberculosis of the Russian Academy of Medical Sciences were studied in order to compare the results of MBT detection and identification of drug susceptibility to rifampicin using test system Xpert MBT/RIF and classical microbiological methods. In addition, the aim of the research was to study the effect of concentrating of diagnostic specimens to an increase of the test Xpert MBT/RIF sensitivity. The Xpert MBT/RIF demonstrated most sensitivity with positive results in 78,2% of cases to compare with the native light microscopy of sputum, detecting 48,5% of cases, fluorescent microscopy of sputum sediment – 67,3%, cultural method with Lowenstein-Jensen medium – 56,4% and on BACTEC MGIT 960 – 71,3%.Concentration of sputum samples had no significant effect on the increase of the test sensitivity. Samples with identified non-tuberculous mycobacteria by the test Xpert MTB/RIF were recorded as negative ones, showing high specificity of the method. Complete coincidence of the results of rifampicin resistance detection using biological microchips and the system Xpert MBT/RIF was found.

Timely administration of appropriate treatment reduces the level and rate of emergence of resistant strains of mycobacteria. Therefore, the actual is to develop methods for early and reliable drug susceptibility testing. With the help of test systems Sensititre MycoTB
studied 149 strains of M. tuberculosis, isolated from respiratory material patients with chronic forms of tuberculosis. With the help of test systems Sensititre SLOMyco RAPMyco were tested 471 NTMB strain isolated from patients with suspected tuberculosis or mycobacteriosis belonging to species: MAC – 139, M. kansasii – 57, M. xenopi – 59, M. fortuitum group – 170 and M. chelonae group – 46.
The results allowed to conclude that the test system Sensititre have advantages associated with the standardization process and to determine the extent susceptibility/resistant isolates of mycobacteria, as well as the possibility of obtaining information about the spectrum of drug sensitivity to a large number of drugs in the short term.

The article presents original data on strain sensitivity determination of C. albicans, C dubliniensis, C. glabrata, C. guilliermondii, C. kefyr, C. krusei, C. lipolytica, C. lusitaniae, C. parapsilosis, C. rugosa, C. tropicalisand C. zeylanoides to antifungal drugs. Activity of amphotericin B, fluconazole, itraconazole, voriconazole, flucytosine (5-fluorocytosine) against Candidastrains, isolated in the tuberculosis clinic, was analyzed.

The results of local microbiological monitoring at the secondary infections, caused by opportunistic micro flora in patients with pulmonary tuberculosis, carried out 2010-2012 in Moscow are presented. Gram-positive cocci Streptococcus gr. viridian have been identified as leading microorganisms. Comparative analysis of the microorganisms resistance dynamics in vitro to broadspectrum antibiotics with anti-TB activity – rifampicin, levofloxacin, moxifloxacin, was performed. It demonstrated the difference in the profile of resistance of S. gr. viridian strains, isolated from the patients, examined in the branches and subdivisions of the Moscow Research and Clinical Center for Tuberculosis Control of the Moscow Government Health Department.

Aim: to compare results of skin testing using the preparation CFP-10-ESAT-6 (DST) and QuantiFERON – GIT (QFT) in children and adolescents with local TB or latent TB infection (LTBI). 210 children and adolescents were studied including 163 (77,6%) with TB and 47 (22,4%) with LTBI. All the children received chemotherapy for at least 2 months for LTBI and 6 months for active TB. Positive DST was observed in 172 (81,9%, 95%CI 76,1-86,6%) including 90,7% in TB-patients and 51,1% in LTBI, р<0,001. Positive QFT-GIT was registered in 176 (83,8%, 95%CI 78,2-88,2%) subjects including 92,6% in TB-patients and 53,2% in LTBI, р<0,001. Agreement of DST and QFT-GIT was 90,5% (95%CI 85,7-93,8%), kappa coefficient was 0,67±0,07 (р<0,001). Discordance was registered in 20 (9,5%) cases. 11 patients with negative DST reactions have positive QFT results. Among them 9 patients initially had positive DST – reactions and after chemotherapy reaction became negative. Probably DST reversion in these children as a result of chemotherapy pass ahead of reversion QFT results. This determined the discordant reactions.

A study to assess the stability of the diagnostic product DIASKINTEST®– TB recombinant allergen in the standard dilution, solution for intradermal administration, in terms of «specific activity» and «specificity» in guinea pigs, according to the requirements of FSP LSR-006435/08-110808, depending on storage temperature and time, was performed. The absence of significant differences in the specific activity and specificity of the DIASKINTEST®was founddepending on the storage temperature in the range of 2°C to 23°C and the duration of storage of 7 to 30 days.

Considering the growing outspread of drug-resistant strains of M. tuberculosis, a clear understanding of the molecular mechanisms underlying drug-resistance of the pathogen to different groups of antibiotics is crucial. Fluoroquinolone-resistant M. tuberculosis generally harbor various mutations in the gyrA and gyrB genes, mostly in the QRDR region. M. tuberculosis resistance to aminoglycosides/ capreomycin is commonly associated with mutations in the rrs, gene, the eis promoter region, and occasionally in the tlyA gene. Two assays are available for the detection of mutations leading to fluoroquinolone and aminoglycoside resistance in clinical practice: the TB-BIOCHIP2 (only for fluoroquinolones) and the GenoType MTBDRsl.