[The detection of medication sensitivity of M.tuberculosis isolated from patients with tuberculosis with multiple medication resistance in testsystem "Sensititre MycoTB".]

The test-system "Sensititre MycoTB" was applied to analyze 159 cultures of M.tuberculosis isolated from patients with tuberculosis. According data of investigation in "Bactec 960" they are identified as cultures with multiple medicinal sensitivity. The detailed analysis of results of application of mentioned test-system (mycoTB) demonstrated that significant part of cultures with multiple medicinal sensitivity is sensitive to anti-tuberculosis pharmaceuticals. The significant part of cultures is characterized by "intermediate" sensitivity/resistance than can be established by mentioned test-system. This data permits enlarging possibilities of chemotherapy of tuberculosis.

[DETERMINATION OF CRITICAL CONCENTRATION OF CHEMOTHERAPY DRUGS FOR MYCOBACTERIUM TUBERCULOSIS SENSITIVITY EVALUATION USING Sensititre MyCoTB TEST SYSTEM].

AIM: Determination of critical concentration for chemotherapy drugs, widely used for tuberculosis treatment, for use in Mycobacterium tuberculosis drug sensitivity results evaluation by Sensititre MycoTB test-system. MATERIALS AND METHODS: Minimal inhibiting concentration (MIC) of isoniazid, rifampicin, streptomycin, ethambutol, amikacin, kanamycin, ofloxacin and moxifloxacin against conditionally sensitive and conditionally resistant strains of tuberculosis mycobacteria (TBM), isolated from various diagnostic material, obtained from patients with various forms of lung tuberculosis being treated in MCSPCTC hospital and dispensaries for tuberculosis control of Moscow, were studied in Sensititre MycoTB test system. RESULTS: Critical concentration of chemotherapy drugs for MycoTB test system was determined as a result of the obtained MIC values analysis as the minimal concentration that suppressed growth of 95% of sensitive strains and does not obstruct growth of 95% of resistant. The following MIC values were established: streptomycin--1.0, isoniazid--0.25, rifampicin--1.0, ethambutol--4.0, ofloxacin--2.0, moxifloxacin--0.25, kanamycin--2.5 and amikacin- 1.0 µg/ml. CONCLUSION: The developed critical concentration of the mentioned preparations is currently used for evaluation of sensitivity/ resistance of TBM clinical isolates from MCSPCTC.

[Determination of multidrug resistance of M. tuberculosis by different methods].

Testing the direct nitrate reductase technique versus the absolute concentration test has indicated that the former may be successfully used for rapid determination of the sensitivity of Mycobacterium tuberculosis (MBT) to isoniazid and rifampicin and it can reduce the time of obtaining a result by 4-5 times in the cases that sputum bacterioscopy yielded a positive result that allows the modified method to be applied. The advantages of the TB-Biochip technique are the time of detection multidrug-resistant MBT (24 hours), a possibility of obtaining these data just when analyzing sputum-isolated MBT DNA, and characterization of the MBT genomic elements that are responsible for drug sensitivity to antituberculous agents, by determining mutations in the examined genes and this all by using one chip. The agreement of results of microbiological and molecular genetic studies study of drug MBT sensitivity was 98%. There were no differences in the results of those using isoniazid. As for rifampicin, there was a difference in two samples (3.8%). Analysis of a combination of mutations forming multidrug resistance indicated that 74.3% of multidrug-resistant MBT isolates had mutations in the codon Ser531 > Lue of the rpoB gene and in the codon Ser315 > Thr of the katG gene. 97.4% of strains with signs of multidrug resistance had mutations in the codon 315 of the katG gene. 20.5% of isoniazid-resistant strains were observed to have mutations in two genes (katG and inhA) and 28.2% of the strains exhibited double mutation in the katG gene - Ser315Thr and Ile335 > Val.

[Organization of antituberculous care in penitentiary facilities]. / Organizatsiia pritovotuberkuleznoi pomoshchi uchrezhdeniiakh ugolovno-ispolnitel'noi sistemy.

The paper presents the organizational aspects of antituberculous care delivered at penitentiary institutions of the Russian Federation. The priorities of development, including the interaction with civil public health care of Russia and the continuation of tuberculosis-controlling activities jointly with international organizations, are identified.

[Tuberculosis in children: diagnosis of and methods for the evaluation of chemotherapy]. / Diagnostika i metod opredeleniia éffektivnosti khimioterapii tuberkuleza u detei.

The authors propose to determine altered forms of Mycobacteria, including sputum and blood granular forms, and the level of tuberculosis antibodies and serum Mycobacterium antigens by enzyme immunoassay as additional criteria for diagnosis of active primary tuberculosis in children. The diagnostic value of these indices is accounted for by the fact that they are found significantly more frequently and their level is significantly higher than the similar indices in healthy, tuberculosis-infected children and tuberculin-negative, prior BCG-vaccinated ones. By taking into account an decrease in the detection rate of sputum and blood granular mycobacteria and a change in the level of tuberculosis antibodies during chemotherapy, which are observed along with positive clinical and X-ray changes of a process, these tests should be recommended for evaluation of the efficiency of primary a bacillary tuberculosis in children.

[Improvement of diagnostics and methods of determining effectiveness of chemotherapy against tuberculosis in children]. / Sovershenstvovanie diagnostikik i metodov opredelneniia éffektivnosti khimioterapii tuberkuleza u detei.

Altered Mycobacterial forms, particularly granular forms, in the sputum and blood, as well as the serum level of tuberculosis antibodies and mycobacterial antigens by enzyme immunoassay are proposed to be determined as additional criteria for diagnosing active primary tuberculosis in children. The diagnostic value of these indices is accounted for by that they are more frequently detected and their levels are significantly higher than the similar ones in healthy, tuberculosis-infected children, and tuberculin-negative, earlier BCG vaccinated children. By taking into account a reduction in the detection rate of granular forms of Mycobacteria in the sputum and blood and altered levels of tuberculosis antibodies during chemotherapy, which were observed in parallel to the positive clinical and X-ray changes, these tests may be recommended for evaluation of the efficiency of treatment for primary abacillary tuberculosis in children.

[Use of native and lyophilized Loewenstein-Jensen's solid media in determining drug resistance on mycobacterium tuberculosis strains]. / Ispol'zovanie nativnoi i liofilizirovannoi sred Levenshteina-Iensena dlia opredeleniia lekarstvennoi ustoichivosti mycobacterium tuberculosis.

The results of determination of drug medium depend on both individual cultural properties of the strains tested and the type of the medium used. The composition of a medium is of significance for some strains: the latter grow in any medium and are characterized by the same spectrum of resistance. However, most drug-resistant strains have higher feeding demands which are not fully met on the lyophilized Löwenstein-Jensen medium. The strains in the lyophilized medium show an incomplete spectrum of resistance and less grow than do those in freshly prepared Löwenstein-Yersen medium.

[In vitro bactericidal action of dissolved ozone on different strains of Mycobacterium tuberculosis]. / Bakteritsidnoe vozdeistvie rastvorennogo ozona na razlichnye shtammy mikobakterii tuberkuleza in vitro.

The mycobacterium tuberculosis (MBT) strains H37 Rv (USA and Czechoslovakia), Erdman, and multiresistant clinical strain 2255 having different milliliter levels of microbes were kept in the soluble ozone at the concentrations of 1-4 micrograms/ml for 30 and 60 minutes. The soluble ozone-treated strains were cultured in the Finn-I medium. The soluble ozone at the concentration was found to produce bactericidal and bacteriostatic effects on MBT, including multiresistant strains. The MBT strains resistant to antituberculosis agents were also more resistant to soluble ozone; however, longer exposure to the "therapeutical" concentrations of soluble ozone proved to damage multiresistant strains.

[Biological properties of laboratory strains and clinical isolates of mycobacteria multiresistant to antitubercular preparations]. / Biologicheskie svoistva laboratornykh shtammov i klinicheskikh izoliatov mikobakterii, polirezistentnykh k protivotuberkuleznym preparatam.

Multiresistant M. tuberculosis strains show varying drug resistance, virulence and growth rates. The count of cells of some multiresistant strains in the guinea-pig parenchymatous organs after intracardiac inoculation was comparable with that of after inoculation with clinical isolates. Furthermore, some multiresistant strains were not inferior to sensitive clinical isolates. The findings lead to the conclusion that there is a wide range in the virulence of multiresistant strains and hence they can present an epidemiological hazard.

[Clinical presentation, course and treatment of recurrent sarcoidosis: followup data]. / Klinika, techenie i lechenie retsidiva sarkoidoza po dannym dispansernogo nabliudeniia.

The paper presents the results of long-term follow-ups of 107 patients with recurrent sarcoidosis registered as having Group VII-IB. In the patients, there was the greatest dissemination of the process in the lung, intrathoracic lymph nodes and other organs and MF MB in the blood, sputum, BAC? Group VIIIB patients need the greatest attention of physicians, treatment involving extracorporeal and physiotherapeutical methods to prevent respiratory failure and disability.

[Interaction of Mycobacteria isolated from sarcoidosis patients with antituberculosis antibodies]. / Vzaimodeístvie mikobakterií, bydelennykh ot bol'nykh sarkoidozom, s protivotuberkuleznymi antitelami.

In 80% of cases, antituberculosis antibodies from the sera of patients with tuberculosis were ascertained to react in enzyme immunoassay (EIA) with antigens (ultrasound disintegrants (USDs)) of reverse mycobacteria isolated (initially) from patients with sarcoidosis. The USDs of reverse mycobacteria isolated from patients with sarcoidosis reacted in EIA with monoclonal antibodies (MAb) against M. tuberculosis complex antigens (unique and crossover). Both common and distinctive (unique) antigenic determinants were detected via MAb against different mycobacterial types by immunoblotting in the antigenic complexes of M. tuberculosis H37Rv and reverse strains isolated from patients with sarcoidosis.

[Examining of humoral immunity on mycobacteria antigens in sarcoidosis]. / Izuchenie gumoral'nogo immuniteta na antigeny mikobakterii pri sarkoidoze.

Antibodies to Mycobacterium tuberculosis antigens H37Rv and reverse strains previously isolated from patients with sarcoidosis with granular isolates were determined in 50 patients with sarcoidosis (including 16 patients isolating granular types) and 56 patients with tuberculosis, by using ELISA and immunoblotting. Serum antibodies from patients with sarcoidosis were ascertained to more commonly react in ELISA with the antigen (ultrasound disintegrant (USDs) obtained from reverse mycobacteria isolated (initially) from patients with sarcoidosis (AGS) than with the USD of the M. tuberculosis H37Rv (AGT) and, on the contrary, serum antibodies from patients with tuberculosis more frequently reacted with the M. tuberculosis H37Rv. The spectrum of serum antibodies from patients with sarcoidosis greatly differed at immunoblotting with AGS and AGT. There was most commonly a reaction with the antigenic determinants 79, 27, 30, and 50 kDa to AGS and that of the determinants 17, 35, 32 kDa to AGT.

[Genomic polymorphism in Mycobacterium tuberculosis strains]. / Issledovanie genomnogo polimorfizma shtammov Mycobacterium tuberculosis.

Different genomic fingerprinting techniques (universal probes, such as rRNA genes, phage M13 DNA, IS 6110 probe) have been used to investigate the genomic polymorphism of Mycobacterium tuberculosis strains isolated in different geographical regions of Russia and in some CIS countries. As shown with the use of these techniques and a specially developed PCR-mediated system for genetic typing, M.tuberculosis strains are genotypically heterogeneous in regions with a sporadic level of tuberculosis morbidity and genotypically homogeneous in regions with elevated morbidity and mortality levels. The evaluation of the effectiveness of the genetic typing of M.tuberculosis with the use of different genomic fingerprinting techniques has made it possible to propose the optimum 3-stage scheme for the differentiation of M.tuberculosis strains: (1) the typing of all isolated strains of the PCR-mediated test system; (2) the typing of several selected M.tuberculosis strains with the use of 1S 6110 probe (2-3 strains of each detected PCR-RFLP [correction of PLRF] genotypes); (3) the typing of M.tuberculosis strains, containing 1 copy of 1S 6110 or not containing such sequence, with the use of probes (phage M13 DNA) detecting hypervariable sequences in M.tuberculosis genomes.

[Use of microscopy and inoculation for bacteriologic diagnosis in tuberculosis patients by the materials of Ivanovo region]. / Ispol'zovanie mikroskpii i poseva dlia diagnostiki bacteriovydeleniia u bol'nykh tuberkulezom po materialam Ivanovskoi oblasti.

The paper describes guidelines for isolation of Mycobacterium tuberculosis by bacterioscopy. It shows high detection rates for Mycobacteria in patients included into the programme. The detection rates of patients by bacterioscopy are not different in two quarters.

[New microbiological Techniques in diagnosis of tuberculosis]. / Primenenie novykh mikrobiologicheskikh tekhnologii v diagnostike tuberkuleza.

Microbial spectrum in patients with disorders of the upper respiratory tract consisted mainly of streptococci, staphylococci and pneumococci in 1991-1993, 1994 and later, respectively. Most specific for detection of nontuberculous flora was brushing with protecting agar plugs in combination with transport medium and anaerobic culturing. Direct immunofluorescence allows detection of antigens M. chlamydia in 29.4%, M. pneumoniae in 18.5% of cases. In diagnosis of tuberculosis the following new technologies were introduced: bacterioscopy with previous culturing of the material on liquid culture media followed by biological test and polymerase-chain reaction; determination of drug resistance based on nitrate reductase activity of M. tuberculosis indicating resistance of M. tuberculosis on liquid media in 4-7 days, dense egg media in 8-12 days, usage of Popesku medium enables correction of tuberculosis chemotherapy one month after the test initiation; M. tuberculosis identification using Western blot with monoclonal antibodies in some cases for 7 days.

[Advances in microbiological diagnosis of tuberculosis and sarcoidosis]. / Sovershenstvovanie mikrobiologicheskoi diagnostiki tuberkuleza i sarkoidoza.

Combined microbiological investigation of diagnostic material containing granular mycobacteria has revealed cultures-revertants both in tuberculous and sarcoidosis patients. Identification of the isolated cultures provided the conclusion on identity of granular mycobacterial structures discovered both in tuberculous and sarcoidosis patients.

[Detection of Mycobacterial antigens in tuberculous pleuritis, empyema and meningitis]. / Opredelenie mikobakterialnykh antigenov pri tuberkulëznykh plevrite, émpieme i meningite.

Mycobacterial antigens were identified by inhibition solid-phase enzyme immunoassay specific antibodies to M.H37Rv labelled with horse radish peroxidase in 20, 27, 24, 47 and 21 patients with tuberculous pleurisy, tuberculous empyema, meningitis, pleurisy and nontuberculous empyema, respectively. Mycobacterial antigens were found more frequently and in greater quantities in pleural and cerebrospinal fluids in tuberculosis than in the above nontuberculous affections. These differences were less pronounced in serum assays.